• Korean Journal of Agricultural Science
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• v.14 no.1
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• pp.156-163
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• 1987

In these experiment, the conditions of entrapping immobilization of Zygosaccharomyces rouxii that participate in the soysauce brewing are investigated. And carried out the fermentation and aging test by immobilized Zygosaccharomyces rouxii with the hydrolyzed solution prepared from soysauce koji. The results obtained were as follows: 1. Immobilizing conditions of Zygosaccharomyces rouxii. 1) When the concentration of Na-alginate solution is 2.0-2.5%, the bead formation was very good. And the concentration of Na-alginate solution not influenced on the fermentation activity of immobilized Zygosaccharomyces rouxii. 2) Effect of ratio of the precultured Zygosaccharomyces rouxii solution and Na-alginate solution on the fermentation activity of immobilized Zygosaccharomyces rouxii was not highly recognized. But if the ratio of precultured Zygosaccharomyces rouxii solution increased, the fermentation activity of immobilized Zygosaccharomyces rouxii was slightly high. 3) The fermentation activity of immobilized Zygosaccharomyces rouxii that grew over 36hrs was higher than that grew below 24hrs. 4) Increasing the ratio of immobilized Zygosaccharomyces rouxii gel to the fermentative medium, the fermentation activity of Zygosaccharomyces rouxii was higher. 2. The fermentation test by immobilized Zygosaccharomyces rouxii with the hydrolyzed solution of soysauce koji. 1) When fermented for about 96 hrs, the alcoholic fermentation almost stopped and alcohol concentration into the hydrolyzed solution of soysauce koji was 2.6%(v/v) approximately.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.136-141
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• 1988

Susceptibility of a thermophilic strain (D-71) of Saccharomyces cerevisiae and an osmotolerant strain (SR-S) of Zygosaccharomyces rouxii to Zymolyase-20T were studied in various renditions. Content of glucan and mannan in cell wall of Saocharomyces cerevisiae D-71 were 14.5% and 14.8%, and Zygosaccharomyces rouxii SR-S were 24.0% and 19.0%, respectively. Susceptibility of Saccharomyces cerevisiae D-71 cultured in Wickerham synthetic medium containing 0.5% of methionine and 0.1% of glucose to Zymolyase-20T was 66%, and $K_2$HPO$_4$ and aminobenzoic acid were greatly effective to susceptibility. Susceptibility of Zygosaccharomyces rouxii SR-S cultured in Wickerhnin synthetic medium containing 0.5% of peptone, 0.15% of methionine and 0.l% of glucose to Zymolyase-20T was 80%, and KI and pyridoxine were greatly effective to susceptibility. Susceptibility of Saccharomyces cerevisiae D-71 stationary cultured in YMPG medium at $25^{\circ}C$ for 12 hours was 16o1e and Zygosaccharomyces rouxii SR-S stationary cultured in YMPG medium at $25^{\circ}C$ for 30 hours was 82%.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.1
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• pp.101-107
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• 1993

Zygosaccharomyces rouxii is a salt-tolerant yeast which plays an important role during the ripening stage of soy sauce fermentation. Z. rouxii used in the experiment could grow in YPD (1 % yeast extract, 2% peptone and 2% glucose, pH5.0) medium with 18% (w/v) NaCl, whereas Saccharomyces cerevisiae could only grow in YPD medium with less than 8% NaCl. In the presence of 15% NaCl, Z. rouxii accumulates a large amount of glycerol as a compatible solute within the cells in the exponential phase. It is a characteristic of salt-tolerant yeasts. From the chemical analyses on membrane lipid fluidity, the membrane structure of the cells grown in 15% NaCl was suggested to become more rigid and its fluidity was decreased to keep glycerol within the cells in response to surrounding medium with high concentrations of salt.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.293-296
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• 1988

Stability of spheroplasts prepared from Saccharomyces cerevisiae D-71, a thermophilic strain and Zygosaccharomyces rouxii SR-S, an osmotolerant strain were studied. Stability of spheroplast from Saccharomyces cerevisiae D-71 was highest in 0.8M KCI and 1.0M sorbitol ; that from Zygosaccharomyces rouxii SR-S was highest in 0.4M KCI and mannitol and that from both strains was less than 10% for sonic oscillation at 20Kc for 60 sec. In centrifugation at 10000 x g for 10 min., stability of spheroplast from Saccharomyces cerevisiae D-71 was 93% and that from Zygosaccharomyces rouxii SR-S was 84%. Breakage of spheroplast from Saccharomyces cerevisiae D-71 was 99% and that from Zygosaccharomyces rouxii SR-S was 55% for UV irradiation with 15W UV lamp at a distance of 20 cm for 60 min.

• Korean Journal of Food Science and Technology
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• v.20 no.2
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• pp.164-169
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• 1988

Protoplast susion between Zygosaccharmoyces rouxii M-12 and Saccharimyces cerevusuare M-43 were investigated for breeding of a new brewing yeat strain for soy sauce. Auxotrophic mutants of Zygosaccharomyces rouxii ZRM-83 ($Met^-,\;Thr^-$) and Saccharomyces cerevisiae SCM-46 ($Lys^-,\;Arg^-$) were selected by treatment of 3.0% ethylmethane sulfonate and nutritional complementary method. Protoplast of both strains were more effective by treatment of 0.05mg/ml zymolase 20T for 60min. Fusion effeciency was much higher by treatment of 30% PEG 6,000 for 30min and fusion frequencies were $10^{-4}{\sim}10^{-5}$. These fusants originated from two protoplasts had properties of big cell size and much DNA content.

• The Korean Journal of Mycology
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• v.27 no.3 s.90
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• pp.181-186
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• 1999

Osmotolerant yeast isolated from soy paste could grow on media with 2 M NaCl. This strain was identified as Zygosaccharomyces rouxii by biological characteristics, RFLP of ribosomal DNA and mating with compatible haploid strain. Growing rate of the Z. rouxii YDJ was slower than Saccharomyces cerevisiae. Z. rouxii YDJ accumulated trehalose, which is known as one of the osmolytic protectants, in cells cultured on media with salt. Enzyme activity of trehalose phosphate synthase related to trehalose biosynthesis of the YDJ was lower than those of S. cerevisiae. Trehalase activity related trehalose degradation was also lower in Z. rouxii YDJ than S. cerevisiae. However, as Z. rouxii accumulated trehalose by salt treatment, salt tolerancy of Z. rouxii was assumed to be related to trehalose in additon to glycerol.

• KSBB Journal
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• v.18 no.6
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• pp.455-460
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• 2003

This study was designed to find out the rapid fermentation of soy sauce from koji hydrolyzates using air bubble column reactor packed with coimmobilized mixed culture system. Continuous ripid production was performed by coimmobilized Z. rouxiii BH-90 and C. versatilis BH-91. Coimmobilized cells of Zygosaccharomyces rouxii BH-90 and Candida versatilis BH-91 mixture cells in the column reactor produced 2.8％ ethyl alcohol and 18mg/L 4-ethylguaiacol over 96 hours under the optimal conditions. Coimmobilized cells produced 2.30∼2.4% ethyl alcohol during 30 days, and decreased gradually from 40 days to 70 days. Also coimmobilized cells produced 4-ethylguaiacol at the constant rate of 16∼18mg/L and decreased gradually after 40 days. Final product of soy sauce contained 2.4% ethyl alcohol and 18mg/L 4-ethylguaiacol. However, amino acid compositions of soy sauce were consisted of predominantly glutamic acid, leucin, arginine, aspartic acid, Iysine and valine, which were more than 50% of total amino acid.

• Microbiology and Biotechnology Letters
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• v.22 no.2
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• pp.120-125
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• 1994

By using a synthetic medium simulated on the amino acid composition of soybeam, the effect of salt concentrations on the production of volatile organic acid by the strains of Zygosaccharomyces rouxii So-3101, a soy sauce yeast, was studied at the concentrations of 12.5, 18.0, 22.0 and 28.5% NaCl. The growth, consumption of glucose, and production of alcohol, total acid and volatile organic acid, showed the highest values at a concentration of 12.5% NaCl, and those values were decreased with an increase in the salt concentration. The ratio of volatile organic acid to total organic acid was remained at approximately the same level within the range of salt concentrations between 12.5~22.0%, whereas the ratio was decreased at a salt concentration of 28.5%. After incubation for 16 days, 8 volatile organic acids, i.e. acetic, propionic, n-butyric, isobutyric, isovaleric, isocaproic, n-caproic, and heptanoic acids, were detected by gas chromatography. Among the volatile organic acids, acetic acid was produced in the appreciable amiunt and its ratio to the other volatile acids was increased with an increase in the salt concentration.A small amount of isocaproic, propionic, isobutyric and isovaleric acids were produced, and n-caproic, n-butyric and heptanoic acids were detected only at the lower salt concentration.

• Korean Journal of Food Science and Technology
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• v.28 no.2
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• pp.253-259
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• 1996

To investigate the yeast florae in the traditional and commercial Kochujang, computer identification systems, Vitek, API kit and conventional identification methods were used. Yeast florae of each process were compared and their typical physiological characteristics were also tested. Various process intervals yielded 330 colonies, which resulted in 11 species 184 strains classified. They were identified into Candida glabrata C. guilliermondii. C. humicola. C. rugosa, C. zeylanoides, Cryptococcus uniguttulatus, Pichia farinosa, Rhodotorula glutinis, Saccharomyces cerevisiae and Zygosaccharomyces rouxii. The strains of Candida, Pichia, Saccharomyces and Zygosaccharomyces were existing in both processes. In case of commercial process, the maximum distribution of Z. rouxii and S. cerevisiae were 33% at 15 day fermentation and 13% at 21 day, respectively. The distribution of Candida spp. was gradually decreased throughtout the fermentation period from 40% to 10%. In the traditional process, the maximum distribution of Z. rouxii and S. cerevisiae were 53% after 3 months and 26% after 7 months, respectively, S. cerevisiae and Z. rouxii showed distintive growth pattern at the high concentration of glucose and sodium chloride and played important roles in both processes of fermentation. Physiological tests revealed that only two major yeasts. S. cerevisiae and Z. rouxii, showed vigorous carbon dioxide formation under the tested conditions.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.366-372
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• 1993

The aim of this study was to find out the rapid fermentation of soy sauce from koji hydrolyzate using column type reactor packed with immobilized yeast cells. Each immobilized cell of Zygosaccharomyces rouxii BH-90 and Candida versatilis BH-91 in the packed column type reactor produced 2.8% ethyl alcohol and 18mg/l 4-ethylguaiacol over 96 hours under the optimal condition. Continuous fermentation was performed by immobilized Z. rouxii BH-90 packed in column type reactor. Immobilized Z. rouxii BH-90 produced 2.30~2.85% ethyl alcohol during 30 days, and decreased gradually from 40 days to 80 days. Also C. versatilis BH-91 produced 4-ethylguaiacol at the constant rate of 16~18mg/l and decreased gradually after 40 days. Final product of soy sauce contained 2.8% ethyl alcohol and 18mg/l 4-ethylguaiacol. However, amino acid compositions of final products were consisted of predominantly glutamic acid, leucine, arginine, aspartic acid, lysine and valine, which were more than 50% of total amino acid.