Antiyeast activities of crude saponin fractions (MIC μg/ml)
1. Introduction
Saponins are a class of natural products which are structurally constructed of aglycone (triterpene or steroid) and sugars (hexose and/or uronic acid). The name ‘saponin’ comes from soap as its containing plants agitated in water form soapy lather. Saponins are widely distributed in many plants and are relatively widespread in our foodstuffs and herbal preparations. Saponins traditionally used as a natural detergent. In addition to this physical property, plant-derived triterpenoid and steroidal saponins have historically received a number of industrial and commercial applications ranging from their use as sources of raw materials for the production of steroid hormones in the pharmaceutical industry, to their use as food additives and as ingredients in photographic emulsions, fire extinguishers and other industrial applications which take advantage of their generally non-ionic surfactant properties [1-3]. They also exhibit a variety of biological activities, and have been investigated toward the development of new natural medicines and prove the efficacy of traditional herbal medicines [4]. Other interesting biological applications for various specific saponins include their uses as anti-inflammatory [5], hypocholesterolemic [6] and immune-stimulating [7] whose properties are widely recognized and commercially utilized.
As to the application of saponins to foods and cosmetics, it is indispensable that sufficient amounts of plant resources are available, and that the content of saponins must be high. Furthermore, a plant must have a long history of human use as foodstuffs or ingredients of cosmetics, and their safety should be officially guaranteed.
The saponins of Quillaja bark and licorice root are widely utilized in the world. The
Nearly 50,000 tons of licorice roots (
It is known that the deterioration of cooked foods is caused mainly by yeast, and that many skin diseases are due to infection by dermatophytic fungi and yeasts. In an expansion of utilization of saponins in foods and cosmetics, we have examined antifungal and antiyeast saponins.
2. Screening of antiyeast saponins
Crude saponin fractions from several plants were subjected to an antiyeast screening test using
Inhibitory activity against each yeast was determined using agar dilution method. The inhibitory activity of the samples was assessed as the minimum inhibitory concentration (MIC), the lowest concentration tested at which no growth was observed.
Table 1 shows the screening results of antiyeast activity tests of crude saponin mixtures from several plants. The saponin fraction from licorice root, quillaja bark, gypsophila root and soy bean seed showed no activity (MIC:>1000μg/ml) and that of hedera leaf, marronier seed, ginseng root, camellia seed, saponaria rhizome and tea seed showed a weak activity (MIC:500∼1000μg/ml), wheras crude saponin fraction from pericarps of
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Licorice root | >1000 | >1000 | Ginseng root | 1000 | 1000 |
Quillaja bark | >1000 | >1000 | Camellia seed | 1000 | 1000 |
Gypsophila root | >1000 | >1000 | Saponaria rhizome | NT | 1000 |
Hedera leaf | 1000 | 1000 | Tea seed | 500 | 500 |
Soy been seed | NT | >1000 | Yucca stem | NT | 250 |
Marronier seed | 1000 | 1000 | Sapindus pericarp | 250 | 250 |
Table 1.
3. Sapindus pericarps
Addition of an antifungal and antiyeast ingredient to cosmetics is desirable for the protection of skin against, and prevention of, dandruff generation, dermatomycosis and cutaneous candidiasis.
Significant antiyeast activity was observed for the crude saponin fraction from the pericarps of
4. Antifungal and antiyeast oleanane-saponins of Sapindus pericarps
The percarps were extracted with hot 90% MeOH. A suspension of the MeOH-extract in H2O was chromatographed on a column of highly porous polymer (Diaion H-20) eluting with H2O and 50%- and 85%-MeOH, successively. 85%-MeOH eluate gave a saponin-mixture (mono- and bis-desmosides, SP-mix). Hederagenin (1) was obtained from SP-mix by usual acid hydrolysis. Saponins 2-7 were isolated from SP-mix, such as monodesmosides: saponin A (2), sapindoside B (3), saponin C (4), sapindoside A (5), mukurozi-saponin E1 (6) etc. and bisdesmosides: mukurozi-saponin Y1 (7) etc. [10]. The structures of these saponins are shown in Figure 1.
Antidermatophytic activities of these saponins are shown in Table 2. All the monodesmosides exhibited strong growth inhibition. It is noteworthy that activity of sapindoside A is almost as strong as that of griseofulvin, the well-known antidermatophytic antibiotic. Griseofulvin does not show inhibitory activity against a pathogenic yeast,
It was found that while purified monodesmosides of pericarps are sparingly soluble in water, their solubility was greatly increased in the presence of bisdesmosides [10]. These phenomena are important for the biological activities of the pericarps.
5. Structure-antifungal activity relationship
Figure 1 showed antidermatophytic activity against
It was disclosed that for growth inhibition, the presence of free 28-COOH, 23-OH and 3-
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25 | 25 | 25 | 12.5 | 50 |
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6.25 | 6.25 | 6.25 | 3.13 | 12.5 |
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6.25 | 6.25 | 3.13 | 3.13 | 12.5 |
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6.25 | 6.25 | 6.25 | 3.13 | 25 |
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3.13 | 1.56 | 3.13 | 1.56 | 12.5 |
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6.25 | 6.25 | 6.25 | 3.13 | 12.5 |
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>100 | >100 | >100 | >100 | >100 |
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>100 | >100 | >100 | >100 | >100 |
griseofulvin* | 3.13 | 1.56 | 0.78 | 1.56 | >100 |
* positive control |
Table 2.
Antimicrobial activities of saponins and saponin mixture (
6. Antimicrobial activity of the saponin fraction of Sapindus pericarps
For commercial utilization as ingredient in cosmetics, the saponin fraction was prepared as follows. The methanolic extract was subjected to chromatography on Diaion HP-20. After removal of other water-soluble constituents by elution with water and then 50% of MeOH, the saponin fraction was obtained by elution with 80% MeOH.
The saponin fraction showed moderate antibacterial activity against Gram-positive bacteria, while no activity was obserbed against Gram-negative bacteria (Table 3).
A summarized in Table 4, the saponin fraction exhibited growth inhibition against food deteriorating yeasts,

Figure 1.
Structure and antifungal activities of saponins on

Figure 2.
Structure-antimicrobial activity relationship of oleanane-type saponin analogues
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400 |
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>400 | |
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400 |
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>400 | |
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400 |
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>400 | |
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400 |
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>400 |
Table 3.
Antibacterial activity of saponin mixture (SP-mix)
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100 |
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100 |
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50 |
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50 |
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200 |
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>400 |
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>400 |
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>400 |
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100 |
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>400 |
Table 4.
Antiyeast and antifungal activity of saponin Mixture (

Figure 3.
Saponins from Mohave yucca
7. Sapindus saponin fraction as an antidermatophytic ingredient in cosmetics
It is difficult to use
8. Mohave Yucca (Yucca schidigera)
9. Antiyeast and antifungal spirostanoid saponins from Mohave yucca
The presence of steroidal saponins in this plant has been reported previously [21,22]. As to the saponin constituents of this plant, a monodesmoside named YS-1 is isolated and identified as in [23]. We have conducted the isolation and identification of individual saponins that had not been achieved prior to this study [24,25].
The EtOH extract of this plant was subjected to colomn chromatography on highly porous polymer, Diaion HP-20, which is styrene-divinylbenzene polymer. After successive elution with water and 60% and 80% MeOH, a saponin fraction which showed significant antiyeast activity against
Figure 3 shows the structure of all of these saponins and their sapogenins. The antiyeast activities of each saponin from
Those saponins having a branched-chain trisaccharide moiety without any oxygen functionalities at C-2 and –12 exhibited potent antiyeast activities, while saponins with 2β-hydroxyl (5,6,13, and 14) or 12-keto (4 and 12) groups showed very weak or no activity. A saponin (11) with a disaccharide moiety exhibited relatively low activities. The aglycons showed no antiyeast activity.
10. Antimicrobial activity of the saponin fraction
For the commercial utilization of Mohave yucca, the antimicrobial activity of the saponin fraction which was obtained by column chromatography of the extract on Diaion HP-20 (
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3.13 | 6.25 | 3.13 | 3.13 | 12.5 | 6.25 |
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12.5 | 12.5 | 3.13 | 3.13 | >100 | >100 |
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12.5 | 12.5 | 6.25 | 3.13 | >100 | >100 |
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>100 | >100 | >100 | >100 | >100 | >100 |
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100 | 100 | >100 | 100 | >100 | >100 |
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>100 | >100 | >100 | >100 | >100 | >100 |
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6.25 | 50 | 3.13 | 3.13 | >100 | 6.25 |
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25 | >100 | 3.13 | 3.13 | >100 | 50 |
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6.25 | >100 | 3.13 | 12.5 | >100 | 6.25 |
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12.5 | 25 | 3.13 | 6.25 | 50 | 6.25 |
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12.5 | 12.5 | 6.25 | 3.13 | >100 | >100 |
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100 | >100 | 100 | >100 | >100 | >100 |
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100 | >100 | >100 | >100 | >100 | 100 |
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>100 | >100 | >100 | 100 | >100 | >100 |
Table 5.
Antiyeast activity of
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>1,000 | ||||
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1,000 |
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1,000 |
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>1,000 | |||
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>1,000 |
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>1,000 | |||
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>1,000 | ||||
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1,000 |
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>1,000 | |||
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1,000 | |||||
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>1,000 |
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1,000 |
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>1,000 | |||
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1,000 |
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>1,000 | |||
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1,000 |
Table 6.
Antibacterial acrivity of yucca saponin fraction
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62.5 | |||||||
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62.5 |
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31.3 |
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31.3 | ||||||
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62.5 |
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62.5 | ||||||
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31.3 |
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31.3 | ||||||
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31.3 |
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125 | ||||||
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31.3 |
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125 |
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31.3 | ||||||
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31.3 | |||||||
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31.3 |
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31.3 | ||||||
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31.3 | ||||||||
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>1,000 |
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>1,000 | ||||||
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>1,000 |
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>1,000 | ||||||
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125 |
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15.6 | ||||||
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>1,000 |
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>1,000 |
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>1,000 | ||||||
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>1,000 |
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>1,000 | ||||||
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>1,000 | ||||||||
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31.3 | |||||||
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15.6 |
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31.3 | ||||||
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31.3 |
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62.5 |
Table 7.
Antiyeast and antifungal acrivity of yucca saponin fraction
* food deteriorating yeast ** film-forming yeast in soy sauce
The antiyeast and antifungal activities are summarized in Table 7. The saponin fraction exhibited potent antiyeast activity. Infection of boiled rice such as “sushi” and “musubi” with
The saponin fraction showed less activity against common fungi, while it significantly inhibited the growth of dermatophytic yeast and fungi.
Potassium sorbate has been utilized in foods as a preservative. Its antiyeast activity depends upon pH. Between pH 5.0 – 3.0, potassium sorbate completely inhibited the growth of yeast at the concentration of 0.05%, while at less acidic pH (near neutral), the activity decreased remarkably. In contrast to this, such pH dependence was not observed for the yucca saponin fraction. In the range of pH 6.3 – 3.0, it entirely inhibited the growth of yeasts at the concentration of 0.03%.
11. Effects of several culture conditions against antimicrobial activity of yucca extract
The inhibitory effects of yucca extract on the growth of the yeasts isolated from ume-zuke, a salted Japanese apricot fruit product were investigated with (2% or 5%) or without sodium chloride (Table 8). From the results of MICs of yucca extract without sodium chloride, the genera
The inhibitory effect was enhanced and showed a broad antiyeast spectrum when yucca extract was used in combination with sodium chloride.
Table 9 shows the effects of several cultural conditions against antiyeast activity of yucca extract. The antiyeast activity of yucca extract was strengthened under the condition of chemical and physical conditions, low pH, alcohol, heating and high OP. While the high-polymer substances, such as polysaccharides and protein reduced antiyeast activity of yucca extract. It is interested that antiyeast activity of yucca extract was inhibited by free unsaturated fatty acids, palmitoleic acid, oleic acid and linoleic acid. On the other hand, saturated fatty acids, palmitic acid and stearic acid and oils composed of unsaturated fatty acids, olive oil, soybean oil and egg lecithin had no effect on the antiyeast activity of yucca extract.
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1000 | 500 | 250 |
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500 | 250 | 250 | |
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>2000 | 1000 | 500 |
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250 | 250 | 250 | |
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>2000 | 2000 | 500 |
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250 | 125 | 125 | |
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1000 | 500 | 250 |
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500 | 250 | 250 | |
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1000 | 250 | 250 |
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250 | 125 | 125 | |
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>2000 | >2000 | 2000 |
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500 | 250 | 250 | |
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>2000 | >2000 | 1000 |
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500 | 250 | 125 | |
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62.5 | 62.5 | 62.5 |
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500 | 250 | 250 | |
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1000 | 500 | 500 |
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500 | 250 | 250 | |
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>2000 | >2000 | 1000 |
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500 | 250 | 250 | |
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1000 | 500 | 125 |
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250 | 250 | 250 | |
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500 | 250 | 250 |
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500 | 250 | 250 | |
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250 | 250 | 125 |
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500 | 250 | 250 | |
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500 | 250 | 250 | ||||
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>2000 | >2000 | 1000 |
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250 | 250 | 125 | |
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1000 | 125 | 62.5 |
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>2000 | 1000 | 500 | |
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1000 | 1000 | 2000 |
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1000 | 1000 | 1000 |
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500 | 250 | 62.5 | |
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1000 | 1000 | 2000 |
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250 | 250 | 62.5 | |
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2000 | 2000 | >2000 |
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500 | 250 | 62.5 | |
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1000 | 2000 | 2000 |
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2000 | 1000 | 1000 | |
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>2000 | 2000 | 1000 |
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2000 | 1000 | 1000 | |
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500 | 125 | NG |
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62.5 | NG | NG | |
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2000 | 1000 | NG |
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>2000 | >2000 | 2000 | ||||
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1000 | 500 | 250 |
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500 | 125 | 62.5 | ||||
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125 | 125 | 62.5 |
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500 | 500 | 250 | |
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1000 | 1000 | 500 |
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2000 | 250 | NG | |
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1000 | 500 | 500 |
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500 | 250 | 250 | |
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1000 | 1000 | 500 |
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250 | 250 | 250 | |
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>2000 | >2000 | 2000 |
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125 | 125 | 250 | |
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2000 | 1000 | 62.5 | |||||
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2000 | 2000 | 500 | |||||
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1000 | 1000 | NG | |||||
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2000 | 250 | NG | |||||
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500 | 250 | NG | |||||
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500 | 500 | 250 | |||||
NG : No growth recognized without yucca extract |
Table 8.
Antiyeast activity of yucca extract against 64 yeasts isolated from foods and effect of NaCl on antiyeast activity of yucca saponin fraction
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USFA* | TG** | |||||||
antiyeast activity | ↑a | ↑ | ↑ | ↓ | ↓ | ↓ | → | ↑ |
Table 9.
Effects of the cultural condition against antiyeast activity of yucca extract
*unsatulated fatty acid, **triglycerides, ***osmotic pressure
a:↑strengthen, ↓reduce, →no change
12. Utilization of the yucca extract as an anti-food deteriorating agents
Yucca extract is non-toxic and non-mutagenic. It is recognized as safe for human food use by U.S.FDA (listed in 21 CFR 172.510). The extract is tasteless and odourless, exerting no influence on the taste of foods. It is readily soluble in water and stable on heating. Based on the present study, commercial application of the extract for extending the shelf life of cooked foods and fermented seasonings is now under development [26].
Figure 4 shows the application of yucca extract to sponge cake. Addition of 0.2% of yucca extract to sponge cake had effective on the growth of fungi and yeasts stored in room for one week.
The application of yucca extract to strawberry jam was showed in Figure 5. The jam mixed 0.02% and 0.04% of yucca extract and stored in room for one week shows no change, whereas control jam was contaminated by fungi.

Figure 4.
Application of yucca extract to sponge cake

Figure 5.
Application of yucca extract to strawberry jam
13. Conclusion
The microbial safety of foods and cosmetics continues to be a major concern to consumers, regulatory agencies and food industries throughout the world. Although synthetic antimicrobials are approved in many countries, the recent trend has been for use of natural preservatives, which necessitates the exploration of alternative sources of safe, effective and acceptable natural preservatives. Many plant extracts possess antimicrobial activity against a range of bacteria, yeast and fungi, but the variations in quality and quantity of their bioactive constituents is major disadvantage to their industrial uses.
Based on the present study, mukurozi extract and yucca extract are considered to be effective for the preservation of foods and cosmetics. Both mukurozi and yucca plants have been consumed by humans for a long time. These plants also have wide application due to little pH or food component interaction.
Thus our works demonstrate that the saponin fraction from Sapindus pericarps and Mohave yucca stems can be recommended as alternative preservations for foods and cosmetics.
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