Open access

The Current Status and Future Expectations in Industrial Production of Lactic Acid by Lactic Acid Bacteria

Written By

Sanna Taskila and Heikki Ojamo

Submitted: November 9th, 2011 Published: January 30th, 2013

DOI: 10.5772/51282

Chapter metrics overview

9,343 Chapter Downloads

View Full Metrics

1. Introduction

Conversion of carbohydrates to lactic acid is one of the most employed fermentation processes in food industry. Applications of lactic acid fermentation are found in dairy industry, production of wine and cider, production of fermented vegetable products and meat industry.

The main markets for lactic acid have been in food, pharmaceutical and cosmetics industry, but presently the main growing application of lactic acid is in the production of biodegradable and renewable raw material based poly lactic acid (PLA) polymers. Production of lactate esters (e.g. butyl lactate) is another growing application as environmentally friendly solvents [1]. Lactic acid has two optical isomers, L-(+)-lactic acid and D-(−)-lactic acid. Lactic acid is classified as GRAS (generally recognized as safe) for use as a food additive, although D(-)-lactic acid can be harmful to human metabolism and result in e.g. acidosis [2]. The optical purity of lactic acid is required for the production of PLA. The properties of PLA may however be adjusted by the ratio of the L- and D-PLA in a copolymer D-form increasing the melting point of the copolymer [3]. Optically pure L- or D- lactic acid can be obtained by microbial fermentation and presently more than 95 % of industrial production of lactic acid is based on fermentation.

Production figure of 260,000 t as 100 % lactic acid for conventional (excluding PLA) markets in 2008 and forecast over 1 million ton annual production of lactic acid for conventional markets and PLA by 2020 has been presented in 2010 [4]. DuPont patented PLA already in 1954 but it took almost 50 years before first large-scale production was started. The US-based NatureWorks is the largest producer of PLA having lactic acid production capacity of 180,000 t/a. The sustainability of the PLA product Ingeo® from NatureWorks has been evaluated [5]. Greenhouse gas emissions and nonrenewable energy consumption for Ingeo from cradle to factory-gate are 1.3 kg CO2 eq./kg polymer and 42 MJ/kg polymer. These compare favorably with e.g. fossil-based PET (polyethylene terephtalate) with 3.2 kg CO2 eq./kg polymer and 80 MJ/kg polymer, respectively. There is a huge potential for biodegradable and renewable raw materials based polymers if and when the economics for these become competitive. It is estimated that altogether 140 million tons of petroleum-based synthetic polymers are produced annually [6]. It should be emphasized that also many petroleum-based synthetic polymers (e.g. polyesters) are biodegradable. However at the moment there are only three commercial synthetic polymers replacing petroleum-based ones and produced on renewable raw materials: PLA, PTT (polytetramethylene terephtalate which is partly renewable) and PHA (polyhydroxyalkanoates). Natural polymers such as starches and celluloses are biodegradable and based on renewable raw materials, but their applications are limited by their properties. Reliance Life Sciences is producing copolymers of PLA and glycolic acid mainly for high-value medical applications. Lactic acid in this case is produced by bacterial fermentation.

The price of PLA is ca. 2.2 $/kg, the target being half of that [7]. This means that the price of lactic acid in captive use should be less than 0.8 $/kg. A major cost factor is the raw material used in fermentation medium. This is especially the case with fastidious lactic acid bacteria. Processes based on cheap polymeric waste and side stream materials are indeed widely studied. So far research on alternative fermentation modes and reactor systems has been mainly academic. PLA production requires both optically and chemically pure lactic acid. Optical purity can be guaranteed with several microbial strains under optimized fermentation conditions. Chemical purity is mainly dependent on the constituents in the fermentation medium especially when cheap materials are being used. Contrary to many other fermentation products lactic acid yield on monosaccharides is usually very high (> 90 %) the main impurity being the cell mass itself, which is easily separated from the product. The key economic drivers in the fermentative production of lactic acid are optimization of the production medium, high product yields, productivity, and the concentration of products formed, which influences the down-stream processing costs [8].

Lactic acid bacteria (LAB) are a group of Gram-positive bacteria belonging to genera Aerococcus, Alloiococcus, Atopobium, Bifidobacterium, Carnobacterium, Enterococcus, Lactobacillus (Lb.), Lactococcus (L.), Leuconostoc (Leuc.), Oenococcus, Pediococcus, Streptococcus (S.), Tetragenococcus, Vagococcus and Weissella (W.). LAB are non-sporulating rods or cocci which produce lactic acid as the main fermentation product under suitable substrates. LAB are oxidase and benzidine negative, lack cytochromes, and do not reduce nitrates to nitrite [9]. Most of the LAB are anaerobic, but some of them can shift to oxygen-dependent metabolism in aerobic conditions [10,11]. Lactic acid bacteria have complex nutrient requirements, including specific minerals, B vitamins, several amino acids, and purine and pyrimidine bases.

LAB ferment sugars via homo-, hetero-, or mixed acid fermentation. Homofermentative LAB produce lactic acid as main product from sugars, while hetero- or mixed acid fermentations produce also ethanol and/or acetic acid, formic acid and carbon dioxide. Although it is a common practice to divide LAB into homo- and heterofermentative strains, the division is not that straightforward as the actual metabolism is dependent on both the nature of the C/energy substrate (e.g. hexose vs. pentose sugars) and fermentation conditions (e.g. growth rate and availability of the C/energy source). LAB used for lactic acid production are used to be classified as homofermentative (Lactococcus, Enterococcus, Streptococcus and some lactobacilli) as their hexose metabolism under non-limiting conditions is entirely via Embden-Meyerhof pathway to pyruvate which is then used to regenerate the reducing power (NADH) in the lactate dehydrogenase (LDH) catalyzed reaction to lactic acid. However at slow growth rate and low glycolytic flux mixed acid fermentation may take place and acetic acid, formic acid and ethanol are formed in addition to lactic acid [12]. The key enzyme in this metabolic shift e.g. in L. lactis is claimed to be pyruvate-formate lyase (PFL) [13]. There are two types of LDH for both enantiomers D-LDH and L-LDH. In addition some species have a racemase enzyme catalyzing the reaction between the two enantiomers. Thus enantiomerically pure lactic acid is produced by species with only one type of LDH and no racemase. A comprehensive list of different LAB strains used in lactic acid production is available elsewhere [1].

Biotechnical production of lactic acid may be based on several alternative micro-organisms. In addition to lactic acid bacteria filamentous fungi (e.g. Rhizopus spp.), other gram-positive bacteria (e.g. Bacillus coagulans) and metabolically engineered yeasts have been used also in industrial scale. The advantage of fungi is that they are active at and tolerate low medium pH. Low pH reduces significantly the consumption of neutralizing agent (Ca(OH)2) in the fermentation stage and subsequent formation of gypsum (CaSO4) in the product recovery stage. The advantage of filamentous fungi, Bacillus spp. and yeasts compared to lactic acid bacteria is their simple nutrient requirement in the fermentation medium. Filamentous fungi and Bacillus spp. are better suited to lignocellulosic fermentation raw materials as they are in general able to utilize pentose sugars in addition to hexoses. Anaerobic fermentation is generally speaking more feasible and this favors yeasts and lactic acid bacteria. When optimized the technical parameters such as product yield, RP and final product concentration are quite similar for each of these production organisms.

In the wide literature on lactic acid production two examples based on other than lactic acid bacteria should be taken up. The first of them presents results with a thermotolerant B. coagulans strain [14]. High lactic acid YP/S on both glucose and xylose (96 % and 88 %, respectively) were achieved at reasonable RP (2.5 g/lh) and product concentration (100 g/l). Exceptionally high levels of lactic acid (200 g/l) were produced in fed-batch fermentation. Yeasts have been metabolically engineered aiming at lactic acid production since 1990’s [15]. A recent article reported on metabolic engineering of Candida utilis having pyruvate decarboxylase deleted and a bovine L-lactate dehydrogenase expressed under the pdc promoter resulting in the production of lactic acid with high yield from glucose (95 %) and reasonable RP (4.9 g/lh) ending up with lactic acid concentration of 103.3 g/l and more than 99.9 % enantiomeric purity [16].

Heterofermentative LAB (Leuconostoc, Weissella and some lactobacilli such as Lb. brevis) utilize both hexose and pentose sugars via phosphoketolase pathway (PKP). Several LAB possess the genes for PPP as well. The different pathways are presented in Fig. 1. Heterofermentative LAB may be applied for the production of side products such as polyols (mannitol, erythritol) and ethanol or acetic acid. This is only feasible if the markets for the side products are comparable to those of lactic acid and the production more than covers the added down-stream processing costs.

Figure 1.

The main metabolic pathways in LAB. EMP: Embden-Meyerhof-Parnas pathway. PPP: pentose phosphate pathway. PKP: phosphoketolase pathway. Glu: glucose. LA: lactic acid. HAc: acetic acid. FA: formic acid. EtOH: ethanol. -P: energy-rich phosphate group. Pi: inorganic phosphate. Xyl: xylose. Xu: xylulose. 6-PG: 6-phosphogluconate. Ru: ribulose. R: ribose. Ga: glyceraldehyde. E: erythrose. Su: seduheptulose. Fru: fructose. Ac-CoA: acetyl-coenzymeA. Pyr: pyruvate. AcA: acetaldehyde. Ac-P: acetyl-phosphate. Pfl: pyruvate-formate lyase. XylA: xylose isomerase. XylB: xylulokinase. All carbohydrates are in D-form. Various metabolic end-products are presented with the dark background.


2. Future raw materials for production of lactic acid by LAB

The carboxylate platform is comprised of biological and chemical pathways that can be used in order to convert waste to bioproducts, such as lactic acid [17]. Lactic acid is a relatively cheap product, and one of the major challenges in its large-scale fermentative production is the cost of the raw material. This is the situation even in case of so called low-cost substrates [18]. Therefore, development of processes that utilize cheap raw materials at minimal costs have been under extensive studies. These substrates can be roughly classified as starch-based non-processed biomasses, lignocellulosic non-processed biomasses, and waste or side stream feedstocks. The former are nowadays generally considered as non-ideal feedstocks due to ethical reasons, and therefore they are not discussed in this review. Extensive reviews including starch-based feedstocks are available elsewhere [19]. With respect to future applications, the most likely raw materials for the lactic acid production are industrial side-streams and lignocellulosic biomasses. Recent advances in case of both raw material groups are discussed in the following.

As in other bioconversion processes, also in lactic acid production the focus of research has turned towards the use of lignocellulosic feedstocks. The major driving forces are fossil fuel deprivation and general paradigm change to bioeconomy, and the abundancy of lignocellulose materials. Generally, the effective utilization of lignocellulosic biomass for biochemical processes is limited due to seasonal availability, scattered distributions and high logistics cost [20]. The fermentation of lignocellulosic biomasses can also be hampered by inefficient pretreatment, high enzyme costs and end-product inhibition, formation of unwanted by-products under metabolism of pentoses, and carbon catabolite repression caused by the heterogeneous substrates. These challenges are further discussed in a recent review [8].

Paper industry residues and recycled paper products include various possible feedstocks for lactic acid production, which are together with agroindustrial residues discussed in a recent review [21]. Due to economical and ecological reasons, an intensive research interest is currently devoted to complex industrial by-products. In this field the advances presented during the past five years include the utilization of cellulosic biosludges from a Kraft pulp mill [22,23], and recycled paper sludge [24]. In both cases a nutrient supplement has increased the lactic acid productivity. LAB could be used for the bioconversion of hemicellulose fractions, e.g. from alfafa processing [25] to lactic acid. Direct conversion of xylan to lactic acid by LAB is already possible by use of genetically modified strains [26].

Food industry residues comprise a large variety of different biomasses and sludges that can be roughly categorized to agricultural wastes and food production wastes. Since the use of agricultural residues for lactic acid production is summarized in a recent review [21], it is not futher discussed here. Food production residues have been tested for bioconversion applications for ages, and the variety of used materials is large. Whey and other dairy industry residues are the prominent raw materials with respect to lactic acid production, reviewed in e.g. [27,28]. Whey retains about 55% of total milk nutrients, from which approximately 70% consists of lactose [29]. Availability of the lactose carbohydrate reservoir and the presence of other essential nutrients, such as proteins and phosphates, for the growth of microorganisms make whey and other cheese-making residues potent raw materials for the production of biochemicals.

Other quite often referred raw materials include brewery residues, especially spent grain [30], and winery wastes [31-35]. Additionally, there are various other proposed food industry residues that could fit to the lactic acid fermentation. The recently proposed include e.g. apple pomace [36], canned pineapple syrup [37], cashew apple juice [38], Jerusalem artichoke tubers [39], macaroni milk and rice-green pea-salad refectory wastes [40], rice residues [41], sap from palmyra and oil palms [42], and spent coffee grounds [43]. Despite of the large variety of the raw materials, the main conclusions of these studies are that the optimization and control of pH and temperature is critical for the process, and that the supplementation of low-cost substrate with e.g. inorganic salts and yeast extract is necessary or at least improves the productivity remarkably. In a recent study the use of mixed cultures of Lb. casei, Lb. helveticus, and S. thermophilus was observed to reduce the demand of supplements compared to single strain cultures [44].

The required supplements and their concentrations depend on the low-cost substrate. Drawbacks of complex supplements are their cost and extensive down-stream processing required for the purification of lactic acid from fermentation broth, especially in applications requiring high purity. Therefore, the optimization of supplement concentration is essential. Although yeast extract is often considered superior to other supplements in terms of efficiency, its major drawback is the relatively high cost, and therefore substitutive supplements have been suggested. Equal productivities may be achieved via use of cheaper alternatives, such as inorganic phosphates [45], and microbial lysates [46,47]. It is notable that the use of lysates in combination to e.g. whey proteins could cause unwanted proteolytic activity. Other options for the increased productivity include e.g. the addition of manganese, which is a constituent of lactate hydrogenase [48], whey protein hydrolyzate [49], malt combing nuts [49], corn steep liquor [50], fish hydrolyzates and other fishery by-products [51-53], hydrolyzed spent cells [54] or red lentil flour [55]. It is likely that this is one of the future trends in lactic acid production, i.e. fermentation media are optimized from mixtures of different low-cost raw materials in order to avoid the use of expensive complex supplements.

The modern biorefineries are looking into oceans in order to find new abundant and less land- or water-using biomasses for the production of commodities. Among the plenty marine biomasses, brown seaweed and especially species Laminaria japonica, a common food in Japan, has been recognized as a potential raw material for the production of platform chemicals. L. japonica is interesting due to its high carbohydrate content and fast growth. Production of lactic acid from L. japonica hydrolyzates was reported in a recent study [56]. Another potential raw material for bioconversion is shrimp shell waste, which is produced in vast amounts as a by-product of food industry. It has been reported that the production of lactic acid can be combined to the recovery of biopolymer chitin, a precursor for largely applied chitosan [57,58]. Since the recovery of chitin is traditionally done via chemical processing, the integrated process offers both economical and ecological advantage. Similar to the previous examples of other food industry residues, also the marine food processing industry generates various different side streams, such as fish waste and shells that could perhaps be combined in biochemical production.


3. Novel LAB strains

Metabolic engineering in general is applied when e.g. YP/S, RP, substrate flux through a desired pathway in growth phases or resting cells are aimed at. Metabolic engineering studies aiming at increased flux in glycolysis to lactic acid in LAB are fairly scarce. That may be explained by the fact that the metabolism of LAB is already tuned for efficient lactic acid production.

Some of these studies are listed in a review on metabolic engineering for lactic acid production [59]. The overexpression of L-LDH in Lb. plantarum can result 13-fold increase in LDH activity, and still show no effect on lactic acid production [60]. It has also been shown by overexpression that glyceraldehyde-3-P dehydrogenase (GAPDH) is not limiting the glycolytic flux either in growing or resting cells of a L. lactis strain [61]. Metabolic flux and control analysis (MFA and MCA) combined with the estimation of the kinetic parameters of the enzymes of a pathway are indeed needed in systematic and systemic approach to study and optimize also such seemingly simple - there is always growth and maintenance functions involved as well - metabolic pathway as that from glucose to lactic acid in LAB. An excellent view on topic is available in a review [62], which includes several references also for LAB (e.g. [63-66]).

More straightforward work on lactic acid production has been performed to achieve high enantiomeric purity by expressing and deleting respective genes for LDH. There are several examples of these as discussed in the recent review [59], such as construction of two different strains of Lb. helveticus for optically pure L-lactic acid production [67]. These strains differed from each other at the level of L-LDH activity (53 and 93 % higher than the wild type strain). Lactic acid production in a fermentation batch was equal to that of the wild type strain. However, at low pH when the growth and production are uncoupled, the strain with higher activity produced 20 % more lactic acid compared to construct with the lower activity.

Another straightforward target for the construction of genetically modified strains is widening of the raw materials for the production of lactic acid especially to lignocellulosic biomass-based materials. There are no reports on work to produce cellulolytic enzymes in LAB. Instead several groups have tried to produce xylanase in LAB [26]. This is however focused on heterofermentative LAB as they are naturally able to utilize pentoses and especially Lb. brevis as it has been shown to have endogenous beta-xylosidase activity [68]. Another approach is based on L. lactis IO-1 strain being able to metabolize xylose both via PKP and PPP [69]. PPP provides a homolactic fermentation route for pentoses. As the molecular biology tools or protocols for this strain were not available, another strain of L. lactis was used as the host. XylRAB genes from IO-1 strain were expressed in the host. XylA and XylB encode genes for xylose isomerase and xylulokinase, respectively. XylR is a putative transcriptional activator of the XylAB operon. In addition the gene for phosphoketolase was disrupted. Such a strain construct had homolactic fermentation for xylose. The rate of xylose fermentation was further improved by overexpressing the gene for transketolase, one of the enzymes in PPP. Almost theoretical YP/S of lactic acid (1.58 vs. 1.67 mol/mol xylose) was achieved with lactic acid concentration of 50,1 g/l. Acetic acid concentration was as low as 0.3 g/l. Enantiomeric purity was very high (99.6 %). Similar approach has been applied for the production of D-lactic acid from xylose and L-arabinose [70,71].

Typical LAB fermentations are run at minimum pH of 5 – 5.5, which is much higher than the pKa-value of lactic acid (i.e. 3.8). Thus more than 90 % of the product exists as lactate. This is a major cost factor in the product recovery stage as well as the cause of high salt burden and/or gypsum formation. The tolerance to acid and low pH is difficult to explain at genetic level and thus hardly be affected by metabolic engineering methods on specific genes. A successful approach to engineer LAB strains for lower fermentation pH has been genome shuffling. E.g. populations from nitrosoguanidine (NTG) mutations and low pH acclimatization in chemostat cultivation have been used for the shuffling [72]. The resultant population grew at pH 3.8 and lowered pH by lactic acid formation down to 3.5. This is a promising result even though the population was not used with realistic sugar concentrations. Similar approach has been reported aiming at improving acid tolerance as well as RP and glucose tolerance, respectively, with Lb. rhamnosus [73,74]. NTG and UV irradiation were used for mutagenesis and lethal mutants were fused from protoplasts. The best strain of [73] lowered pH down to 3.25 and increased average RP by 60 % compared to the wild type strain. However, average RP was still moderately low (ca. 1 g/lh). Final lactic acid concentration and YP/S from glucose were 84 g/l and 82 %, respectively. In [74] higher YP/S (> 95 %) and RP (ca. 3.6 g/lh) were reached with the best strains on industrially relevant fermentation medium with 150 g/l glucose. The YP/S from 200 g/l glucose was still 90 %, but the average RP decreased to 2 g/lh. In a recent study Lb. casei mutants induced by NTG were screened in high glucose concentration (360 g/l) [75]. A mutant strain with highest osmotic tolerance produced 198.2 g/l lactic acid from 210 g/l glucose with increased RP (5.5 g/lh).


4. Novel process technologies

From fermentor design point of view lactic acid production by LAB is quite simple and conventional as the process requires no gassing, gas exchange or gas mass transfer. When the production strain and fermentation conditions are optimized for lactic acid production there is no or little formation of side products (metabolites, cellular mass, exopolysaccharides). Thus e.g. the rheology of the fermentation broth is Newtonian and very close to that of water. Power consumption is mainly for the sake of homogeneity and reduction of gradients of pH-controlling agents. The biggest challenges for process technology are to minimize osmotic effects by substrates and the product, to reach high RP and to minimize the costs and waste formation in the product recovery stage.

Typical fermentation approaches other than simple batch include repeated batch and fed-batch fermentation and continuous fermentation with cell-recycle as solutions with free cells and the use of immobilized cells in different reactor types (fixed or fluidized bed). A novel fed-batch strategy was developed recently by combining pH-control and substrate feeding [76]. The rationale behind the strategy was the linear relationship between the consumption amounts of alkali and that of substrate. Thus these two components were mixed together in the feeding liquid. This resulted in higher efficiency compared to batch fermentation, but the efficiency parameters were not especially high if compared with data from several other reports. By far the most studied method to increase the RP and/or separate cell growth from product formation is based on the immobilization of the cells. These have also been reviewed [27]. Several immobilization methods have been applied including entrapment within gels such as alginate [77,78], modified alginate [79,80], or pectate [81], adsorption on granulated DEAE-cellulose [82] or porous glass [83], and biofilm formation on solid supports [84,85]. Solid incompressible supports and carrier materials such as granulated cellulose and porous glass may be applied in any scale and in various reactor designs while gels as compressible materials suit less well for larger scale especially in fixed-bed column reactors.

Immobilized cells may be utilized in various fermentation modes and reactor designs such as repeated batch or fed-batch, continuous fermentation with cell retention or recycle, in continuous stirred tank reactors (CSTR), fixed-bed or fluidized-bed reactors. High RP (19-22 g/lh) have been achieved in a two-stage process with immobilized cells [86]. A special arrangement consisting of a CSTR for pH-control and substrate feeding and a fixed-bed reactor with immobilized cells was used in a concept with intermittent refreshing of the cells in a patent [87]. Short residence time within the column was possible because of the incompressible nature of the carrier material. Chemically pure product was achieved by using a production medium with few nutrients. Once the productivity decreased below a threshold value based on the consumption of alkali the cells were refreshed with nutrients. Incompressible carriers for cell adsorption have obvious advantages. However, new solutions to secure cell adherence on the carrier are required. This would facilitate efficient use of fluidized-bed reactors with minimal pressure losses in the reactor. Biological means for cell adherence may be one solution which could offer a further advantage to selectively keep the productive cells in the reactor.

Another approach to increase RP is high cell-density fermentation with free cells recycled by membrane separation technique. This has been in use in industrial scale for lactic acid production already in 1980’s in Finland. Several academic reports on this approach have been since published demonstrating very high RP of 26 g/lh [88], 31.5 g/lh [89] and up to 57 g/lh [90].

It should be kept in mind that RP is affected by the concentration of lactic acid. Thus not all published figures are comparable. Product inhibition may be diminished by in-situ recovery of the product. Electrodialysis [91,92], nanofiltration [93] and ion-exchange [94,95] have thus been coupled with the fermentation system.

Conventional lactic acid recovery from fermentation broth consists of cell and other solids separation, lactic acid precipitation as calcium lactate and precipitate recovery, acidification of the precipitate by sulfuric acid and the separation of the gypsum precipitate formed. The amount of gypsum is usually higher than the amount of lactic acid produced. Lately NatureWorks has reported to have reduced the formation of gypsum significantly. Probably this has been achieved by performing the fermentation at lower pH e.g. by using metabolically engineered yeast for the production of lactic acid. The amount of gypsum can be avoided by using electrodialysis for the acidification and separation of the acid and alkali formed with bipolar membranes [96]. The alkali formed may be recycled back to the fermentation. Electrodialysis has been considered too expensive technology for lactic acid recovery [97]. However, specific energy consumption of only 0.25 kWh/ kg lactic acid is presented [96]. Nanofiltration has been used as a pretreatment method to remove Mg- and Ca- and sulfate-ions and color before electrodialysis increasing significantly the capacity in electrodialysis [98]. Alternative techniques for lactic acid recovery are extraction [99] and use of ion-exchange [100,101], neither of which is a proper solution to the salt burden.


5. Conclusions

Lactic acid production in LAB has both cell mass and growth dependent portions. Typically LAB require several nutrient components for their growth increasing the fermentation and down-stream processing costs. Down-stream processing is especially important in the production of lactic acid for PLA. As RP is the a major investment factor affecting costs, the minimization of medium and product purification costs should be accompanied by methods increasing cell mass concentration without excess growth. For this several different strategies have been applied so far mainly in academia (cell immobilization, cell-recycling and cell-retention). As history shows some of these could be applicable in industrial production as well, however pilot and demonstration plant studies and some risk-taking are required.

The main C/energy source spectrum available for LAB has been widened significantly. Reports of new possible substrates are frequently published, and the utilization of industrial side streams is a growing trend. Into this direction major successes have also been achieved with metabolic engineering providing strains for efficient production of lactic acid from pentoses as well, which is to promote sustainable use of renewables.

In an ideal fermentation process product inhibition should be minimized so that high RP would be achieved even at high lactic acid concentrations resulting in feasible average productivities. For this purpose both acclimatization and mutagenesis has been applied successfully. However, it has to be considered how far can we go in respect to fermentation pH and lactic acid concentration. There are already remarkable alternatives to LAB with naturally better properties in this sense. Some success has been achieved with in-situ product recovery, but also these procedures lack experiences in any larger scale.

Conventional lactic acid production process with LAB is accompanied with the formation of large amounts of gypsum in the product recovery stage. Fermentation at lower pH diminishes this amount, but does not prevent its formation. Electrodialysis has been considered too expensive technique for the recovery of such cheap, bulk products as lactic acid. However, recent reports claim promising results with this technology. Forecasted figures for lactic acid market show up to one million tons per year. The growth would come mainly from the growth of PLA as a biodegradable polymer based on renewable raw materials. Economies of scale should decrease the production costs, but new technical approaches are also needed to reach these figures.



η % - Efficiency, i.e. the ratio of YP/S to the maximum theoretical value

D-LDH - D-lactate dehydrogenase

LAB – lactic acid bacteria

L-LDH - L-lactate dehydrogenase

NTG – Nitrosoguanidine

RP - Volumetric productivity g/l*h

SSF – Simultaneous saccharification and fermentation

PLA – poly lactic acid

PPP - Pentose phosphate pathway

PKP - Phosphoketolase pathway

YP/S – Yield of lactic acid per substrate consumed g/g

YP/X – Yield of lactic acid per cell mass g/g


  1. 1. Wee YJ, Kim JN, Ryu HW.Biotechnological production of lactic acid and its recent applications. Food Technology and Biotechnology 200644163
  2. 2. DattaR.TsaiS. P.BonsignoreP.MoonS. H.FrankJ. R.Technological and Economic-Potential of Poly(Lactic Acid) and Lactic-Acid Derivatives. Fems Microbiology Reviews 199516221
  3. 3. FukushimaK.SogoK.MiuraS.KimuraY.Production of D-lactic acid by bacterial fermentation of rice starch. Macromolecular Bioscience 200441021
  4. 4. JemJ. K.van der PolJ. VosS.Microbial lactic acid, its polymer poly(lactic acid), and their industrial applications. Microbiology Monographs 201014323
  5. 5. VinkE. T. H.DaviesS.KolstadJ. J.The eco-profile for current Ingeo® polylactide production. Industrial Biotechnology 20106212
  6. 6. AAShahHasan. F.HameedA.AhmedS.Biological degradation of plastics: A comprehensive review. Biotechnology Advances 200826246
  7. 7. Nampoothiri KM, Nair NR, John RP.An overview of the recent developments in polylactide (PLA) research. Bioresource Technology 20101018493
  8. 8. MAAbdel-RahmanTashiro. Y.SonomotoK.Lactic acid production from lignocellulose-derived sugars using lactic acid bacteria: Overview and limits. Journal of Biotechnology 2011156286
  9. 9. CarrF. J.ChillD.MaidaN.The lactic acid bacteria: A literature survey. Critical Reviews in Microbiology 200228281
  10. 10. MurphyM. G.CondonS.Correlation of Oxygen Utilization and Hydrogen-Peroxide Accumulation with Oxygen Induced Enzymes in Lactobacillus-Plantarum Cultures. Archives of Microbiology 198413844
  11. 11. SedewitzB.SchleiferK. H.GotzF.Physiological role of pyruvate oxidase in the aerobic metabolism of Lactobacillus plantarum. Journal of Bacteriology 1984160462
  12. 12. ZaunmullerT.EichertM.RichterH.UndenG.Variations in the energy metabolism of biotechnologically relevant heterofermentative lactic acid bacteria during growth on sugars and organic acids. Applied Microbiology and Biotechnology 200672421
  13. 13. MelchiorsenC. R.JokumsenK. V.VilladsenJ.IsraelsenH.ArnauJ.The level of pyruvate-formate lyase controls the shift from homolactic to mixed-acid product formation in Lactococcus lactis. Applied Microbiology and Biotechnology 200258338
  14. 14. Ou MS, Ingram LO, Shanmugam KT.l(+)-Lactic acid production from non-food carbohydrates by thermotolerant Bacillus coagulans. Journal of Industrial Microbiology & Biotechnology 201138599
  15. 15. PorroD.BrambillaL.BMRanziMartegani. E.AlberghinaL.Development of Metabolically Engineered Saccharomyces-Cerevisiae Cells for the Production of Lactic-Acid. Biotechnology Progress 199511294
  16. 16. IkushimaS.FujiiT.KobayashiO.YoshidaS.YoshidaA.Genetic Engineering of Candida utilis Yeast for Efficient Production of L-Lactic Acid. Bioscience Biotechnology and Biochemistry 2009731818
  17. 17. Agler MT, Wrenn BA, Zinder SH, Angenent LT.Waste to bioproduct conversion with undefined mixed cultures: the carboxylate platform. Trends in Biotechnology 20112970
  18. 18. Yadav AK, Chaudhari AB, Kothari RM.Bioconversion of renewable resources into lactic acid: an industrial view. Critical Reviews in Biotechnology 2011311
  19. 19. JohnR. P.NampoothiriK. M.PandeyA.Fermentative production of lactic acid from biomass: an overview on process developments and future perspectives. Applied Microbiology and Biotechnology 200774524
  20. 20. LinY.TanakaS.Ethanol fermentation from biomass resources: current state and prospects. Applied Microbiology and Biotechnology 200669627
  21. 21. AlonsoJ. L.DominguezH.GarroteG.MJGonzalez-MunozGullon. B.MoureA.SantosV.VilaC.YanezR.Biorefinery processes for the integral valorization of agroindustrial and forestal wastes. Cyta-Journal of Food 20119282
  22. 22. RomaniA.YanezR.GarroteG.AlonsoJ. L. S. S. F.productionof.lacticacid.fromcellulosic.biosludgesBioresource Technology 2008994247
  23. 23. RomaniA.YanezR.GarroteG.AlonsoJ. L.ParajoJ. C.Sugar production from cellulosic biosludges generated in a water treatment plant of a Kraft pulp mill. Biochemical Engineering Journal 200737319
  24. 24. MarquesS.SantosJ. A. L.GirioF. M.RoseiroJ. C.Lactic acid production from recycled paper sludge by simultaneous saccharification and fermentation. Biochemical Engineering Journal 200841210
  25. 25. Sreenath HK, Moldes AB, Koegel RG, Straub RJ.Lactic acid production by simultaneous saccharification and fermentation of alfalfa fiber. Journal of Bioscience and Bioengineering 200192518
  26. 26. Hu CY, Chi DJ, Chen SS, Chen YC.The direct conversion of xylan to lactic acid by Lactobacillus brevis transformed with a xylanase gene. Green Chemistry 2011131729
  27. 27. KossevaM. R.PanesarP. S.KaurG.KennedyJ. F.Use of immobilised biocatalysts in the processing of cheese whey. International Journal of Biological Macromolecules 200945437
  28. 28. PanesarP. S.KennedyJ. F.GandhiD. N.BunkoK.Bioutilisation of whey for lactic acid production. Food Chemistry 20071051
  29. 29. JelenP.Wheyprocessing.InEncyclopedia.ofdairy.sciencesH.H.Roginski, J.W.Fuquay, P.F.Fox, (Eds.), Academic Press: London, 200327392751
  30. 30. AliyuS.BalaM.Brewer’sspent.grainA.reviewof.itspotentials.applicationsAfrican Journal of Biotechnology 201110324
  31. 31. Devesa-ReyR.VecinoX.Varela-AlendeJ. L.BarralM. T.CruzJ. M.MoldesA. B.Valorization of winery waste vs. the costs of not recycling. Waste Management 2011312327
  32. 32. BustosG.MoldesA. B.CruzJ. M.DominguezJ. M.Production of lactic acid from vine-trimming wastes and viticulture lees using a simultaneous saccharification fermentation method. Journal of the Science of Food and Agriculture 200585466
  33. 33. BustosG.MoldesA. B.CruzJ. M.DominguezJ. M.Production of fermentable media from vine-trimming wastes and bioconversion into lactic acid by Lactobacillus pentosus. Journal of the Science of Food and Agriculture 2004842105
  34. 34. BustosG.MoldesA. B.CruzJ. M.DominguezJ. M.Formulation of low-cost fermentative media for lactic acid production with Lactobacillus rhamnosus using vinification lees as nutrients. Journal of Agricultural and Food Chemistry 200452801
  35. 35. la TorreN.MoldesA. B.CruzJ. M.DominguezJ. M.Revalorization of hemicellulosic trimming vine shoots hydrolyzates trough continuous production of lactic acid and biosurfactants by L-pentosus. Journal of Food Engineering 200778405
  36. 36. GullonB.YanezR.AlonsoJ. L.ParajoJ. C.L-lacticacid.production from apple pomace by sequential hydrolysis and fermentation. Bioresource Technology 200899308
  37. 37. NakanishiK.UenoT.SatoS.YoshiS. L.L-Lactic Acid Production from Canned Pineapple Syrup by Rapid Sucrose Catabolizing Lactobacillus paracasei NRIC 0765. Food Science and Technology Research 201016239
  38. 38. HonoratoT. L.RabeloM. C.GoncalvesL. R. B.PintoG. A. S.RodriguesS.Fermentation of cashew apple juice to produce high added value products. World Journal of Microbiology & Biotechnology 2007231409
  39. 39. GeX. Y.QianH.ZhangW. G.Enhancement of L-Lactic Acid Production in Lactobacillus casei from Jerusalem Artichoke Tubers by Kinetic Optimization and Citrate Metabolism. Journal of Microbiology and Biotechnology 201020101
  40. 40. OmayD.GuvenilirY.Lactic Acid Fermantation from Refectory Waste. Ekoloji 20112042
  41. 41. LuZ. D.LuM. B.HeF.YuL. J.An economical approach for D-lactic acid production utilizing unpolished rice from aging paddy as major nutrient source. Bioresource Technology 20091002026
  42. 42. ChooklinS.KaewsichanL.KaewsichanL.Potential use of Lactobacillus casei TISTR 1500 for the bioconversion from palmyra sap and oil palm sap to lactic acid. Electronic Journal of Biotechnology 2011
  43. 43. MussattoS. I.MachadoE. M. S.MartinsS.TeixeiraJ. A.ProductionComposition.Applicationof.CoffeeItsIndustrial.ResiduesFood and Bioprocess Technology 20114661
  44. 44. SecchiN.GiuntaD.PrettiL.GarciaM. R.RoggioT.MannazzuI.CatzedduP.Bioconversion of ovine scotta into lactic acid with pure and mixed cultures of lactic acid bacteria. Journal of Industrial Microbiology & Biotechnology 201239175
  45. 45. AmraneA.Effect of inorganic phosphate on lactate production by Lactobacillus helveticus grown on supplemented whey permeate. Journal of Chemical Technology and Biotechnology 200075223
  46. 46. AmraneA.Evaluation of lactic acid bacteria autolysate for the supplementation of lactic acid bacteria fermentation. World Journal of Microbiology & Biotechnology 200016207
  47. 47. CoelhoL. LimaC. J. B.BernardoM. P.ContieroJ.d(-d(-)-Lactic Acid Production by Leuconostoc mesenteroides B512 Using Different Carbon and Nitrogen Sources. Applied Biochemistry and Biotechnology 20111641160
  48. 48. FitzpatrickJ. J.AhrensM.SmithS.Effect of manganese on Lactobacillus casei fermentation to produce lactic acid from whey permeate. Process Biochemistry 200136671
  49. 49. FitzpatrickJ. J.O’KeeffeU.Influence of whey protein hydrolysate addition to whey permeate batch fermentations for producing lactic acid. Process Biochemistry 200137183
  50. 50. Kim HO, Wee YJ, Kim JN, Yun JS, Ryu HW.Production of lactic acid from cheese whey by batch and repeated batch cultures of Lactobacillus sp RKY2. Applied Biochemistry and Biotechnology 2006131694
  51. 51. GaoM. T.HirataM.ToorisakaE.HanoT.Acid-hydrolysis of fish wastes for lactic acid fermentation. Bioresource Technology 2006972414
  52. 52. BeaulieuL.DesbiensM.ThibodeauJ.ThibaultS.Pelagic fish hydrolysates as peptones for bacterial culture media. Canadian Journal of Microbiology 2009551240
  53. 53. VazquezJ. A.MontemayorM. I.FraguasJ.MAMuradoHigh production of hyaluronic and lactic acids by Streptococcus zooepidemicus in fed-batch culture using commercial and marine peptones from fishing by-products. Biochemical Engineering Journal 200944125
  54. 54. GaoM. T.HirataM.ToorisakaE.HanoT.Study on acid-hydrolysis of spent cells for lactic acid fermentation. Biochemical Engineering Journal 20062887
  55. 55. AltafM.NaveenaB. J.ReddyG.Use of inexpensive nitrogen sources and starch for L(+) lactic acid production in anaerobic submerged fermentation. Bioresource Technology 200798498
  56. 56. JangS.ShiraiY.UchidaM.WakisakaM.Production of L(+)-Lactic Acid from Mixed Acid and Alkali Hydrolysate of Brown Seaweed. Food Science and Technology Research 201117155
  57. 57. AdourL.ArbiaW.AmraneA.MameriN.Combined use of waste materials- recovery of chitin from shrimp shells by lactic acid fermentation supplemented with date juice waste or glucose. Journal of Chemical Technology and Biotechnology 2008831664
  58. 58. HealyM.GreenA.HealyA.Bioprocessing of marine crustacean shell waste. Acta Biotechnologica 200323151
  59. 59. SinghS. K.AhmedS. U.PandeyA.Metabolic engineering approaches for lactic acid production. Process Biochemistry 200641991
  60. 60. FerainT.GarmynD.BernardN.HolsP.DelcourJ.Lactobacillus-PlantarumLdhl.Gene-OverexpressionDeletionJournal of Bacteriology 1994176596
  61. 61. SolemC.KoebmannB. J.JensenP. R.Glyceraldehyde-3phosphate dehydrogenase has no control over glycolytic flux in Lactococcus lactis MG1363. Journal of Bacteriology 2003
  62. 62. TeusinkB.SmidE. J.Modelling strategies for the industrial exploitation of lactic acid bacteria. Nature Reviews Microbiology 2006446
  63. 63. Bai DM, Zhao XM, Li XG, Xu SM.Strain improvement and metabolic flux analysis in the wild-type and a mutant Lactobacillus lactis strain for L(+)-lactic acid production. Biotechnology and Bioengineering 200488681
  64. 64. NevesA. R.RamosA.NunesM. VosW. M.AlmeidaJ.SantosH.In vivo nuclear magnetic resonance studies of glycolytic kinetics in Lactococcus lactis. Biotechnology and Bioengineering 199964200
  65. 65. EvenS.LindleyN. D.Cocaign-BousquetM.Transcriptionaltranslational.metabolicregulation.ofglycolysis.inLactococcus.lactissubsp.cremoris MG 1363 grown in continuous acidic cultures. Microbiology-Sgm 20031491935
  66. 66. Cocaign-BousquetM.EvenS.LindleyN. D.LoubiereP.Anaerobic sugar catabolism in Lactococcus lactis: genetic regulation and enzyme control over pathway flux. Applied Microbiology and Biotechnology 20026024
  67. 67. Kylä-NikkiläK.HujanenM.LeisolaM.PalvaA.Metabolic engineering of Lactobacillus helveticus CNRZ32 for production of pure L-(+)-lactic acid. Applied and Environmental Microbiology 2000663835
  68. 68. GardeA.JonssonG.ASSchmidtAhring. B. K.Lactic acid production from wheat straw hemicellulose hydrolysate by Lactobacillus pentosus and Lactobacillus brevis. Bioresource Technology 200281217
  69. 69. ShinkawaS.OkanoK.YoshidaS.TanakaT.OginoC.FukudaH.KondoA.Improved homo l-lactic acid fermentation from xylose by abolishment of the phosphoketolase pathway and enhancement of the pentose phosphate pathway in genetically modified xylose-assimilating Lactococcus lactis. Applied Microbiology and Biotechnology 2011911537
  70. 70. OkanoK.YoshidaS.YamadaR.TanakaT.OginoC.FukudaH.KondoA.Improved Production of Homo-D-Lactic Acid via Xylose Fermentation by Introduction of Xylose Assimilation Genes and Redirection of the Phosphoketolase Pathway to the Pentose Phosphate Pathway in L-Lactate Dehydrogenase Gene-Deficient Lactobacillus plantarum. Applied and Environmental Microbiology 2009757858
  71. 71. OkanoK.YoshidaS.TanakaT.OginoC.FukudaH.KondoA.Homo-DHomo-D-Lactic Acid Fermentation from Arabinose by Redirection of the Phosphoketolase Pathway to the Pentose Phosphate Pathway in L-Lactate Dehydrogenase Gene-Deficient Lactobacillus plantarum. Applied and Environmental Microbiology 2009755175
  72. 72. PatnaikR.LouieS.GavrilovicV.PerryK.StemmerW. P. C.RyanC. M.del CardayreS.Genome shuffling of Lactobacillus for improved acid tolerance. Nature Biotechnology 200220707
  73. 73. WangY. H.LiY.PeiX. L.YuL.FengY.Genome-shuffling improved acid tolerance and L-lactic acid volumetric productivity in Lactobacillus rhamnosus. Journal of Biotechnology 2007129510
  74. 74. YuL.PeiX.LeiT.WangY.FengY.Genome shuffling enhanced L-lactic acid production by improving glucose tolerance of Lactobacillus rhamnosus. Journal of Biotechnology 2008134154
  75. 75. GeX. Y.YuanJ. A.QinH.ZhangW. G.Improvement of L-lactic acid production by osmotic-tolerant mutant of Lactobacillus casei at high temperature. Applied Microbiology and Biotechnology 20118973
  76. 76. ZhangY.CongW.ShiS. Y.Application of a pH Feedback-Controlled Substrate Feeding Method in Lactic Acid Production. Applied Biochemistry and Biotechnology 20101622149
  77. 77. IdrisA.SuzanaW.Effect of sodium alginate concentration, bead diameter, initial pH and temperature on lactic acid production from pineapple waste using immobilized Lactobacillus delbrueckii. Process Biochemistry 2006411117
  78. 78. Shen XL, Xia LM.Lactic acid production from cellulosic waste by immobilized cells of Lactobacillus delbrueckii. World Journal of Microbiology & Biotechnology 2006221109
  79. 79. Rao CS, Prakasham RS, Rao AB, Yadav JS. Production of L(+) lactic acid by Lactobacillus delbrueckii immobilized in functionalized alginate matrices.World Journal of Microbiology & Biotechnology 2008241411
  80. 80. Rao CS, Prakasham RS, Rao AB, Yadav JS.Functionalized alginate as immobilization matrix in enantioselective L (+) lactic acid production by Lactobacillus delbrucekii. Applied Biochemistry and Biotechnology 2008149219
  81. 81. Panesar PS, Kennedy JF, Knill CJ, Kosseva MR.Applicability of pectate-entrapped Lactobacillus casei cells for L(+) lactic acid production from whey. Applied Microbiology and Biotechnology 20077435
  82. 82. LommiH.SwinkelsW.ViljavaT.HammondR.Bioreactor with immobilized lactic acid bacteria and the use thereof. WO patent 94/12614. 1994
  83. 83. SenthuranA.SenthuranV.MattiassonB.KaulR.Lactic acid fermentation in a recycle batch reactor using immobilized Lactobacillus casei. Biotechnology and Bioengineering 199755841
  84. 84. DagherS. F.RagoutA. L.SinerizF.Bruno-BarcenaJ. M.Cell Immobilization for Production of Lactic Acid: Biofilms Do It Naturally. Advances in Applied Microbiology, 712010
  85. 85. QureshiN.BAAnnousEzeji. T. C.KarcherP.MaddoxI. S.Biofilm reactors for industrial bioconversion processes: employing potential of enhanced reaction rates. Microbial Cell Factories 2005
  86. 86. SchepersA. W.ThibaultJ.LacroixC.Continuous lactic acid production in whey permeate/yeast extract medium with immobilized Lactobacillus helveticus in a two-stage process: Model and experiments. Enzyme and Microbial Technology 200638324
  87. 87. ViljavaT.KoivikkoH.Method for preparing pure lactic acid. US patent 5,932,455. 1999
  88. 88. RichterK.NottelmannS.An empiric steady state model of lactate production in continuous fermentation with total cell retention. Engineering in Life Sciences 20044426
  89. 89. XuG. Q.ChuJ.WangY. H.ZhuangY. P.ZhangS. L.PengH. Q.Development of a continuous cell-recycle fermentation system for production of lactic acid by Lactobacillus paracasei. Process Biochemistry 2006412458
  90. 90. KwonS.YooI. K.LeeW. G.ChangH. N.ChangY. K.High-rate continuous production of lactic acid by Lactobacillus rhamnosus in a two-stage membrane cell-recycle bioreactor. Biotechnology and Bioengineering 20017325
  91. 91. GaoM. T.HirataM.KoideM.TakanashiH.HanoT.Production of L-lactic acid by electrodialysis fermentation (EDF). Process Biochemistry 2004391903
  92. 92. Min-tianG.KoideM.GotouR.TakanashiH.HirataM.HanoT.Development of a continuous electrodialysis fermentation system for production of lactic acid by Lactobacillus rhamnosus. Process Biochemistry 2005401033
  93. 93. JeantetR.MauboisJ. L.BoyavalP.Semicontinuous production of lactic acid in a bioreactor coupled with nanofiltration membranes. Enzyme and Microbial Technology 199619614
  94. 94. MonteagudoJ. M.AldaveroM.Production of L-lactic acid by Lactobacillus delbrueckii in chemostat culture using an ion exchange resins system. Journal of Chemical Technology and Biotechnology 199974627
  95. 95. SenthuranA.SenthuranV.Hatti-KaulR.MattiassonB.Lactate production in an integrated process configuration: reducing cell adsorption by shielding of adsorbent. Applied Microbiology and Biotechnology 200465658
  96. 96. Wee YJ, Yun JS, Lee YY, Zeng AP, Ryu HW.Recovery of lactic acid by repeated batch electrodialysis and lactic acid production using electrodialysis wastewater. Journal of Bioscience and Bioengineering 200599104
  97. 97. AkerbergC.ZacchiG.An economic evaluation of the fermentative production of lactic acid from wheat flour. Bioresource Technology 200075119
  98. 98. BouchouxA.Roux-deBalmann. H.LutinF.Investigation of nanofiltration as a purification step for lactic acid production processes based on conventional and bipolar electrodialysis operations. Separation and Purification Technology 200652266
  99. 99. YankovD.MolinierJ.KyuchoukovG.AlbetJ.MalmaryG.Improvement of the lactic acid extraction. Extraction from aqueous solutions and simulated fermentation broth by means of mixed extractant and TOA, partially loaded with HCl. Chemical and Biochemical Engineering Quarterly 20051917
  100. 100. GullonB.AlonsoJ. L.ParajoJ. C.IonIon-Exchange Processing of Fermentation Media Containing Lactic Acid and Oligomeric Saccharides. Industrial & Engineering Chemistry Research 2010493741
  101. 101. Moldes AB, Alonso JL, Parajo JC.Recovery of lactic acid from simultaneous saccharification and fermentation media using anion exchange resins. Bioprocess and Biosystems Engineering 200325357

Written By

Sanna Taskila and Heikki Ojamo

Submitted: November 9th, 2011 Published: January 30th, 2013