Open access peer-reviewed chapter

Value-Added Products from Natural Gas Using Fermentation Processes: Products from Natural Gas Using Fermentation Processes, Part 2

Written By

Maximilian Lackner, David Drew, Valentina Bychkova and Ildar Mustakhimov

Reviewed: 23 March 2022 Published: 19 May 2022

DOI: 10.5772/intechopen.104643

From the Edited Volume

Natural Gas - New Perspectives and Future Developments

Edited by Maryam Takht Ravanchi

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Abstract

Methanotrophic bacteria can use methane as their only energy and carbon source, and they can be deployed to manufacture a broad range of value-added materials, from single-cell protein (SCP) for feed and food applications over biopolymers, such as polyhydroxybutyrate (PHB), to value-added building blocks and chemicals. SCP can replace fish meal and soy for fish (aquacultures), chicken, and other feed applications, and also become a replacement for meat after suitable treatment, as a sustainable alternative protein. Polyhydroxyalkanoates (PHA) like PHB are a possible alternative to fossil-based thermoplastics. With ongoing and increasing pressure toward decarbonization in many industries, one can assume that natural gas consumption for combustion will decline. Methanotrophic upgrading of natural gas to valuable products is poised to become a very attractive option for owners of natural gas resources, regardless of whether they are connected to the gas grids. If all required protein, (bio) plastics, and chemicals were made from natural gas, only 7, 12, 16–32%, and in total only 35–51%, respectively, of the annual production volume would be required. Also, that volume of methane could be sourced from renewable resources. Scalability will be the decisive factor in the circular and biobased economy transition, and it is methanotrophic fermentation that can close that gap.

Keywords

  • methanotroph
  • biopolymers
  • polyhydroxyalkanoates (PHA)
  • polyhydroxybutyrate (PHB)
  • single-cell protein (SCP)
  • value-added chemicals
  • feed
  • food
  • scalability

1. Introduction

1.1 Methanol

Today, methanol is mainly produced from synthesis gas. It is a very versatile basic chemical. Processes to further process methanol include the following ones:

  • methanol to gasoline (MtG)

  • methanol-to-hydrocarbons (MtH)

  • methanol to olefins (MtO), and

  • methanol to propylene (MtP).

The most used strain to obtain methanol from methane by biochemical conversion is Methylosinus trichosporium. M. trichosporium was found to produce up to 4101 mg methanol/L/day [1]. Methanol is of interest, for example, for fuel cells and combustion, apart from being a basic chemical for synthesis.

In Ref. [2] immobilized Methylocystis bryophila were used to produce methanol by repeated batch fermentation from a simulated biogas mixture.

1.2 Biopolymers

Biopolymers are defined as either being biobased and/or biodegradable according to certain standards. They are used as thermoplastics, elastomers, and thermosets, to replace conventional polymers. Methanotrophs have been described to produce, for instance, PHB [1].

1.2.1 Polyhydroxyalkanoates (PHA) including PHB

The common energy storage compound in microorganisms is glycogen. When a shortage of essential nutrients occurs, particularly nitrogen or phosphorus, several microorganisms are able to store their energic carbon in a different compound, which is polyhydroxybutyrate (PHB). That PHB, which is accumulated intracellularly in granules up to 50% dry cell weight and above, can be extracted and used as a thermoplastic material. PHB is a biopolymer that is both biobased and biodegradable. The biodegradability can occur aerobically or anaerobically, in different environments. This property, together with the property set that is comparable to the commodity polyolefin PP (polypropylene), makes PHB an interesting bioplastics material. PHB belongs to the class of PHA (polyhydroxyalkanoates), which are polyesters and as such are naturally occurring compounds [3]. In plants, only PHB can be found [4], whereas microorganisms can produce a wide variety of PHA copolymers depending on available comonomers.

PHB formation was seen with type II methanotrophs. For instance, Methylocystis parvus OBBP was found to accumulate 60% of PHB on nitrogen limitation (ammonium), as opposed to only 36% accumulation with nitrate limitation. The strain Methylosinus trichosporium IMV3011 could accumulate 47% PHB with both nitrogen sources, nitrate and ammonium. Methylocystis hirsute could accumulate up to 51% PHB on ammonium (again after N-limitation). A total of 51% of PHB accumulation were found with a mixed culture dominated by Methylocystis GB25 on ammonium [1], see Table 1.

StrainPHB yield [g PHB/g CH4]Accumulation condition
Methylocystis GB250.4Sulfur deficiency
0.45Potassium deficiency
0.22Iron deficiency
0.52Ammonium deficiency
0.55Phosphorus deficiency
0.37Magnesium deficiency
Methylocystis parvus OBBP0.34Nitrogen limitation
Methylobacterium Organophilum CZ-20.43Nitrogen limitation
Mixed culture dominated by type II0.4Nitrogen limitation
Methylocystis sp. WRRC10.67Nitrogen and copper limitation

Table 1.

Yield of PHB found for selected strains of methanotrophs (type II).

Source: [1].

The yield that was obtained varied between 0.22 and 0.67 g of PHB per gram of methane.

EPS production can limit the yield. There were no extracellular products (EPS) under methane-limited growth conditions [5].

In Table 2, several bioreactor configurations to produce PHB were compared.

Bioreactor configurationpHTemp. [°C]CH4:O2 or airNitrogen source
5 L batch fermenter6.8–7.2301:3Nitrate
2 L batch fermenter7301:3Ammonium and nitrate
5 L fed-batch fermenter7341:1Ammonium and nitrate
70 L pressure bioreactors5.738pCH4 = 30%, pO2 = 15%Ammonium
1 L bubble column bioreactor7301:1Ammonium
2.5 L bubble column bioreactor with internal gas recirculation7.325Polluted air emission (4% CH4)
EBRT = 30 min
GRR = 0.5 m3/(m3 min)
Nitrate
1.4 L vertical loop bioreactor7301:1Ammonium
0.5 L jacketed stirred tank reactor7.225Gas flow 0.4 L/min
CH4 conc. 2 g/m3
Nitrate
4 L completely mixed batch reactor1:1Cyclic between ammonium and nitrate
4 L sequencing batch reactor30–321:4 (8 h)
1:1 (16h)
Nitrate
15.2 L fluidized bed reactor6.5–6.920–231:2.3Nitrogen gas

Table 2.

How different fermenters fare in PHB accumulation studies.

EBRT = empty bed residence time; GRR = gas recycling rate; Source: [1].

While PHB generally resembles PP in its property set, the material has a low elongation at break and due to its high crystallinity is brittle. Adding a few percent of valeric acid as a comonomer to PHB, yielding PHBV, makes the material softer and thereby more versatile. PHBV can be synthesized by methanotrophs, too. Another important PHA copolymer is PHBH, with a certain content of hexanoic acid being incorporated in the polymer.

Biopolymers can be an environmentally benign material class. For a life cycle assessment (LCA) for biopolymers made from biogas, see Ref. [6].

In Ref. [7], the yields were found to be 1.13 g of PHB per gram of methane for Methylosinus trichosporium OB3b and 0.88 g of PHB per gram of methane for Methylocystis parvus OBBP [7], which is significantly above the figures reported in Table 2, see Table 3.

ParameterUnitsModel value in LCA study [6]Observed for strain OB3bObserved for strain OBBP
(1)Percent PHB% [g PHB/g dry weight]502960
(2)Non-PHB biomass = [100/(1)] – 1g biomass1.002.450.66
(3)Methanotrophic growth yieldg biomass/g methane0.3450.630.73
(4)Methane requirement for the production of non-PHB biomass = (2)/(3)g methane2.903.890.92
(5)Yield of PHB on methaneg PHB/g methane0.551.130.88
(6)Methane requirement for the production of 1 g of PHB during PHB production phase = 1/(5)g methane1.820.881.13
(7)Total methane requirement for both phases = (4) + (6)g methane4.724.772.05

Table 3.

Production of PHB from methane.

The numbers are based on 1.0 g of PHB. Source: [8].

Methanotrophic strains producing PHB are listed in Table 4.

StrainCarbon sourcePHA structurePHA yieldMolecular weight
Methylocystis parvus OBBPMethanePHB0.34 g/g methane
Methylocystis parvus OBBPMethanePHB0.88 g/g methane
Methylosinus trichosporium OB3bMethanePHB18.1 mg/mg dry cell weight
Methylosinus trichosporium OB3bMethanePHB901.8 mg/L
Methylosinus trichosporium OB3bMethanePHB1.13 g/g methane
Methylosinus trichosporium IMV3011MethanePHB0.6 g/L1500 kDa
Methylocystis sp. GB 25 DSMZ 7674MethanePHB0.55 g/g methaneUp to 2500 kDa
Methylobacterium organophilum CZ-2MethanePHB233.3 mg/L
Methylocystis hirsutaMethanePHB and PHV0.63 g/g methane

Table 4.

PHB production by methanotrophs.

Source: [1].

Yield figures have a broad spread. PHB will form copolymers when comonomers, such as valeric acid, are supplied in the medium.

1.2.2 PHBV and other PHA

As stated above, polyhydroxyalkanoates (PHAs) are (bio)polyesters of hydroxy acids. They can be naturally synthesized by bacteria with the purpose of hoarding carbon under N- and P-limitation [9]. PHA consists of 3-, 4-, 5-, and 6-hydroxycarboxylic acids [10]. Lactic acid, citric acid, glycolic acid, malic acid, mandelic acid, and tartaric acid, by contrast, are alpha hydroxy acids (1-hydroxycarboxylic acids). Thereby, their polymers are no PHA. An example of a 2-hydroxycarboxylic acid (beta hydroxy acids, where the acid and hydroxy functional groups are separated by two carbon atoms) is salicylic acid. For hydroxybutyric acid, there are three isomers—alpha, beta, and gamma. When we talk about PHB, we typically mean poly(3-hydroxybutyric acid). Valeric acid (petanoic acid) has four isomers. It is found in some foods.

Polyhydroxyalkanoates (PHA) are established biopolymers. Some well-known brands and producers are given below in alphabetic order:

  • AirCarbon®/Newlight Technologies

  • EnMat/TianAn

  • Minerv-PHA™/Bio-On

  • Nodax™/Danimer Scientific

  • PHBH™/Kaneka

  • TephaFlex®/Tepha [11].

The organization Go!PHA [12] is promoting PHA.

To synthesize the copolymer PHBV, valeric acid is supplied as a comonomer to the fermentation broth.

It was found that above valerate concentrations of 0.7% (by volume), the PHBV accumulation in Methylocystis sp. WRRC1 was inhibited [1]. While the standard content of PHB, without any valerate present, was found to be 30%, it reached 15% only with high valeric acid concentrations. At 0.34% (by volume) of valerate, the PHBV content in the cells reached 60%, with 50% comonomer content [1]. Methylocystis parvus OBBP could be used to make different PHA from various supplied comonomers. 3-hydroxy-butyrate (3HB), butyrate, valerate, hexanoate, and octanoate were added and nitrogen limitation was applied [13]. The products included P(3HB-co-4HB), P(3HB-co-5HV-co-3HV), and P(3HB-co-6HHx-co-4HB). Table 5 shows PHBV production by methanotrophs.

MethanotrophsSubstrateCo-substratePHBV content [%]HV content [%]Biomass density [g/L]
Mixed culture dominated by Methylocystis sp.MethaneValerate 100 mg/L44201.2
MethaneValerate 400 mg/L30391
Methylocystis parvus OBBPMethanePropionate 100 mg/L3281.25
MethaneValerate 100 mg/L54221.82
MethanolValerate 100 mg/L5222
FormateValerate 100 mg/L5815
Methylosinus trichosporium OB3bMethaneValerate 100 mg/L50201.72
Methylocystis hirsutaBiogasValeric acid 130 mg/L54251.7
Methylocystis sp. WRRC1MethaneValerate 0.34%60503
MethaneValerate 0.34%, and biopolymer accumulation in a copper-free medium78583

Table 5.

PHBV accumulation from various substrates.

Source: [1].

Figure 1 shows how various PHA can be obtained by methanotrophs through suitable comonomer addition.

Figure 1.

Production of different PHA by comonomer choice. (a) Shows co-substrates without the hydroxy group and (b) With the hydroxyl group. Source: [13].

In Table 6, details on PHA yields from M. parvus OBBP with different comonomers are given.

Co-substrateswt% PHA polymerPHA monomer ratio [mol%]TSS [mg/L]
3HB3 HV4HB5HB6HHx
None42 ± 310000001600 ± 180
Butyrate (1.2 mM)55 ± 310000001660 ± 200
3-Hydroxybutyrate (1.2 mM)59 ± 510000001820 ± 220
4-Hydroxybutyrate (1.2 mM)50 ± 491.509.5001720 ± 240
Valerate (1.2 mM)54 ± 475.025.00001760 ± 160
5-Hydroxyvalerate (1.2 mM)48 ± 495.01.403.601640 ± 180
Hexanoate (1.2 mM)56 ± 410000001740 ± 200
6-Hydroxyhexanoate (1.2 mM)48 ± 397.601.001.41680 ± 220
Octanoate (1.2 mM)54 ± 310000001720 ± 180

Table 6.

Synthesis of different PHA through a variation of fatty acid co-substrates with the strain M. parvus OBBP.

TSS = total suspended solids. Source: [13].

Table 7 lists the physical properties of the obtained PHA.

PHA productsMolecular weightsThermal propertiesMechanical properties
MnMw/MnTm [°C]ΔHm [J/g]Tg [°C]E [GPa]σt [MPa]εb [%]
P3HB1.48E+061.821788333.043.25.2
P(3HB-co-24 mol% 3HV)1.32E+062.2414745−11.022.050.5
P(3HB-co-3.0 mol% 4HB)1.33E+062.1214865−21.235.6176
P(3HB-co-9.5 mol% 4HB)1.22E+062.0113547−50.831.2284
P(3HB-co-3.6 mol% 5HV-co-1.4 mol% 3HV)1.26E+062.1714444−20.829.9106
P(3HB-co-1.4 mol% 6HHx-co-1.0 mol% 4HB)1.27E+062.1115040−10.727.6134
Commercial P3HB7.38E+052.02
Commercial PHBV4.48E+052.18

Table 7.

Properties of the PHA materials derived by methanotrophic fermentation.

Mn = number average molecular weight; Mw = weight average molecular weight; Tm = melting temperature; ΔHm = apparent heat of fusion; Tg = glass transition temperature; E = Young’s modulus; σt = tensile strength; εb = elongation at break; Source: [13].

As Table 7 shows, the elongation at break is strongly increased by the comonomers, improving the biopolymer properties compared to heat PHB. Applications of PHA are described in Ref. [14].

1.2.3 Other biopolymers

Several microorganisms, also methanotrophs, produce EPS (extracellular polymeric substances, exopolysaccharides) [15, 16], which might be used potentially for biopolymer applications. A higher oxygen concentration increases the excretion of EPS by methanotrophs [8]. Methylobacter luteus, Methylomonas rubra, Methylococcus thermophilus, and Methylobacter ucrainicus were found to produce EPS from methane in the range of 0.5–0.8 g/L. In general, it is extreme conditions that promote EPS production. M. alcaliphilum 20Z, under moderately saline conditions, gave 1.8 g EPS/g of biomass when grown in a bubble column bioreactor for the treatment of dilute methane emissions [8].

A well-known biopolymer is PLA (polylactic acid). Its monomer, lactic acid, has been obtained from methanotrophs, too, see Table 8.

ProductStrainMethaneTemperatureCultivation typeProcess detailsTiter (productivity)
Lactic acidMethylomicrobium buryatense 5GB1S pLhldh mutant strain20% CH430°CBatchIncreased nitrate, phosphate, and trace elements in the medium0.808 g/L
Lactic acidMethylomicrobium alcaliphilum 20Z mutant strain20% CH4 (33% mock biogas in air)30°CContinuousBubble column bioreactor0.027 g/g DCW/h
Lactic acidM. buryatense 5GB1 mutant strain pAMR421% CH430°CBatchAmmonium as N source0.50 g/L
Crotonic acidM. buryatense 5GB1C mutant strain pCA0925% CH430°CBatch0.06 g/L
Butyric acid0.08 g/L
D-Lactic acidMethylomonas sp. DH-1, LA-tolerant strain JHM8020% CH430°CBatchIncreased nitrate1.19 g/L
Acetic acidMixed culture dominated by Candidatus “Methanoperedens nitroreducens”90% CH4Room temperatureBatchNitrogen limitation0.097 g/L (1620 μmol/L)
Muconic acidM. buryatense 5GB1 mutant strain pMUC20% CH430°CSemi-continuousCSTR continuous gas supply0.012 g/L
Succinic acidMethylomonas sp. DH-1 mutant DS-GL strain30% CH430°CBatchpH 6.90.195 g/L
3-HP acid (hydroxypropionic)Methylosinus trichosporium OB3b mutant MCRMP strain30% CH430°CBatchpH 7.00.061 g/L
4-HB acid (hydroxybutyric)M. trichosporium OB3b 4HB-SY4 mutant strain40% CH430°CBatch0.011 g/L (10.5 mg/L)

Table 8.

Organic acids from methanotrophs.

The table gives the titer (g/L). Source: [17].

PLA is biobased and degradable (however, it requires 70°C for disintegration, so “home compost” standards are not met by PLA-based bioplastics products; The material is only “industrially compostable,” see EN 13432 standard). PLA is suitable for food packaging (FDA rating as gras = generally recognized as safe). Today, PLA is made from sugar-derived LA, which requires agricultural starch or sugar production with the associated food/feed competition over land, fertilizer and water consumption, etc. Significant efforts are undertaken to make the enzymatic hydrolysis of lignocellulosic biomass economically viable, however, no breakthrough has been achieved in that field yet (compare 2nd generation biofuel production attempts). Methane-derived PLA can offer a lower environmental footprint and be produced with less price volatility than agriculture-based material. Of PLA, several copolymers and blends exist, e.g. with glycolic acid (PLA + GLA, PLGA), as well as compounds. For LA production by methanotrophs, see [17, 18].

1.3 Other value-added products accessible through methane fermentation

Using natural and engineered strains, not only protein and biopolymers can be obtained from methane, but also several other compounds, for example

  • Isoprene [19]

  • Squalene [20]

  • Succinic acid [21]

  • Methanobactin [22]

  • Ectoine [23, 24]

  • Muconic acid [25]

  • Astaxanthin [26]

For a recent review on bioproducts from methane using methanotrophs, see Ref. [17]. A methanotroph-based 2nd generation biorefinery, that is, single-input, multiple-output configuration is treated in Refs. [23, 27]. In general, multi-product biorefineries are better from an economical point of view for methane-based industrial biomanufacturing. For biorefinery downstream processes, see Refs. [28, 29, 30].

1.4 Protein from methane: Single-cell protein (SCP)

Protein is a vital part of our diet. It can be found in plants and meat. For several decades, there has been interest in single-cell protein (SCP), that is, protein derived from yeasts, algae, and bacteria [31, 32]. Yeast cells have been used to make beverages and bread for over 4500 years [33], and also algae have a long history of food. Bacterial SCP is of particularly high interest for commercial production as will be outlined below.

There exit proteinaceous methanotrophs, which contain high amounts of protein made from methane. Bacterial SCP, also called BPM (bacterial protein meal), can be obtained from methane with methanotrophs, in pure and mixed cultures, see Refs. [34]. Continuous aerobic fermentation [35, 36] is generally preferred for higher yield.

A mixed culture with Methylococcus capsulatus (Bath), complemented by Ralstonia sp, Brevibacillus agri, and Aneurinibacillus sp. [37] was described to yield SCP. Also, SCP production from methanol has been reported [38], using methylotrophs.

Applications of SCP are mainly in the feed and food sector. In 1997, the United Nations “Protein-Calorie Advisory Group” discussed the safety of SCP for Animal and Human Feeding [39], apart from other commissions and panels [40, 41, 42].

Twenty-three years earlier, in November 1974, the European Association of Single-Cell Protein Producers (Association européenne des producteurs de protéines unicellulaires) UNICELPE had been founded [43].

SCP produced on a commercial scale has been reported to be deployed in these areas:

  • As animal feed, for example, for poultry, calves, mink, and pigs.

  • As food additives, for example, vitamin carrier, aroma carrier, and emulsifying agent.

  • In industrial processes, for example, as foam-stabilizing agents and in the processing of leather and paper [33, 44, 45].

For food, another application is to boost the nutritional value, for example, of baked food items, ready-made meals, and soups. Single cells have also been used in the food industry as starter cultures (e.g., in bread, beer, and wine making by various yeasts) [33, 44, 45].

SCP has been described as antiobesity food [46]. A patent [47], EP 79.1641475, discloses the use of lipids from methanotrophs for cholesterol reduction. The use of biogas as feedstock for methanotrophs is detailed in Refs. [48, 49]. In Ref. [50], the simultaneous production and use of SCP and PHB bioplastics are discussed.

1.4.1 History of SCP

In Germany during World War I and II, yeast was used to produce food for soldiers, prisoners, and the civilian population [51, 52]. Also, Russia and Japan used similar single-cell proteins for food [51]. Different feedstocks had been tested and used, also fossil-based ones, which gave a negative touch to the products in some peoples’ eyes. Hence, in the year 1966, the term “single-cell protein” (SCP) was introduced by Carl L. Wilson, building upon prior successful productions of protein from fossil sources, to provide a more positive term than “petroprotein.” In fact, in the Soviet Union, early attempts were made to obtain cost-effective protein from oil. “BVK” (belkovo-vitaminny kontsentrat = protein-vitamin concentrate) plants close to oil refineries in Kstovo (year 1973) and Kirishi (year 1974) were built [33]. Till 1989, eight such factories had been constructed by the Soviet Ministry of Microbiological Industry [33].

In Russia, the single-cell protein variants were called “Gaprin” (SCP from methane) [53], “Paprin” (SCP from paraffins), [54, 55] “Meprin” (SCP from methanol), and Eprin (SCP from ethanol). The total capacity was estimated at 1.5 million tons of SCP per year [56].

Work was pioneered by Alfred Champagnat [57, 58].

For petroleum-derived SCP, see [59, 60, 61, 62].

Also in Western Europe, similar attempts were made. British ICI (Imperial Chemical Industries Ltd) succeeded in commercializing Pruteen™ single-cell protein. As feedstock, methanol was used, deploying Methylophilus methylotrophus in a 1500 m3 airlift fermenter [63, 64]. That production was a major milestone in biotechnology [65]. It can be considered the foundation for mycoprotein Quorn™ [66, 67], which is made from a fungus (Fusarium venenatum) from sugar and sold on the order of several 10,000 tons/year today as a meat replacement.

The historic Pruteen™ plant was designed for an output of 50,000–75,000 metric tons/year. The investment was $70 million (1979), and development costs had been $20 million [68].

Scientific American [69] wrote in 1981: “Beer, Wine, bread, and cheese have been made by microorganisms since Neolithic times. To them have been added spirits, yogurt, pickles, sauerkraut, Oriental fermented foods, and today single-cell protein.”

The BP (British Petrol) SCP process for “Toprina” is described in Ref. [70].

French activities from SCP from methane are reviewed in Ref. [71].

German Hoechst/Uhde had developed Probion™ as a single-cell protein from Methylomonas clarae with 70% protein content [56]. A purified version, with 90% protein content with the nucleic acids and fats being removed, was intended for human consumption under the name Probion-S [56].

Liquipron™ (by Liquichimica from Italy) production is detailed in Ref. [72].

Another early, large single-cell protein project was Bioprotein™, which company Norferm (owned 50–50 by Statoil and Anglo-Norwegian pharmaceuticals group Nycomed Amersham [73], later DuPont) developed. Out of Norferm, the companies Calysta and UniBio developed. The Norferm SCP plant can be seen in Figure 2. Bioprotein™ was made from methane.

Figure 2.

The historic Norferm single-cell protein production facility at Tjeldbergodden, Norway. Source: [74].

In the Norferm process, methane (from natural gas), oxygen, ammonia, and minerals were fermented. The concentration of the biomass was 20 g/L, and the temperature was 45°C in a continuous process. By centrifugation, the biomass was concentrated to 80–90 g/L in centrifuges and then further to 220 g/L by ultra-filtration. By a “short intensive heat process, the cells are partially opened to improve the digestibility of the protein.” Spray drying is used to dry the biomass. The specific growth rate was 0.2/h, corresponding to biomass productivity of 4 kg/h/m3 [74].

These numbers are confirmed from additional sources; industrial production of SCP is feasible at an output of 4.0 kg biomass/m3 reactor/h with an expected yield of 0.8 g biomass/g CH4 [75]. The process is shown in Figure 3.

Figure 3.

Process diagram of the single-cell protein process at Norferm. Source: [74].

The cooling requirement amounted to 62.3 MJ/kg of biomass, and the total volume of the loop reactor measured 300 m3. As a strain, M. capsulatus was used to yield the following product composition:

  • Crude protein: 70%

  • Fat: 10%

  • Fiber: 1%

  • Carbohydrates: 12%

  • Minerals: 1%

Norferm’s BioProtein® (BP) SCP had been approved in the EU “as a protein source in animal feeds since 1995, for fattening pigs (8%), calves (8%) and salmon (19-33%)” [76].

Table 9 provides an overview of several other commercially implemented SCP processes from the “golden age” of the industry around the year 1977 when also licensing options were available [78, 79]. An interesting retrospective view is provided by [80].

OrganizationLocationSubstrateOrganismProduct nameCapacity [tons/year]Status
British Petroleum (BP)Lavéra, FranceGas oilCandida tropicalisToprina16,000Shut down
British Petroleum (BP)Grangemouth, Scotlandn-ParaffinsCandida lipolyticaToprina4000Operating
BP-ANICSarroch, Sardinian-ParaffinsCandida lipolyticaToprina100,000Constructed, awaiting government approvals
Liquichimica (with Kanegafuchi)Saline di Montebello, Italyn-ParaffinsCandida maltosaLiquipron100,000Constructed, awaiting government approvals
USSRSeveral sitesn-ParaffinsCandida spp.BVK300,000 or moreOperating
Imperial Chemical Industries (ICI)Teesside, UKMethanolMethylophilus methylotrophaPruteen50–75,000Construction approved, start-up late 1979
AmocoHutchinson, Minnesota, USAEthanolTortula yeast (Candida utilis)Torutein5–7000Operating

Table 9.

Commercial-scale SCP processes with hydrocarbons and alcohols and feedstocks.

Source: [77].

Another well-known SCP derived from n-alkanes (paraffins) was Fermosin™ [61].

Table 10 gives additional projects (status 1977).

OrganizationLocationSubstrateOrganismStatus and capacity [tons/year]
Societé Nationale Sempac: Engineering by Process Engineering Co. (PEC; Member of Chemap, Switzerland)AlgeriaMolassesYeast20,000Project start December 1977 (Algiers)
BulgariaMethane, methanol, n-paraffinsDepends upon the process chosen100,000Discussions with ICI, BP, Shell, USSR, location will depend on the process chosen
Alberta Gas Chemicals with Alberta Research Council, Celanese Canada, with Mitsubishi Gas ChemicalCanadaMethanol, n-paraffinsYeast100,000Study, for a plant in Medicine Hat (Alberta/Canada)
CzechoslovakiaEthanolYeast4000 (pilot plant); in planning: 100,000 t/a in Kojetin/Northern Moravia; pilot plant in operation
Joint East Germany/Polish project using USSR/East German-developed processEast GermanyGas oilYeast60,000 t/a in Schwedt
Groupement Francais des Proteines (50% owned by Institute Francais du Petrole; other 50% shared by Elf-Erap and Cie-Francaise des Petroles)Francen-ParaffinsYeastPilot plant (Soleize); earlier plans for 100,000 tons/year plant canceled
Institute of PetroleumIndia“crude oil”YeastPilot plant (Gujarat, near Baroda)
Hebrew University (Yissium Research Devel. Co.) with Dor Chemicals (Haifa)IsraelMethanolBacteria50 tons/year pilot plant to be scaled up to 1000 and then to 25–100,000 tons/year
MontedisonItalyEthanol; carbohydrateDepends upon the process chosenTwo small pilot plants. Ethanol jointly with Czechoslovakia, carbohydrate jointly with PEC, Switzerland; joint marketing study with Amoco
Societa Italiana Resine S.p.A., Milan (SIR)MethanolCandida boidiniPilot plant (Milan)
Dainippon Ink and Chemicals (with Koa Oil)Japann-ParaffinsYeastFoundation laid for 60–120,000 tons/year plant; canceled
Kanegafuchi (with Maruzen)n-ParaffinsYeast60,000 tons/year plant; canceled
Mitsubishi Ga Chemical Co.MethanolYeast, bacteria4000–5000 semicommercial (Niigata)
Mitsubishi PetrochemicalEthanolYeast
Kyowa Hakko Kogyon-ParaffinsYeast100,000 tons/year; canceled
Mitsui Toatsu Chemicaln-ParaffinsYeast50–60,000 tons/year; canceled
Asahi Chemicaln-ParaffinsYeast50–60,000 tons/year; canceled
Sosa Texcoco SAMexicoCO2Spirulina maxima1 ton/day (Mexico City)
Roniprot L.L.A.; Technology from Japan (reports of negotiations with Ron, Dainippan, Sumito Shoji Kaisha)Romanian-ParaffinsYeast60,000 tons/year (Arges; Jassyon) said to have started construction in 1973
Petromin (state-owned Saudi Arabian Oil Co.) with British PetroleumSaudi Arabian-ParaffinsYeastStudy for 100,000 tons/year (Al Jubail)
Instituto de Fermentaciones Industriales, Centro Superior de Investigaciones Cientificas (government sponsored, but autonomous)SpainEthanolHansenula anomalaPilot plant, 100 kg/day (Madrid)
Norsk Hydro and AB Marabou formed “Norprotein” (financial support from Nordic Industrial Fund and Swedish Council for Technical Development)SwedenMethanolPilot plant planned (Sundbyberg)
Chi Yee Solvent Works, Chinese Petroleum Corp.TaiwanKerosene, fuel oil, gas oilPseudomonas
Phillips PetroleumUnited StatesMethanolBacteriaPilot plant, Bartlesville, Oklahoma
Bio Proteinas de Venezuela (Venproteinas) (with BP), Stone and Webster, contractorVenezuelan-ParaffinsYeast100,000 tons/year (early 1979) (Puerto la Cruz)
Schick-Chemie-Technik GmbH (Cologne), with technology licensed from the Institute of Industrial Fermentation, Spain (see above)West GermanyEthanolHansenula anomala100,000 tons/year (several 50 m3 fermenters) planned for 1978
Society for Biotechnological Research (Braunschweig Stoeckheim) with Wintershall AG (subsidiary of BASF)“Oil”Aerobic mycobacteriaEffluent to be used for secondary oil recovery in a field test in Lower Saxony
Friedrick Uhde GmbH; Hoechst AG (Gelsenberg has withdrawn), with government supportMethanol (also n-paraffins)Methylomonas sp., Candida lipolyticaPilot plant planned (earlier plans for demonstration plant (100 tons/year) at Gelsenkirchen-Horst canceled), ultimate plans for 100,000 tons/year
Kohlenstoffbiologische Forschungsstation, E.V., DortmundCO2Scenedesmus acutus200 tons/year

Table 10.

Additional SCP projects as of 1977.

Source: [77].

The overview above gives testimony of a fairly developed industry, which then disappeared again from the market. Fifty years ago, SCP succeeded technologically, but then failed economically, mainly due to cheap soy prices and increasing fossil fuel costs, as this quotation for the pertinent literature exemplifies:

“By 1974, Shell announced plans for enlarged pilot plant facilities at Sittingbourne, and a development program in Amsterdam, with the ultimate goal of constructing a 100,000 ton/year plant. In the spring of 1976, however, Shell announced that it had stopped work on commercialization, and the expansion and development plans were “indefinitely postponed.” The decision was said to be based on three factors—the low price of soybeans and maize; the potential in large areas of the world for expanding existing sources of protein; and the difficulty of applying Shell’s sophisticated process in “underdeveloped countries” [77].

So Shell bailed out of SCP within a short period of time, after prior heavy investments. That step cannot be considered unusual, compare to the press release from Dow in 2005 [81], when Dow stepped out of the bioplastics business “due to slow sector maturation” after having invested an estimated 750 million USD.

Four seminal books on SCP are [51, 82, 83, 84].

1.4.2 Use of SCP: Feed and food

SCP has multiple possible applications, both in the feed sector and in the food sector [33, 48, 85], see Table 11:

Potential application for SCP in animal diets (feed)Potential application for SCP in foodstuff
• “Calves having fattening ability.
• Poultry having fattening ability.
• Pigs having fattening ability.
• Fish breeding.
•Laying hens feed.
• Feeding of household animal” [65, 86]
• “As carriers of vitamin.
• As emulsifying agent.
• As carriers of scent.
• In soups.
• Improving the nutritional worth of baked items.
• In readymade meals.
• Within food recipes” [65, 86]

Table 11.

Where SCP could be used.

Source: [33, 48].

Kuzniar et al. have suggested the use of methanotrophic bacterial biomass as a mineral feed ingredient for animals [87].

Also, applications in paper processing, leather processing, and foam stabilization are mentioned in the literature [33, 48].

In Ref. [88], SCP is discussed as a basis for microbial growth media, and in Ref. [89] for wood adhesives.

For (animal) feed applications, the dried bacterial biomass, which typically contains 70% of protein, can be fed directly. In SCP for (human) food applications, the content of nucleic acids (NA) has to be reduced from 10–15 to approximately 1–2%, which can be done thermally or enzymatically. Nucleic acid reduction by heat shocks is described in Ref. [90].

1.4.3 Properties of bacterial SCP

Table 12 summarizes the properties of different SCPs:

FeatureBacteriaYeastFilamentous fungiAlgae
Growth rateHighestQuite highLower than bacteria and yeastLow
SubstrateA wide range of substratesMost substrates except hydrocarbons and CO2Limited substrates (mostly starchy and cellulosic materials)Light, inorganic carbon sources, for example, CO2 (preferably)
pH range5–75–73–8up to 2
Cultivation systemBioreactorsBioreactorsBioreactorsOpen ponds, tanks in sunlight
Risk of contaminationHigh; precautions are necessaryLowLow if grown below pH 5High
Biomass recoverySometimes problematic; new improved methods are neededEasy by centrifugationEasy for filamentous or pellet formsDifficult and costly with unicellular algae
Protein content (crude)80% or more55–60%50–55%Up to 60%
Amino acid profileGenerally good, a small deficit in S-containing acidsGenerally good, deficit in S-containing amino-acidsLow in S-containing amino acidsGenerally good; low in S-containing amino acids
Nucleic acid contentHigh (8–14%)High (5–12%)High (3–10%)Low (4–6%)
Removal of nucleic acidsNecessary
ToxinsGram-negative bacteria may produce endotoxinsMany species produce mycotoxinsThree types of toxins: endotoxin, neurotoxins, heptotoxins
Other featuresHigh vitamin B contentChitin may contain a significant proportion of N content, which is unavailableLow yield (1–2 g dry wt/L). High chlorophyll content is unsuitable for humans

Table 12.

Comparison of some cultural and biochemical characteristics of various microbe groups to make SCP.

Source: [91].

Bacteria are very well suited to make SCP, see Table 5. They exhibit the highest growth rates. On the other hand, the nucleic acid content is elevated compared to SCP by other microorganisms. The extraordinary growth performance of bacteria is also visible in Table 13.

OrganismMass doubling time
Bacteria10–120 min
Algae/molds2–6 h
Yeasts10–120 min
Plants1–2 weeks
Chickens2–4 weeks
Pigs4–6 weeks

Table 13.

Mass doubling time of different organisms.

Source: [91].

With doubling times of the mass on the order of 10–100 minutes, bacteria and yeasts grow incomparably faster than plants or animals. This translates into unsurpassed productivity, as Table 14 illustrates.

Organism (1000 kg)Amount of protein produced in 24 h [kg]
Beef cattle10−1
Soybeans101
Yeast102
Bacteria1012

Table 14.

Comparison of protein production efficiency of selected organisms over 24 h.

Source: [91].

Within 24 h, a starting mass of bacteria of 1000 kg can yield, theoretically, 1012 kg of protein, whereas beef would only produce 0.1 kg and soy in the order of 10 kg. An analysis of bacterial meal (BM) derived from methane is given in Table 15.

FeedstockBacterial cultureCrude proteinLipidsAshNucleic acids
MethanolMethylophilus methylotrophus81.37.29.115.9
MethaneMethylococcus capsulatus, Ralstonia sp., Brevibacillus agri, Aneurinibacillus sp.73.210.78.59.9
68.110.48.0Not determined
68.78.08.0Not determined
73.48.47.7Not determined
71.98.36.7Not determined
69.58.16.211.1
67.09.96.4Not determined

Table 15.

Properties of BM (bacterial meal) made from methane and methanol, based on g/100g of dry mass.

Source: [65].

As one can infer from Table 15, the protein content approaches and exceeds 70%. Table 16 shows the protein content of SCP from selected microorganisms.

BacteriaSubstrateSCP (%)Reference
Haloarcula sp. IRU1Petrochemical wastewater76[92]
Methylococcus capsulatus, Ralstonia sp., Brevibacillus agriMethane (natural gas)67–73[65]
Aneurinibacillus sp., Methylomonas and Methylophilus spp., Metilococ capsulatusGas and liquid products of sewage<41[93]
Methylococcus capsulatus (bath)Methane53[94]
Methylophilus spSupernatant and biogas24[93]
Methylocapsa acidiphilaMethane59[95]
Methilomonas sp.Natural gas69.3[96]
Methilomonas sp.Biogas and supernatant of sewage sludge56[93]

Table 16.

Protein content of bacteria expressed as SCP (single-cell protein) grown on different substrates.

Source: Excerpt from [44].

Another important aspect of SCP is its quality, which can be expressed by the amino acid profile. In Table 17, the composition of amino acids of bacterial SCP compared to other proteins is shown.

Amino acidsBMSoybean mealFishmealMethanol-grown bacterial protein (Pruteen™)
Indispensible amino acidsArginine6.37.46.24.6
Histidine2.22.72.51.9
Isoleucine4.44.74.74.3
Leucine7.57.57.97.0
Lysine5.66.18.26.0
Methionine2.61.33.02.4
Phenylalanine4.25.04.14.1
Threonine4.33.94.04.6
Tryptophan2.21.40.90.9
Valine5.84.85.35.6
Dispensible amino acidsAlanine7.14.26.17.1
Apartic acid8.511.29.98.8
Cysteine + cystine0.71.50.90.7
Glutamic acid10.618.212.610.6
Glycine4.94.26.05.7
Proline3.85.04.32.9
Serine3.65.24.13.3
Tyrosine3.63.83.23.4

Table 17.

Amino acid profile of SCP made from natural gas (BM = bacterial meal).

For reference, the data for soybean meal and fishmeal are provided. The rightmost SCP was obtained from methanol, Pruteen™, by Methylophilus methylotrophus [g/16 g N ± standard deviation of free, hydrated amino acids]. Source: [65].

Bacterial SCP is approved as a feed ingredient in the EU feed catalog (EU 68/2013) [63]. SCP for feed has been tested extensively, see Table 18 for an overview.

SCP-protein evaluations by animal tests
OrganismDigestabilityProtein efficiency ratioBiological value
Algae65–860.7–2.648–81
Bacteria80–9073–82
Fungi10–75
Yeast81–960.9–1.732–69
Yeast and methionine962.0–2.391–96
FAO/WHO reference protein100100

Table 18.

As early as 1981, SCP was tested for feed.

Source: [97].

SCP feed trials were made with Drosophila [87] several monogastric species [98], including rats [99, 100], pigs [61, 100, 101, 102], dogs [45, 103], (lactating) cows [104], veal calves [105], chickens (broilers) [72], mink (Mustela vison), fox (Alopex lagopus), Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss) [106], Pacific White Shrimp (Penaeus vannamei) [107, 108], Atlantic halibut (Hippoglossus hippoglossus), tilapia (Oreochromis niloticus, Oreochromis mossambicus) [109, 110], Japanese yellowtail (Seriola quinqueradiata) [111], zebrafish (Danio rerio) [112] and other aquaculture species [113, 114]. In feed, SCP can replace fish meal and soy. It was found in Ref. [65] that “bacterial meal (BM) derived from natural gas fermentation, utilizing a bacteria culture containing mainly the methanotroph Methylococcus capsulatus (Bath), is a promising source of protein based on criteria, such as amino acid composition, digestibility, and animal performance and health.” [65].

The oral immunogenicity of bacterial SCP was tested by [115].

The use of SCP for feed and food was also proposed by John H. Litchfield in 1979 [116].

An example from Germany (1943–1949) is the “wood sausage,” a spread made in Wildshausen from paper production waste (sugars) using the fungus Oidium laktis. The process was licensed from Biosyn GmbH [117]. Other yeast spreads on the market are Marmite, Cenovis, and Vegemite: “Spent brewer’s yeast (Saccharomyces cerevisiae) have been sold for more than a century in yeast extracts such as Marmite® (Unilever and Sanitarium Health Food), Vegemite® (Bega Cheese Ltd.), Cenovis® (Gustav Gerig AG), and Vitam-R® (VITAM Hefe-Produkt GmbH).” [117]

“Another commercially available yeast, Torula (Candida utilis, renamed as Pichia jadinii), a widely used flavoring agent, is also high in protein. Torula was used in Provesteen® T, produced by the Provesta Corporation in the 1980s, along with similar products using Pichia and Kluyveromyces yeast)” [118].

1.4.4 Aquatic species

Approximately half of the global fish and aquatic species production comes from aquaculture operations, with an increasing share. Fish meal is volatile in price and, due to overfishing, is not a sustainable feed material. The same holds true for soy; while soy is cheap, its immense monoculture production has resulted in rainforest destruction on a global level. It was shown that soybean meal-induced enteritis in Atlantic salmon (Salmo salar) can be prevented by cell wall fractions obtained from Methylococcus capsulatus [119].

1.4.5 Chicken

Various tests were carried out to feed chickens with bacterial SCP. SCP could maintain chicken growth performance and increase the feed to gain (feed conversion) and reduce the fat content in chicken [120]. “Replacing soybean meal and small amounts of rendered fat with BPM caused less intensity of odor and less rancid flavor of chicken meat” [120].

Processed animal protein (PAP) from poultry/pig for poultry/pig [121] is currently being investigated, too, but brings back memories about BSE [122]. Also, potential quantities are limited.

1.4.6 Mink

Another species where SCP was tested successfully is mink (Mustela vison). SCP digestibility tests were carried out in 2009 by the Department of Animal and Aquacultural Sciences at The Norwegian University of Life Sciences, Ås [123]. Mink are of importance for the fur industry.

1.5 Status of single-cell protein as food

Oral delivery of bacteria is not novel [124]; for an overview of beneficial microorganisms in food and nutraceuticals, see Ref. [125]. Microorganisms in food are covered in Refs. [126, 127, 128].

In the literature, there is ample coverage of food trials of SCP in the last 50 years.

One example is a trial from 1979 to include SCP “Pekilo” in sausages and meat balls [129].

SCP has a peculiar taste. Some variants, which have a high content of glutamic acid, have been proposed and used instead of monosodium glutamate (MSG) [118] as the flavor enhancer in food.

Pekilo is a microfungus-derived SCP from Paecilomyces varioti based on pulp and paper waste streams. It is now being revived by Enifer [130]. Another work with SCP from waste is discussed in Ref. [131], SCP with the fungus P. chrysosporium in [132].

As early as 1971, single-cell protein had been subjected to clinical testing in tolerance trials in adults [133]. For immunogenicity testing of food proteins, see Ref. [134].

For humans, it is recommended that the daily nucleic acid (NA) intake does not exceed 2 g, as higher quantities (particularly RNA and, to a lesser extent, DNA) have been reported to increase the content of uric acid in blood plasma to an unhealthy level [135], leading to gout [136]. As SCP contains a comparatively large amount of nucleic acids, it needs special processing for food applications. For feed, by contrast, the nucleic acid species do not harm. In Table 19, approaches for RNA reduction in SCP are summarized.

MethodProtocolOrganismInitial and final % RNA (of dry wt)
Base-catalyzed hydrolysis0.12 N NaOH for 30 min at 50°CP. variotii9%→3%
Chemical extraction5% NaCl at 120°CBaker’s yeastComplete removal
Cell disruption; physical separation:Disintegration-glass beadsDifferent microorganismsFinal % RNA = 2
(a) Enzymatic treatmentMalt sprout nucleaseYeastFinal % RNA < 3
(b) ChemicalSuccinic anhydride, pH 4.2–4.4S. carlsbergensis6–8%→1.8%
(c) ChromatographyCellex E-Ecteola celluloseC. utilis, S. cerevisiae, S. carlsbergensis, Z. mobilisLess than 3% of initial content
Exogenous RNAaseBovine pancreatic ribonuclease, 55–56°C, pH 6.7–8.0S. utilis, C. intermediaFinal % RNA = 1.5
Endogenous RNAase90°C, pH 2, 20 minR. glutinis6.5%→1.2%
-heat shock45°C, pH 5.8–6.9, 2 hC. utilis6–8%→2%
68°C for a few sec, 50°C, 1 h, 55°C, 1 hC. utilis7–8%→1–2%
50–60°C + NaClS. cerevisiae7–8%→1.4%

Table 19.

RNA reduction in SCP.

Source: [51].

Tables 2023 compare different treatments of SCP to reduce nucleic acids [137].

Amino acidsYeast cells [mg/g]Protein extracted with NaOH [mg/g]
Cystine + cysteine8.1511.88
Lysine + histidine10.8116.55
Aspargagine7.5815.5
Aspartic acid1.893.54
Serine + glycine7.828.86
Glutamic acid4.115.72
Threonine26.1133.1
Alanine29.9432.79
Tyrosine11.8914.77
Methionine + tryptophan6.9411.96
Phenylalanine7.149.45
Leucine + isoleucine22.8537.28

Table 20.

Change of the amino acid composition of yeast cells by NaOH extraction.

Source: [137].

NaCl [%]RNAReduction [%]
14.546.4
24.0516.5
33.6824.1
43.1934.2
54.0516.5
Control4.850.0

Table 21.

Reduction of RNA in yeast cells (Candida lipolytica YB-423) by NaCl treatment, incubation at 50°C for 20 min.

Source: [137].

Type of proteinTotal nitrogen [%]Total protein [%]RNA [%]DNA [%]
Yeast cells6.7242.004.902.13
Protein extract10.8067.501.220.399

Table 22.

Nucleic acid and protein content of yeast cells by extraction with NaOH.

Source: [137].

Temperature [°C]Time [s]RNA [%]DNA [%]Reduction of nucleic acids [%]
6053.3251.46031.93
103-1501.37535.63
202.7121.35042.22
402.6251.32043.88
802.4501.30050.92
7052.1871.25051.11
101.6621.24058.72
201.5751.23060.10
401.4001.20063.02
801.0001.15069.42
Untreated cells4.9002.1300.0

Table 23.

RNA and DNA in yeast cells (C. lipolytica YB-423) treated by heat shock.

Source: [137].

There is currently a strong movement toward alternative protein (alt protein), as concerns over the healthiness and sustainability of meat, first and foremost imported beef, are increasing all over the world. Hence there is good potential for SCP for food applications. By the end of 2021, Solein™ submitted its dossier for SCP food approval in the EU [138].

Occupational health aspects of SCP are discussed in Ref. [139], where it was found that inhalable dust needs to be avoided.

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2. Operational aspects of methane fermentation

Methane fermentation occurs naturally and is carried out in bioreactors. Medium optimization and mineral requirement are treated in Refs. [140, 141, 142].

The reactions can be described as follows for methane limitation 4.9 g is for bacteria and products together [5]:

CH416g+1.8O256gBacteria16.1gdrywt.+0.34CO215.8gE1

Under oxygen limitation, it was found [5]:

CH416g+0.3O29.6gBacteria4.9gdrywt.+Products+0.05CO22.2gE2

By contrast, [96] found for methane limitation:

4.25gO2+1.245gCH41gcells+2.0gCO2E3

For oxygen limitation, this formula could be determined [72]:

2.5gO2+1.01gCH41gcells+1.216gCO2E4

For fermentation technology in general, see Refs. [143, 144, 145, 146], for its economics [147].

2.1 SCP production process

As SCP has already been produced at a large scale some 50 years ago, see above, there is ample experience with gas fermentation. SCP production is summarized in Table 24.

Cultivation operationGrowth modalityCapital and operational considerationsEmerging commercial examples
Aerobic bioreactorHeterotrophs
Mixotrophs
High cell mass yield
High capital costs
High energy consumption
Sterile operation
Significant installed industrial capacity
Methanol, glycerol, or ethanol—KnipBio
Glucose—Veramis
Cellulose—Arbiom, Menon, EniferBio
Anaerobic bioreactorHeterotrophs
Mixotrophs
Low cell mass yield
Low capital costs
Low energy consumption
Requires metabolite production and valorization
Non-sterile operation
Most installed industrial capacity
Glucose or glycerol—White Dog Labs
Yeast separation—Fluid Quip Technologies & ICM
Gas bioreactorMethylotrophs
Chemoautotrophs
Mixotrophs
Variable cell mass yield
High capital costs
High energy consumption
Could require metabolite production and valorization
Sterile & non-sterile operation
Limited installed industrial capacity
Methane—Calysta, Unibio, String Bio & Circe Biotechnologie
CO2, H2, and O2—Kiverdi, Novo Nutrients, Deep Branch Biotechnology, Solar Foods, Avecom & LanzaTech
Glucose & syngas—White Dog Labs
Photosynthetic bioreactorPhotoautotrophs
Mixotrophs
High cell mass yield
Low capital costs
High energy consumption
Sterile operation
No known installed capacity
CO2 and light—Bioprocess Algae & Pond Technologies
Open cultivation systemsPhotoautotrophs
Heterotrophs
Mixotrophs
Variable cell mass yield
Low capital costs
Low energy consumption
Non-sterile operation
Limited installed industrial capacity
Brewing by-products—iCell Sustainable Nutrition
Open photosynthetic system—Cellana

Table 24.

Production of SCP.

Source: [148], extended from Refs. [149, 150].

As Table 24 shows, different approaches are being followed. For a review on SCP, see Refs. [85, 151].

A major difficulty with methane is its low solubility in water, resulting in low productivity. Paraffin oil as a “methane vector” for improved mass transfer and higher cell densities was suggested in the literature [64]. Koutinas et al. have proposed to use of γ-alumina pellets to improve methane fermentation [152]. Emulsion-based fermentation to enhance the mass transfer of methane is presented in Ref. [153].

For the bioprocess, Table 25 contrasts two reactor configurations, the “classic” CSTR (continuously stirred tank reactor) and the loop reactor.

Conventional stirred vesselsModern loop reactors
High energy costsLow energy costs
10 kW/m3 (approx.)2 kW/m3 (approx.)
Less defined fluid flow patternQuantified fluid flow pattern
Intrafermentor cooling (jacket)Extrafermentor cooling (plate exchanger)
Complicated heat removalEasy removal of the fermentation heat load
Limited scale-upLess limited scale-up
Indefinite mixing—irregular residence timesMore definite mixing—regular residence times
Cell yield on oxygen of 10–20%, hence requiring a high aeration rateCell yield on oxygen of 40–50%, hence requiring less aeration
Batch and continuous operationContinuous operation
Low yield and productivityHigh yield and productivity
Limited controlEfficient control
Moving parts (easily contaminated)No moving parts
Expensive capital and running costsCheaper to install and run

Table 25.

Fermenter comparison for SCP production.

Source: [51].

For methane fermentation in a sequencing batch bioreactor, see Ref. [154], for batch fermentation, see Ref. [155], and for a cascade of fermenters, see Ref. [156].

The historic Norferm process (see above) used a loop reactor, which can be seen as a PFR (plug flow reactor) comparable to an airlift fermenter only with a different agitation mode of the fluid, using a pump instead of the injected gasses to circulate the medium. Table 26 assesses the relative production costs of different SCP processes.

Capital and manufacturing cost [%] for several SCP processes
Relative costs (units) for:
MethanolEthanolMethanen-Paraffins
Total capital cost10097.6150.8107.7
Total manufacturing cost100229.675.2129.7
Comparative production costs of SCP
ProcessRelative production costs [%] for:
Raw materials
SubstrateOther chemicalsUtilitiesLaborMiscellaneousTotal
Yeast—paraffin29.433.923.88.414.5100
Bacterium—methanol47.426.414.26.25.8100
Yeast—ethanol63.99.912.05.19.1100
Fungus—sulfite liquor17.033.724.811.013.5100
Bacterium—bagasse25.713.536.68.315.9100
Algae—CO2?16.513.815.354.4-
Fungus—lignocellulosic wastes?14.511.322.951.3-

Table 26.

Comparative costs of various SCP processes.

Source: [51].

It is estimated that methane fermentation comes with the highest investment costs (CAPEX), but will allow the lowest operational costs (OPEX), making that feedstock attractive for large-scale operations. When we look at the relative cots for substrate and utilities, there are also marked differences in the various processes. A waste stream has the advantage of low costs, but stable quality and quantity have to be ensured. Utility costs will depend on the geographic location of the site, apart from the unit operations chosen. Cooling costs are determined by the process temperature, and labor costs can be controlled by the degree of plant automation and the complexity of the process. Costs in SCP production are further discussed in Ref. [157].

For engineering factors in the production of SCP, see Refs. [158, 159]. For process development, see Refs. [160].

Downstream processing depends on the exact target product(s). The “classic” separation of cells from the fermentation medium is centrifugation, followed by spray drying. Imasaka et al. have suggested cross-flow filtration of the fermentation broth with ceramic membranes [161], and Yang et al. the continuous methane fermentation in a fixed-bed reactor packed with loofah [69]. Heat treatment or enzymatic treatment [132, 162] can be used for nucleic acid reduction and digestability improvements.

2.2 PHB production process

In Ref. [163], the authors carried out a techno-economic assessment of methanotrophic PHB manufacturing, compare Figure 4.

Figure 4.

PFD (process flow diagram) to convert methane into PHB, with mass balance. Source: [163].

The key findings from Ref. [163] for a PHB production on the order of 100,000 t/a by methanotrophic fermentation were:

  • Downstream process: acetone–water solvent extraction

  • Production costs: $4.1–$6.8/kg PHA

  • Raw material costs reduction compared to sugar feedstock: 22% instead of 30–50%

  • Cooling costs: 28% of the operational costs

  • Strong advantages of using thermophilic methanotrophs—production costs go down to $3.2–5.4/kg PHA [163], because of cheaper cooling (cooling water instead of a refrigerant)

These figures of 3.2–6.8$/kg of PHB are very promising, as today PHA suffers from high production costs and hence a limited market.

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3. Discussion

Today, we are in an “oil economy,” as crude oil is the basis for a major share of energy and materials production. With an accelerating shift toward a circular economy and renewable resources, a paradigm change is about to happen. Biorefineries and biobased products are seeing strong interest from various stakeholders, as do renewable energies, such as wind and solar.

However, when a realistic view is applied, one will quickly see that decarbonization is not that simple. Biomass and hydrogen will play an important role, for sure; the major issue with stepping out of oil is the sheer size of the industry. When one wants to replace the feedstock for 400 million tons of polymers per year, and for hundreds of millions of tons of base chemicals, agricultural resources are simply not sufficiently available, at least not without creating serious disruptions in feed and food production. There is not only a distribution problem of feed and food but a more fundamental land scarcity issue. We are simply not able to convert all “unmanaged” land into fields and pastures to cater to the world population not only for food but also for materials. Neither can be the productivity of the existing land be pushed upwards indefinitely; for fast, secure, and reliable scale-up of SCP, biopolymer, and other materials produced from non-agricultural and non-oil sources, methane from natural gas seems to be the one option.

Based on the circular economy concept, deriving nutrients from (bio)waste is a sustainable approach. For an analysis of SCP made from biowaste as a feed additive using, see Refs. [164, 165].

3.1 SCP

In Table 27, the market of alternative protein sources is summarized.

Protein sourceProduction volume [Mton DM/y]Farm gate price [$/kg DM)Average protein content [% DW]Price per unit protein [$/kg protein DM]
AnimalFish66.72.0715–2010–14
Pork108.51.54207.7
Chicken92.71.43314.6
Beef62.72.702510.8
VegetableSoybean320.20.37351.1
Wheat712.70.19121.6

Table 27.

Different animal and vegetable protein sources compared by their production volumes and prices.

DM = dry matter. Reproduced from [166].

As Table 27 exemplifies, the “farm gate” price of vegetable protein is rather low.

Production volumes of alternative protein from microbes are tabulated (Table 28).

OrganismsProduction volume [ton DM/y]Production costs [€/kg DM]Global market value [Billion €]Yearly growth [% per year]Remarks
Yeast3,000,0009.27.9Mostly commercialized as baker’s yeast and for ethanol fermentation. Global market value projected to 2019
Algae (microalgae)90004–252.410Besides feed and food, derivatives are also used
Mycoprotein (Quorn™)25,0000.214Investments for a plant of 22,000 tons per year were done in 2015
Bacteria (Profloc™)50001–1.1No commercial production yet
Bacteria (FeedKind™)80,000No commercial production yet
Valpromic5000No commercial production yet

Table 28.

Current status of different microbial proteins based on their market size and production volumes.

Source: [166].

Today, yeast is undoubtedly the largest volume SCP source, also for food applications.

In Ref. [167], SCP production by the yeast Kluyveromyces marxianus var marxianus is described.

The FeedKind™ plant is currently under construction [168], Profloc™ went out of business.

When we assume a protein demand per person of 70 g per day of SCP, with a world population of 7.9 billion people, the theoretical market potential for bacterial SCP would be 0.5 million tons per day or 200 million tons per year. At a conversion ratio of 1 g CH4 to 1 g of SCP, we arrive at ∼288 million m3 of methane, which is approximately 7% of today’s natural gas consumption. So if we were to provide all protein for humanity by bacterial SCP, only a fraction of the natural gas stream would be required.

Bacterial single-cell protein has also been envisioned as a possible protein source in a global food catastrophe, where agricultural protein production is suddenly impaired, as elaborated by Juan B. Garcia Martinez [169, 170].

3.2 Bioplastics

Today, bioplastics have a market share of 1–2% of conventional plastics materials. It is estimated that bioplastics could replace 90% of petrochemical plastics, particularly in standard application like packaging (only for high-performance materials, such as PEEK or PFTE, no suitable bioplastics counterparts is yet known to exist). Table 29 takes a look at which bioplastics could replace the most common petrochemical plastics. For instance, LDPE could be replaced to some extent by a “drop in” material of similar property set (bio-PE), and by biopolymers with different characteristics, such as PBAT, PBS, and PHA, to the other part.

Non-biodegradableBiodegradable
Petrochemical PlasticsBioplastics (drop-in, partly biobased)Petrochemical PlasticsBioplastics
Bio-PTTPBAT (can be partly biobased)PBSPHAPLATPSCellulose-based
LDPEbio-PE5510151055
PPbio-PP1051020201520
HDPEbio-PE50101510105
PETbio-PET60105205
PS20302525
PVCbio-PVC502030
EPS7030
PAbio-PA80
PURbio-PUR80201010
Other thermo-plastics101020202020
Other plastics101020202020

Table 29.

Overview of the technical substitution potential of regular plastics (left columns) by bioplastics.

Numbers are in %. Source: [86].

As Table 29 shows, a handful of “drop-in” and degradable bioplastics can replace the most common petrochemical plastics. Overall, it is estimated that up to 90% of conventional polymers can be replaced by biopolymers. The benefits of such a replacement are depicted in Figure 5.

Figure 5.

GWP (global warming potential), land use, and water use of petrochemical and bioplastics packaging materials. Source: [86].

For simplification, the land use of petroplastics is set to zero, as it is negligible. Also, as Figure 7 shows, the water use of petroplastics is low. The error bars indicate the possible range of the figures. According to IfBB [171], the footprint of bioplastics is considerable. For instance, 1 ton of PHB requires 2.86 tons of sugar (glucose) or 3.24 tons of starch for its production. One ton of PLA requires 1.47 tons of sugar or 1.67 tons of starch. Yields of crops differ, for example, 10.03 tons of sugar/ha for sugar beet and 0.83 tons of starch/ha for wheat, resulting in specific land requirements for the materials’ feedstocks, see Figure 6.

Figure 6.

The footprint of the two bioplastics PHB and PLA. Source: [171].

PLA today is the most important bioplastics material, and there is a shortage of supply in the market, leading to a surge in prices. The strong growth is expected to continue. Major producers of PLA are Total/Corbion (Purac™) and Cargill/Natureworks (Ingeo™). The PHB market cannot be considered mature, as the volume is still minute, but there are several established players, see Table 30.

Current industrial production of polyhydroxyalkanoates (PHA)
Company nameCarbon substrateProduct nameProduction [t/a]
Danimer Scientific (formerly Meredian Holdings Group Inc./MHG)Canola oilSeluma™15,000
Metabolix/AntibioticosWitchgrass, camelina, sugar caneMirel, Mvera™10,000
TianAn Biologic Material CoCorn/cassava starchENMAT™10,000
Tianjin GreenBioCorn starchSoGreen™10,000
Bio-onBeet or sugar caneBio-on™10,000
Shenzhen Ecomann Biotech. CoCorn starch5000
PHB IndustrialSugar caneBiocycle™2000
KanekaVegetable oilAonilex™1000
BiomerSugar (sucrose)Biomer P™
Newlight TechnologiesWaste methaneAirCarbon™>500

Table 30.

Players in the PHB market today (status 2016).

Source: [163].

Taking an existing biopolymer and devising a more cost-effective production technology is more likely to bring success than trying to synthesize and/or isolate a totally novel bioplastics material. Hence PLA and PHB are promising materials for methanotrophic fermentation.

Taking 90% of 400 million tons of polymers and a conversion ratio of 1 g of polymer per g of methane, we see that roughly 13% of the global natural gas production would be required to provide the feedstock for the plastics. When we further assume that in the future, a significant share of polymer materials will be recycled, the demand for virgin polymers will be lower, also reducing the fraction of natural gas needed to cater to it.

Scale-up of PHB production by methanotrophic fermentation is discussed in Refs. [172, 173, 174, 175].

3.3 Value-added materials

Likewise for the above-mentioned and further chemicals that can be obtained from methane by fermentation, there is a sound market. Like SCP and biopolymers, they can be produced virtually without requiring (agricultural) land. Decoupling manufacturing from both crude oil and farming activities is attractive as it relieves pressure on existing feed and food value chains. When we estimate that between 500 and 1000 million tons of chemicals are to be made from other sources than oil, at again a rough value of 1 g methane per g of product, we end up with 16–32% of natural gas consumption per year.

3.4 Scenario of methane as sole feedstock

So if we were to produce “all” global demand for protein, plastics, and chemicals from natural gas, which for sure is an exaggerated assumption, we would require approximately 1/3 to 1/2 of the annual natural gas production of today. Let us revisit briefly Figure 7 and add the above estimations to it:

Figure 7.

Graphical depiction of annual natural gas demand for theoretically meeting the entire feedstock demand for SCP, plastics, and chemicals, compared to the natural gas production figures in IEA’s scenarios STEPS, APS, and NZE, which span 3300–4500 billion m3/year annual demand between 2010 and 2030 (compare Figure 7).

Also, biogas could cater to that raw material need. Production volumes are not yet close to 2000 billion m3 per year, but several studies suggest that that potential is within reach.

The authors are firmly convinced that methanotrophic gas fermentation is the key enabling technology to succeed in abandoning crude oil as universal feedstock, by allowing cost-effective and scaleable production of protein, plastics, and chemicals, in a sustainable way, by avoiding the need for immense agricultural and associated production factors, such as water, fertilizer, and pesticides. As Liew et al. have started in their review article [49]—gas fermentation offers a flexible platform for large, industrial-scale production of low-carbon-fuels and chemicals from various feedstocks. This view is shared by other researchers, see Ref. [176].

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4. Summary and outlook

The world faces serious challenges from climate change, growing population, and increasing industrialization. The demand for food skyrockets, as does the desire for energy and various products. The capacity of the world’s oceans to supply fish and of the world’s fields to provide feed and food is limited, and measures to boost productivity have partly been exhausted. Prominent footprint calculations show clearly that the rate with which resources are consumed surpasses the regeneration by a factor of more than 2. Earth exhaustion day is advancing from year to year.

It is well-understood that the current meat production is not sustainable. Cultured meat obviously still needs significant development time to become cost-competitive [177]. Aquaculture can provide fish to the world’s plate despite overfishing, but it needs fodder, which today is often taken from oceans—fish meal and fish oil, or soy, which has its own sustainability issues. Plant-based protein for food is not necessarily sustainable either, considering the large land areas that are required, besides fertilizers, pesticides, etc.

There is an urgent need for large-scale and cheap protein sources that are independent of land use. This has re-sparked interest in microbial protein production, which does not need but very little land and water—feeding microbes sugar or starch virtually perverts any attempts for sustainability: Methane can be a very valid option here. Øverland et al. argued: “In recent years, the increasing global demand for sustainable protein sources, independent of marine origin, agricultural land use, and climatic changes, has led to renewed attention on the potential of microbial protein for use in animal production. Focus has been on methane, the main component of natural gas, which is found widely in nature [28] as an attractive substrate for bacterial protein production. The abundant supply, cheap transportation, and reasonable cost of natural gas indicate that protein production from natural gas could be realistic on a large scale. Using methane-oxidising bacteria as an amino acid source in animal nutrition may spare over-exploited sources of protein suitable for direct human consumption” [65].

Protein from natural gas might, in the unfortunate event of a global food catastrophe, be a vital protein source for several years. As Allfed states: “Human civilization’s food production system is unprepared for global catastrophic risks (GCRs). Catastrophes capable of abruptly transforming global climate, such as supervolcanic eruption, asteroid/comet impact, or nuclear winter, which could completely collapse the agricultural system. Responding by producing resilient foods requiring little to no sunlight is more cost-effective than increasing food stockpiles, given the long duration of these scenarios (6–10 years)” [169]. The preliminary techno-economic analysis revealed that bacterial SCP can be ramped up fast for global food production when needed [169]. Also, bacterial SCP might play a pivotal role in the energy transition, by providing suitable storage capacity for excess renewable energy in a Power2protein setup [178].

We see strong market dynamics in natural gas fermentation these days. This can be inferred from trends in scientific publications, patent applications, and commercial activities, such as pilot or manufacturing plant establishments by several market incumbents. It can be expected that the interest in this set of technologies will be even more in the near and medium-term future, as the key enabling technology for scaling up bioplastics production, protein manufacturing, and chemicals synthesis in a sustainable way, decoupled from agricultural operations.

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Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1. AlSayed A, Fergala A, Eldyasti A. Sustainable biogas mitigation and value-added resources recovery using methanotrophs integrated into wastewater treatment plants. Reviews in Environmental Science and Biotechnology. 2018;17:351-393. DOI: 10.1007/s11157-018-9464-3
  2. 2. Patel SKS, Kondaveeti S, Lee J-K. Repeated batch methanol production from a simulated biogas mixture using immobilized Methylocystis bryophila. Energy. 2017;145:477-485
  3. 3. Spring F. Naturally occurring polyesters. Nature. 1945;155:272. DOI: 10.1038/155272b0
  4. 4. Nawrath C, Poirier Y. Pathways for the synthesis of polyesters in plants: Cutin, suberin, and polyhydroxyalkanoates, Chapter 8. In: Advances in Plant Biochemistry and Molecular Biology. Vol. 1. 2008. pp. 201-239
  5. 5. Harwood JH, Pirt SJ. Quantitative aspects of growth of the methane oxidizing bacterium Methylococcus capsulatus on methane in shake flask and continuous chemostat culture. Journal of Applied Bacteriology. 1972;35:697-607
  6. 6. Rostkowski KH, Criddle CS, Lepech MD. Cradle-to-gate life cycle assessment for a cradle-to-cradle cycle: Biogas-to-bioplastic (and back). Environmental Science and Technology. 2012;46:9822-9829. DOI: 10.1021/es204541w
  7. 7. Rostkowski KH, Pfluger AR, Criddle CS. Stoichiometry and kinetics of the PHB-producing Type II methanotrophs, Methylosinus trichosporium OB3b and Methylocystis parvus OBBP. Bioresource Technology. 2013;132:71-77
  8. 8. Wang J, Salem DR, Sani RK. Microbial polymers produced from methane: Overview of recent progress and new perspectives. Microbial and Natural Macromolecules. 2021. DOI: 10.1016/B978-0-12-820084-1.00006-5
  9. 9. Poirier Y. Polyhydroxyalknoate synthesis in plants as a tool for biotechnology and basic studies of lipid metabolism. Progress in Lipid Research. 2002;41(2):131-155
  10. 10. Park SJ, Kim TW, Lim S-C. Advanced bacterial polyhydroxyalkanoates: Towards a versatile and sustainable platform for unnatural tailor-made polyesters. Biotechnology Advances. 2012;30(6):1196-1206
  11. 11. Babu RP, O’Connor K, Seeram R. Current progress on bio-based polymers and their future trends. Progress in Biomaterials. 2013;2(8):1-16. DOI: 10.1186/2194-0517-2-8
  12. 12. go!PHA. Available from: http://www.gopha.org [Accessed: 13 January 2022]
  13. 13. Myung J, Flanagan JCA, Waymouth RM, Criddle CS. Expanding the range of polyhydroxyalkanoates synthesized by methanotrophic bacteria through the utilization of omega-hydroxyalkanoate co-substrates. AMB Express. 2017;7:118. DOI: 10.1186/s13568-017-0417-y
  14. 14. Yadav B, Pandey A, Rajeshwar BT, Tyagi D, Drogui P. 17—Polyhydroxyalkanoate production from feedstocks: Technological advancements and techno-economic analysis in reference to circular bioeconomy. In: Biomass, Biofuels, Biochemicals, Circular Bioeconomy—Current Status and Future Outlook. Amsterdam, Netherlands: Elsevier. 2021. pp. 477-513
  15. 15. Wingender J, Neu TR, Wingender J, Neu TR, Flemming H-C. Microbial Extracellular Polymeric Substances: Characterization, Structure and Function. Berlin Heidelberg: Springer-Verlag; 1999
  16. 16. Sandford PA, Laskin AI. Extracellular Microbial Polysaccharides. Washington, USA: American Chemical Society. 1978. ISBN: 978-0841203723
  17. 17. Gęsicka A, Oleskowicz-Popiel P, Łężyk M. Recent trends in methane to bioproduct conversion by methanotrophs. Biotechnology Advances. 2021;53 Article 107861
  18. 18. Production of lactic acid from organic waste or biogas or methane using recombinant methanotrophic bacteria. EP 9.3129513. 15.02.2017
  19. 19. Method for producing isoprene using recombinant halophilic methanotroph. SK Innovation Co.; June 12, 2018. US 9994869
  20. 20. Methanotrophic mutant strain for producing squalene. KR 1020180124189. 21.11.2018
  21. 21. Production of succinic acid from organic waste or biogas or methane using recombinant methanotrophic bacterium. WO/2015/155791. 15.10.2015
  22. 22. Methanobactin: A copper binding compound having antibiotic and antioxidant activity isolated from methanotrophic bacteria. US 75.20040171519. 02.09.2004
  23. 23. Cantera S, Lebrero R, Muñoz R. Ectoine bio-milking in methanotrophs: A step further towards methane-based bio-refineries into high added-value products. Chemical Engineering Journal. 2017;328:44-48
  24. 24. Cantera S, Lebrero R, Rodríguez E, García-Encina PA, Munoz R. Continuous abatement of methane coupled with ectoine production by Methylomicrobium alcaliphilum 20Z in stirred tank reactors: A step further towards greenhouse gas biorefineries. Journal of Cleaner Production. 2017;152:134-141
  25. 25. Henard CA, Akberdin IR, Kalyuzhnaya MG, Guarnieri MT. Muconic acid production from methane using rationally-engineered methanotrophic biocatalysts. Green Chemistry. 2019;21:6731-6737. DOI: 10.1039/C9GC03722E
  26. 26. Ye RW, Yao H, Stead K, Wang T, Tao L, Cheng Q, et al. Construction of the astaxanthin biosynthetic pathway in a methanotrophic bacterium Methylomonas sp. strain 16a. Journal of Industrial Microbiology and Biotechnology. 2007;34(4):289. DOI: 10.1007/s10295-006-0197-x
  27. 27. Strong PJ, Kalyuzhnaya M, Clarke WP. A methanotroph-based biorefinery: Potential scenarios for generating multiple products from a single fermentation. Bioresource Technology. 2016;215:314-323
  28. 28. Lee SY, Cho JM, Chang YK, Oh Y-K. Cell disruption and lipid extraction for microalgal biorefineries: A review. Bioresource Technology. 2017;244:1317-1328
  29. 29. Nitsos C, Filali R, Taidi B, Lemaire J. Current and novel approaches to downstream processing of microalgae: A review. Biotechnology Advances. 2020;45:107650
  30. 30. DaSilva EJ, Ratledge C, Sasson A. Biotechnology: Economic and social aspects (issues for developing countries). The Economic Viability of Single Cell Protein (SCP) Production in the Twenty-First Century; Cambridge, UK: Cambridge University Press. 1992
  31. 31. Babel W. Ullmann’s Encyclopedia of Industrial Chemistry. Single Cell Proteins; Weinheim, Germany: Wiley VCH. 2000
  32. 32. SRL S, Beveridge EG, Elton C, Danielli JF, Booth IR, Herbert HB. New horizons in industrial microbiology. Single cell protein [and discussion]. Philosophical Transactions of the Royal Society of London Series B Biological Sciences (1934–1990). 1980
  33. 33. Ali S, Mushtaq J, Nazir F, Sarfraz H. Production and processing of single cell protein (SCP)—A review. European Journal of Pharmaceutical and Medical Research. 2017;4:86-94
  34. 34. Bewersdorff M, Dostálek M. The use of methane for production of bacterial protein. Biotechnology and Bioengineering. 1971. DOI: 10.1002/bit.260130104
  35. 35. Calcott PH. Continuous Cultures Of Cells. Boca Raton, Florida, USA: CRC Press; 1981 ISBN: 9781000694574
  36. 36. Koo CW, Rosenzweig AC. Biochemistry of aerobic biological methane oxidation. Chemical Society Reviews. 2021;50:3424-3436. DOI: 10.1039/D0CS01291B
  37. 37. Schøyen HF, Svihus B, Storebakken T, Skrede A. Bacterial protein meal produced on natural gas replacing soybean meal or fish meal in broiler chicken diets. Archives of Animal Nutrition. 2007;61(4):276-291. DOI: 10.1080/17450390701431953
  38. 38. Production of single cell protein by methanol-using bacteria. Journal of Fermentation Technology. 1986;64(2):97-186
  39. 39. Single-cell protein. Safety for Animal and Human Feeding. Proceedings of the Protein-Calorie Advisory Group of the United Nations System Symposium “Investigations on Single-Cell Protein” held at the Istituto di Ricerche Farmacologiche ‘Mario Negri’ Milan, Italy, March 31–April 1, 1977. ISBN: 0-08-023764-9
  40. 40. Liquid protein diets. Hearing Before The Subcommittee on Health and The Environment of The Committee on Interstate and Foreign Commerce Ninety-Fifth Congress. First Session To Provide Consumers With Better Information On The Most Popular Diet In America Today. Liquid. Serial No. 95-83. December 28, 1977
  41. 41. Opinion of the Scientific Panel on additives and products or substances used in animal feed (FEEDAP) on the safety of BioProtein: Product of fermentation from natural gas. 4 July 2005. Available from: https://www.efsa.europa.eu/en/efsajournal/pub/230. doi:10.2903/j.efsa.2005.230
  42. 42. Giec A, Skupin J. Single cell protein as food and feed. Molecular Nutrition & Food Research. 1988;32(3):219–229
  43. 43. Sherwood M. Single-cell protein comes of age. New Scientist. 1974;28(11):634-639
  44. 44. Bratosin BC, Darjan S, Vodnar DC. Single cell protein: A potential substitute in human and animal nutrition. Sustainability. 2021;13:9284. DOI: 10.3390/su13169284
  45. 45. Ravindra AP. Value-added food: Single cell protein. Biotechnology Advances. 2000;18:459-479
  46. 46. Patias LD, Maroneze MM, Siqueira SF, de Menezes CR, Zepka LQ, Jacob-Lopes E. Single-cell protein as a source of biologically active ingredients for the formulation of antiobesity foods. Alternative and Replacement Foods. DOI: 10.1016/B978-0-12-811446-9.00011-3
  47. 47. Lipids from methanotrophic bacteria for cholesterol reduction. 05.04.2006. EP 79.1641475
  48. 48. Vahidi H, Mojab F, Taghavi N. Effects of carbon sources on growth and production of antifungal agents by Gymnopilus spectabilis. Iranian Journal of Pharmaceutical Research. 2006;3:219-222
  49. 49. Liew FM, Martin ME, Tappel RC, Heijstra BD, Mihalcea C, Köpke M. Gas fermentation—A flexible platform for commercial scale production of low-carbon-fuels and chemicals from waste and renewable feedstocks. Frontiers in Microbiology. 2016;7:694. doi: 10.3389/fmicb.2016.00694
  50. 50. Kunasundari B, Murugaiyah V, Kaur G, Maurer FHJ, Sudesh K. Revisiting the single cell protein application of Cupriavidus necator H16 and recovering bioplastic granules simultaneously. PLoS ONE. 2013;8(10):e78528. Available from: www.plosone.org
  51. 51. Goldberg I. Single Cell Protein. New York, USA: Springer; 1985. ISBN: 978-3-642-46542-0. DOI: 10.1007/978-3-642-46540-6
  52. 52. Available from: https://www.heise.de/tp/features/Zur-Geschichte-einer-ehemaligen-Zukunftstechnologie-die-noch-nicht-abgehakt-ist-4221160.html?seite=all
  53. 53. Egorov I, Kupina L, Aksyuk I, Murtazaeva R. Gaprin—A protein source. Ptitsevodstvo. 1990;8:25-27
  54. 54. Chirikov S, Shkirin A, Savchenko I, Bunkin N, Diuldin M. Assessment of the possibility of identifying aqueous suspensions of protein-containing particles by the light scattering matrix. In: ECOBALTICA 2019, IOP Conf. Series: Earth and Environmental Science. 2019;390:012030. doi:10.1088/1755-1315/390/1/012030
  55. 55. Chirikov SN, Shkirin AV, Konnova AS. Study of light-scattering properties of protein-containing microparticles with a small difference in refractive indices. Journal of Physics. 2020
  56. 56. Kent JA. Riegel’s Handbook of Industrial Chemistry. Springer; 1992 ISBN: 9781475764338
  57. 57. Champagnat A, Vernet C, Lainé B, Filosa J. Biosynthesis of protein–vitamin concentrates from petroleum, Nature, 1963;197:13–14
  58. 58. Champagnat A. Protein from petroleum. Scientific American. 1965;213(4):13-17
  59. 59. Alani DI, Moo-Young M. Single cell protein production from petroleum derivatives and its utilization as food and feed. Perspectives in Biotechnology and Applied Microbiology. 1986;1-6. DOI: 10.1007/978-94-009-4321-6_1
  60. 60. Ismail WA, Van Hamme, Jonathan D, Kilbane JJ, Gu J-D. Editorial: Petroleum Microbial Biotechnology: Challenges and Prospects
  61. 61. Henk G, Heinz T. Hydrocarbon content in the fat of meat pigs after feeding “Fermosin”. Arch Tierernahr. 1993;45(1):35-47. DOI: 10.1080/17450399309386086
  62. 62. Alani DI, Moo-Young M. The economical aspects of single cell protein production from petroleum derivatives. Perspectives in Biotechnology and Applied Microbiology. 1986
  63. 63. Regulations Commission Regulation (EU) No. 68/2013 of 16 January 2013 on the Catalogue of feed materials (text with EEA relevance)
  64. 64. Han B, Tao S, Hao W, Gou Z, Xing X-H, Jiang H, et al. Paraffin oil as a “methane vector” for rapid and high cell density cultivation of Methylosinus trichosporium OB3b. Applied Microbiology and Biotechnology. 2009;83:669-677. DOI: 10.1007/s00253-009-1866-2
  65. 65. Øverland M, Tauson A-H, Shearer K, Skrede A. Evaluation of methane-utilising bacteria products as feed ingredients for monogastric animals. Archives of Animal Nutrition. 2010;64(3):171-189. DOI: 10.1080/17450391003691534
  66. 66. Nevalainen H, editor. Grand Challenges in Fungal Biotechnology. New York, USA: Springer ISBN: 978-3-030-29540-0; 2020. DOI: 10.1007/978-3-030-29541-7
  67. 67. Knight N, Roberts G, Shelton D. The thermal stability of Quorn™ pieces. International Journal of Food Science & Technology. 2001;36(1):47-52
  68. 68. ICI to scale up single cell protein process, Chemical & Engineering News, 1976;54(42):25. DOI: 10.1021/cen-v054n042.p025
  69. 69. Rose AH. The microbiological production of food and drink. Scientific American. 1981;245(3):126-134. DOI: 10.1038/scientificamerican0981-126
  70. 70. Jenkins G. SCP—The BP protein process. In: Greenshields R, editor. Resources and Applications of Biotechnology. 1988
  71. 71. Verrier D, Morfaux JN, Albagnac G, Touzel JP. The French programme on methane fermentation. Biomass. 1982
  72. 72. Damron BL, Simpson CF, Eldred AR, Harms RH. Liquipron as a protein source for broilers. Poultry Science. 1979;58:247-249
  73. 73. Available from: https://www.equinor.com/en/news/archive/1999/02/17/BioproteinBegins.html
  74. 74. Olsen DF, Jørgensen JB, Villadsen J, Jørgensen SB. Modeling and simulation of single cell protein production. In: Banga JR, Bogaerts P, Van Impe J, Dochain D, Smets I, editors. Proceedings of the 11th International Symposium on Computer Applications in Biotechnology (CAB 2010), Vol. 43(6); July 7–9, 2010; Leuven, Belgium; IFAC Proceedings Volumes. 2010. pp. 502-507
  75. 75. Nunes JJ. Theoretical energy requirements of single cell protein production from methanol and methane using metabolic flux analysis. In: Proceedings of the Tobago Gas Technology Conference (TGTC) 2008, Conference Paper. 2008
  76. 76. Opinion on the safety of BioProtein® by the Scientific Panel on Animal Feed of the Norwegian Scientific Committee for Food Safety, Revised version. Adopted on the 5th of October 2006, VKM Report 2006: 43
  77. 77. Perlman D. Annual reports on fermentation processes. Annual Reports on Fermentation Processes. Cambridge, Massachusetts, USA: Academic Press. 1977. pp. 1. ISBN: 0-12-040301-3
  78. 78. James K. Single cell protein process targeted for licensing. Chemical & Engineering News. 1983;61. DOI: 10.1021/cen-v061n031.p021
  79. 79. Single Cell Protein Process Targeted for Licensing. August 1, 1983 C&EN, 21
  80. 80. Israelidis CJ. Nutrition-single cell protein, twenty years later. Available from: https://www.semanticscholar.org/paper/NUTRITION-SINGLE-CELL-PROTEIN-%2C-TWENTY-YEARS-LATER-Israelidis/cd52ecd95eb39b199b3fc33893fda2b0cc77ac07
  81. 81. Katsnelson A. Dow pulls out of bioplastics due to slow sector maturation. Nature Biotechnology. 2005;23:638
  82. 82. Tannenbaum SR, Wang DIC, editors. Single-Cell Protein II. MIT Press; 1975. ISBN: 0-262-20030-9
  83. 83. Hamer G, Harrison DEF. Single cell protein: The technology, economics and future potential. In: Harrison DEF, Higgins IJ, Watkinson R, editors. Hydrocarbons in Biotechnology. London: Heyden & Son, Ltd.; 1980. pp. 59-73
  84. 84. Mateles RI, Tannenbaum SR, editors. Single-Cell Protein. Cambridge, Massachusetts: MIT Press; 1968
  85. 85. Cooney CL, Rha C, Tannenbaum SR. Single-cell protein: Engineering, economics, and utilization in foods. Advances in Food Research. 1980;26:1-52
  86. 86. Brizga J, Hubacek K, Feng K. The unintended side effects of bioplastics: Carbon, land, and water footprints. One Earth. 2020;3(1):45-53
  87. 87. Kuzniar A, Furtak K, Włodarczyk K, Stepniewska Z, Wolinska A. Methanotrophic bacterial biomass as potential mineral feed ingredients for animals. International Journal of Environmental Research and Public Health. 2019;16:2674. DOI: 10.3390/ijerph16152674
  88. 88. Gerth K, Trowitzsch W, Piehl G, Schultze R, Lehmann J. Inexpensive media for mass cultivation of myxobacteria. Applied Microbiology and Biotechnology. 1984;19:23-28
  89. 89. Núñez Decap M, Ballerini Arroyo A, Alarcón Énos J. Evaluation of single cell protein from yeast for the development of wood adhesives. European Journal of Wood and Wood Products. 2016;74:821-828. DOI: 10.1007/s00107-016-1063-9
  90. 90. Yazdian F, Hajizadeh S, Shojaosadati SA, Khalilzadeh R, Jahanshahi M, Nosrati M. Production of single cell protein from natural gas: Parameter optimization and RNA evaluation. Iranian Journal of Biotechnology. 2005;3(4):235
  91. 91. Bhalla TC, Mehta PK, Bhatia SK, Pratush A. Microorganisms for food and feed. In: Fundamentals of Food Biotechnology. New Delhi, India: Anne Publisher; 2009. Available from: https://www.researchgate.net/publication/303941522_Microorganism_for_food_and_Feed
  92. 92. Taran M, Asadi N. A novel approach for environmentally friendly production of single cell protein from petrochemical wastewater using a halophilic microorganism in different conditions. Petroleum Science and Technology. 2014;32:625-630
  93. 93. Zha X, Tsapekos P, Zhu X, Khoshnevisan B, Lu X, Angelidaki I. Bioconversion of wastewater to single cell protein by methanotrophic bacteria. Bioresource Technology. 2021;320:124351
  94. 94. Rasouli Z, Valverde-Pérez B, D’Este M, De Francisci D, Angelidaki I. Nutrient recovery from industrial wastewater as single cell protein by a co-culture of green microalgae and methanotrophs. Biochemical Engineering Journal. 2018;134:129-135
  95. 95. Xu M, Zhou H, Yang X, Angelidaki I, Zhang Y. Sulfide restrains the growth of Methylocapsa acidiphila converting renewable biogas to single cell protein. Water Research. 2020;184:116138
  96. 96. Yazdian F, Hajizadeh S, Shojaosadati SA, Khalilzadeh R, Jahanshahi M, Nosrati M. Production of single cell protein from natural gas: Parameter optimization and RNA evaluation. Iranian Journal of Biotechnology. 2005;3:235-242
  97. 97. Marstrand PK. Production of microbial protein: A study of the development and introduction of a new technology. Research Policy. 1981;10(2):148-171
  98. 98. Frydendahl Hellwing AL. Bacterial protein meal as protein source for monogastric animals—Comparative studies on protein and energy, metabolism [PhD thesis]. Frederiksberg, Denmark: Samfundslitteratur Grafik; 2005. ISBN: 87-7611-110-5
  99. 99. Volesky B, Zajic JE, Carroll KK. Author notes: Feeding studies in rats with high protein fungus grown on natural gas. The Journal of Nutrition. 1975;105(3):311-316. DOI: 10.1093/jn/105.3.311
  100. 100. Walz OP, Brune H. Zeitschrift für Tierphysiologie Tierernährung und Futtermittelkunde. Proteinverwertung und Wirkung von Methioninergänzung bei Einzellerbiomasse aus Methylomonas clara gemessen mit Ratten und Absetzferkeln. January–August 1984. doi:10.1111/j.1439-0396.1984.tb01428.x
  101. 101. Hanssen JT. Bioproteins in the feeding of growing–finishing pigs in Norway: I. Chemical composition, nutrient digestibility and protein quality of “Pruteen”, “Toprina”, “Pekilo” and a methanol-based yeast product (Pichia aganobii). Journal of Animal Physiology and Animal Nutrition. 1981;46(1-5):182-196
  102. 102. Becker PM. Single cell proteins in diets for weanling pigs. Report 03/0016848. Animal Sciences Group-Nutrition & Food
  103. 103. Stiglmair-Herb MT, Pospischil A. Enzymhistochemische Untersuchungen am Darmepithel von Hunden nach Kasein-Diät, Single Cell Protein-Diät und konventioneller Fütterung. Transboundary and Emerging Diseases. 1985;32(1-10):764-771
  104. 104. Teller E, Godeau J-M. Evaluation of the nutritive value of single-cell protein (Pruteen) for lactating dairy cows. The Journal of Agricultural Science. 1986;106(3):593-599. DOI: 10.1017/S0021859600063462
  105. 105. Van Weerden EJ, Huisman J. Digestibility of protein and amino acids of a fermentation single-cell protein for veal calves. Animal Feed Science and Technology. 1977;1(4):377-383
  106. 106. Zamani A, Khajavi M, Nazarpak MH, Gisbert E. Evaluation of a bacterial single-cell protein in compound diets for rainbow trout (Oncorhynchus mykiss) fry as an alternative protein source. Animals. 2020;10:1676. DOI: 10.3390/ani10091676
  107. 107. Jintasataporn O, Chumkam S, Triwutanon S, LeBlanc A, Sawanboonchun J. Effects of a single cell protein (Methylococcus capsulatus, Bath) in Pacific White Shrimp (Penaeus vannamei) diet on growth performance, survival rate and resistance to Vibrio parahaemolyticus, the causative agent of acute hepatopancreatic necrosis disease. Frontiers in Marine Science. 2021;8:764042. DOI: 10.3389/fmars.2021.764042
  108. 108. Chen Y, Chi S, Zhang S, Dong X, Yang Q, Liu H, et al. Replacement of fish meal with Methanotroph (Methylococcus capsulatus, Bath) bacteria meal in the diets of Pacific white shrimp (Litopenaeus vannamei). Aquaculture. 2021;541:736801
  109. 109. Kabwe M, Chama H, Liang H, Ke J. Methanotroph (Methylococcus capsulatus, Bath) as an alternative protein source for genetically improved farmed tilapia (GIFT: Oreochromis niloticus) and its effect on antioxidants and immune response. Aquaculture Reports. 2021;21. Article 100872
  110. 110. Davies SJ, Wareham H. A preliminary evaluation of an industrial single cell protein in practical diets for tilapia (Oreochromis mossambicus Peters). Aquaculture. 1988
  111. 111. Biswas A, Takakuwa F, Tanaka H. Methanotroph (Methylococcus capsulatus, Bath) bacteria meal as an alternative protein source for Japanese yellowtail, Seriola quinqueradiata. Aquaculture. 2020;529. Article 735700
  112. 112. Sisman T, Gur O, Dogan N, Ozdal M, Algur OF, Ergon T. Single-cell protein as an alternative food for zebrafish, Danio rerio: A toxicological assessment. Toxicology and Industrial Health. 2013
  113. 113. Glencross BD, Huyben D, Schrama JW. The application of single-cell ingredients in aquaculture feeds—A review. Fishes. 2020;5:22. DOI: 10.3390/fishes5030022
  114. 114. Tlusty M, Rhyne A, Szczebak JT, Bourque B, Bowen JL, Burr G, et al. A transdisciplinary approach to the initial validation of a single cell protein as an alternative protein source for use in aquafeeds. PeerJ. 2017;5:e3170. DOI: 10.7717/peerj.3170
  115. 115. Christensen HR, Larsen LC, Frøkiær H. The oral immunogenicity of BioProtein, a bacterial single-cell protein, is affected by its particulate nature. British Journal of Nutrition. 2003;90(1):169-178. DOI: 10.1079/BJN2003863
  116. 116. Litchfield JH. Production of single-cell protein for use in food or feed. In: Microbial Technology. 2nd ed. Academic Press, Inc.; 1979. ISBN: 0-12-551501-4
  117. 117. Available from: http://www.freienohler.de/index.php/freienohl/geschichte/16-geschichte/481-die-wildshauser-holz-bzw-leberwurst-1943-1949.html
  118. 118. Ritala A, Häkkinen ST, Toivari M, Wiebe MG. Single cell protein—State-of-the-art, industrial landscape and patents 2001–2016. Frontiers in Microbiology. 2017
  119. 119. Romarheim OH, Landsverk T, Mydland LT, Skrede A, Øverland M. Cell wall fractions from Methylococcus capsulatus prevent soybean meal-induced enteritis in Atlantic salmon (Salmo salar). Aquaculture. 2013;402–403:13-18
  120. 120. Skrede A, Faaland Schøyen H, Svihus B, Storebakken T. The effect of bacterial protein grown on natural gas on growth performance and sensory quality of broiler chickens. Canadian Journal of Animal Science. 2013
  121. 121. EU is one step closer to the use of PAPs in animal feed—All about feed. Available from: https://www.allaboutfeed.net/animal-feed/raw-materials/eu-is-one-step-closer-to-the-use-of-paps-in-animal-feed/
  122. 122. Bovine spongiform encephalopathy (BSE). EFSA (europa.eu). Available from: https://www.efsa.europa.eu/en/topics/topic/bovine-spongiform-encephalopathy-bse
  123. 123. Skrede A, Mydland LT, Øverland M. Effects of growth substrate and partial removal of nucleic acids in the production of bacterial protein meal on amino acid profile and digestibility in mink. Journal of Animal and Feed Sciences. 2009;18:689-698
  124. 124. Li S, Jiang W, Li M. Oral delivery of bacteria: Basic principles and biomedical applications. Journal of Controlled Release. 2020;327:801-833
  125. 125. Liong M-T, editor. Beneficial Microorganisms in Food and Nutraceuticals. Basel, Switzerland: Springer International Publishing; 2015
  126. 126. Microorganisms in Foods 6: Microbial Ecology of Food Commodities (Microorganisms in Foods). 2nd ed. International Commission on Microbiological Specifications of Foods (ICMSF); 2005 ISBN: 9780387288017
  127. 127. Microorganisms in Foods 7: Microbiological Testing in Food Safety Management. 2nd ed, ISBN: 9783319684604. International Commission on Microbiological Specifications for Foods (ICMSF); 2018
  128. 128. International Commission on Microbiological Specifications for Foods. Microorganisms in Foods 8: Use of Data for Assessing Process Control and Product Acceptance. USA, ISBN: 9781441993748: Springer; 2011
  129. 129. Koivurinta J, Kurkela R, Koivistoinen P. Uses of Pekilo, a microfungus biomass from Paecilomyces varioti in sausage and meat balls. Journal of Food Science and Technology. 1979;14:561-570
  130. 130. Available from: https://www.eniferbio.fi/eniferbio-received-a-e1-2m-blueinvest-grant-by-the-european-maritime-and-fisheries-fund-emff/
  131. 131. LaTurner ZW, Bennett GN, San K-Y, Stadler Lauren B. Single cell protein production from food waste using purple non-sulfur bacteria shows economically viable protein products have higher environmental impacts. Journal of Cleaner Production. 2020
  132. 132. Vogel A, May O. Industrial Enzyme Applications. Wiley–VCH, ISBN: 9783527813780; 2019
  133. 133. Abrahamsson L, Hambraeus L, Hofvander Y, Vahlquist B. Single cell protein in clinical testing, a tolerance test in healthy adult subjects comprising biochemical, clinical and dietary evaluation. Nutrition Metabolism. 1971;13:186-199
  134. 134. Steinmann J, Wottge H-U, Müller-Ruchholtz W. Immunogenicity testing of food proteins: In vitro and in vivo trials in rats. International Archives of Allergy & Applied Immunology. 1990;91:62-65
  135. 135. Hedenskog G, Ebbinghaus L. Reduction of the nucleic acid content of single-cell protein concentrates. Biotechnology and Bioengineering. 1972;XIV:447-457
  136. 136. Jonas DA, Elmadfa I, Engel K-H, Heller KJ, Kozianowski G, König A, et al. Safety considerations of DNA in food. Annals of Nutrition & Metabolism. 2001;45:235-254
  137. 137. Abou-Zeid A-ZA, Khan JA, Abulnaja KO. On methods for reduction of nucleic acids content in a single-cell protein from gas oil. Bioresource Technology. 1995;52:21-24
  138. 138. Available from: https://solarfoods.fi/our-news/solein-submitted-to-the-european-commission-for-novel-food-approval/
  139. 139. Ekenvall L, Dölling B, Göthe C-J, Ebbinghaus L, Von Stedingk L-V, Wasserman J. Single cell protein as an occupational hazard. British Journal of Industrial Medicine. 1983
  140. 140. Kundiyana DK, Huhnke RL, Maddipati P, Atiyeh HK, Wilkins MR. Feasibility of incorporating cotton seed extract in Clostridium strain P11 fermentation medium during synthesis gas fermentation. Bioresource Technology. 2010;101(24):9673-9680
  141. 141. Takashima M, Speece RE. Mineral nutrient requirements for high-rate methane fermentation of acetate at low SRT. Research Journal of the Water Pollution Control Federation. 1989;61(11/12):1645-1650
  142. 142. Takashima M, Speece RE, Parkin GF. Mineral requirements for methane fermentation. Critical Reviews in Environmental Control. 1990:465-479. DOI: 10.1080/10643389009388378
  143. 143. Paek K-Y, Murthy HN, Zhong J-J, editors. Production of Biomass and Bioactive Compounds using Bioreactor Technology. Netherlands: Springer; 2014
  144. 144. Ye Q, Bao J, Zhong J-J. Bioreactor Engineering Research and Industrial Applications. I: Cell Factories. Berlin Heidelberg: Springer-Verlag; 2016 ISBN: 9783662491614
  145. 145. Kadic E, Heindel TJ. An Introduction to Bioreactor Hydrodynamics and Gas-Liquid Mass Transfer. Weinheim, Germany: Wiley, ISBN: 9781118104019; 2014
  146. 146. Liao Q, Chang J-s, Herrmann C, Xia A. Bioreactors for Microbial Biomass and Energy Conversion. Singapore, ISBN: 9789811076770: Springer; 2018
  147. 147. Reisman HB. Economic Analysis of Fermentation Processes. Boca Raton, Florida, USA: CRC Press LLC, ISBN: 9780429553165; 2019
  148. 148. Jones SW, Karpol A, Friedman S, Maru BT, Tracy BP. Recent advances in single cell protein use as a feed ingredient in aquaculture. Current Opinion in Biotechnology. 2020;61:189-197
  149. 149. Available from: Strategies for success in single-cell protein production (luxresearchinc.com), https://www.luxresearchinc.com/blog/strategies-for-success-in-single-cell-protein-production
  150. 150. F3 FIN: Feed Innovation Network. Available from: https://f3fin.org/
  151. 151. Nasseri AT, Rasoul-Amini S, Morowvat MH, Ghasemi Y. Single cell protein: Production and process. American Journal of Food Technology. 2011;6(2):103-116
  152. 152. Koutinas AA, Toutoutzidakis G, Kana K, Kouinis I. Methane fermentation promoted by γ-alumina pellets. Journal of Fermentation and Bioengineering. 1991;72(1):64-67
  153. 153. Myung J, Kim M, Tang SKY. Low energy emulsion-based fermentation enabling accelerated methane mass transfer and growth of poly(3-hydroxybutyrate)-accumulating methanotrophs. Bioresource Technology. 2016;207:302-307
  154. 154. Buczkowska A, Witkowska E, Górski Ł, Zamojska A, Szewczyk KW, Wróblewski W, et al. The monitoring of methane fermentation in sequencing batch bioreactor with flow-through array of miniaturized solid state electrodes. Talanta. 2010;81(4-5):1387-1392
  155. 155. Ahmad MN, Holland CR. Growth kinetics of single-cell protein in batch fermenters. Journal of Food Engineering. 1995;26(4):443-452
  156. 156. Kianoush K-D, Fatemeh Y, Hamid R, Neda MB, Mohsen M, Soheil RM, et al. Simulation of bioreactors for poly(3-hydroxybutyrate) production from natural gas. Iranian Journal of Chemistry and Chemical Engineering. 2020;39(1):313-336
  157. 157. Moo-Young M. Economics of single cell protein production. Process Biochemistry. 1977;12:6
  158. 158. Labuza TP, Le Roux JP, Fan TS, Tannenbaum SR. Engineering factors in single-cell protein production. II. Spray drying and cell viability. Biotechnology and Bioengineering. 1970;12(1):135-140
  159. 159. Labuza TP, Santos DB, Roop RN. Engineering factors in single-cell protein production. I. Fluid properties and concentration of yeast by evaporation. Biotechnology and Bioengineering. 1970;12(1):123-134
  160. 160. Abbott BJ, Clamen A. The relationship of substrate, growth rate, and maintenance coefficient to single cell protein production. Biotechnology and Bioengineering. 1973;15:117. Available from: https://onlinelibrary.wiley.com/doi/abs/10.1002/bit.260150109
  161. 161. Imasaka T, Kanekuni N, So H, Yoshino S. Cross-flow filtration of methane fermentation broth by ceramic membranes. Journal of Fermentation and Bioengineering. 1989;68(3):200-206
  162. 162. Kuddus M. Enzymes in Food Biotechnology: Production, Applications, and Future Prospects. Cambridge, Massachusetts, USA: Academic Press. 2019. ISBN: 9780128132807
  163. 163. Levett I, Birkett G, Davies N, Bell A, Langford A, Laycock B, et al. Techno-economic assessment of poly-3-hydroxybutyrate (PHB) production from methane—The case for thermophilic bioprocessing. Journal of Environmental Chemical Engineering. 2016;4:3724-3733
  164. 164. Tsapekos P, Angelidaki XZI. Proteinaceous methanotrophs for feed additive using biowaste as carbon and nutrients source. Bioresource Technology. 2020;313 Article 123646
  165. 165. Tomlinson EJ. The production of single-cell protein from strong organic waste waters from the food and drink processing industries—2. The practical and economic feasibility of a non-aseptic batch culture. Water Research. 1976;10(5):367-371
  166. 166. Matassa S, Boon N, Pikaar I, Verstraete W. Microbial protein: Future sustainable food supply route with low environmental footprint. Microbial Biotechnology. 2016;9(5):568-575
  167. 167. Anderson PJ, McNeil KE, Watson K. Thermotolerant single cell protein production by Kluyveromyces marxianusvar. marxianus. Journal of Industrial Microbiology & Biotechnology. 1988;3:9-14
  168. 168. Digital signing ceremony unveils location of Calysseo’s world-first commercial FeedKind® plant—FeedKind®. Available from: http://www.feedkind.com/digital-signing-ceremony-unveils-location-calysseos-world-first-commercial-feedkind-plant/
  169. 169. Garcia Martinez JB, Alvarado KA, Denkenberger DC. Synthetic fat from petroleum as a resilient food for global catastrophes: Preliminary techno-economic assessment and technology roadmap. Chemical Engineering Research and Design. 2022;177:255-272. DOI: 10.1016/j.cherd.2021.10.017. PII: S0263-8762(21)00427-5
  170. 170. Throup J, Garcia Martinez JB, Bals B, Cates J, Pearce JM, Denkenberger DC. Rapid repurposing of pulp and paper mills, biorefineries, and breweries for lignocellulosic sugar production in global food catastrophes. Food and Bioproducts Processing. 2022;131:22-39. DOI: 10.1016/j.fbp.2021.10.012. PII: S0960-3085(21)00162-0
  171. 171. Biopolymers—Facts and statistics. Ausgabe 2021. Available from: https://www.ifbb-hannover.de/files/IfBB/downloads/faltblaetter_broschueren/f+s/Biopolymers-Facts-Statistics-einseitig-2021.pdf
  172. 172. Lackner M. Methane as emerging raw material for biopolymers biobased polymers. In: 4th World Congress on Bio-Polymers and Polymer Chemistry, Webinar, Keynote Speech; 30 March. 2021
  173. 173. Lackner M. Methane as sustainable biopolymer feedstock—Bioprocess engineering using methanotrophic bacteria. In: Renewable Resources and Biorefineries Conference, RRB2021; Aveiro, Portugal; September. 2021. Available from: https://www.rrbconference.com/%20RRB
  174. 174. Morais AMMB, Morais RMSC, Drew D, Mustakhimov I, Lackner M. Biodegradable Bio-based Plastics Toward Climate Change Mitigation. New York, USA: Springer. 2021. pp. 1-43. DOI: 10.1007/978-1-4614-6431-0_91-2
  175. 175. Lackner M, Mustakhimov I, Drew D. Feedstock considerations for world-scale PHA production: Methane as viable option. In: 2nd PHA platform World Congress; September 23. 2021 14:15–14:40. Available from: https://www.bioplasticsmagazine.com/en/event-calendar/termine/2nd-pha-world-congress-2020/#anchor_95c7b84a_Accordion-Programme
  176. 176. Pieja AJ, Morse MC, Cal AJ. Methane to bioproducts: The future of the bioeconomy? Current Opinion in Chemical Biology. 2017;41:123-131
  177. 177. Available from: https://www.foodnavigator-usa.com/Article/2021/03/15/When-will-cell-cultured-meat-reach-price-parity-with-conventional-meat
  178. 178. Daneels R. Protein at farm scale from feed compatible components. In: Power to Gas to Protein. Utrecht: Innovation Network; 2016. ISBN: 978-90-5059-524-7. Report no. 15.2.334, May

Written By

Maximilian Lackner, David Drew, Valentina Bychkova and Ildar Mustakhimov

Reviewed: 23 March 2022 Published: 19 May 2022