• Journal of Microbiology and Biotechnology
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• v.3 no.1
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• pp.24-30
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• 1993

In order to develop a novel yeast which produces the charateristic aroma of soy sauce, a protoplast fusion between Zygosaccharomyces rouxii WFS4 and Torulopsis versatilis IAM 4993 was carried out. Auxotrophic mutants as selective markers were obtained from Zygosaccharomyces rouxii and Torulopsis versatilis by treatment of N-methyl-N -nitro-N-nitrosoguanidine. The conditions of the protoplast formation and the regeneration for fusion were examined. The protoplast fusion using polyethylene glycol 4000 led to the fusion frequency of $4~5{\times}10^{-7}\;cells/ml$. Among fusants, a fusant ST723-F31 presented the best results in terms of the aromaticity of fragrance, the growth pattern, the resistance against salt and the degree of growth according to pH. It makes easy to control the production and the balance of aroma components so that it gives a good flavor, shortens the fermentation period and, simplifies the preparation process when using a bioreactor into which fusant is immobilized.

• Korean Journal of Food Science and Technology
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• v.12 no.4
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• pp.313-323
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• 1980

The objective of this experiment was to improve the quality and shortening the aging time of Kochujang by adding mixed starter cultures of yeast strains. Kochujangs were mashed during the summer season with mixed starter cultures of Saccharomyces rouxii, Torulopsis versatilis and Torulopsis etchellsii. Enzyme activities and chemical composition of the Kochujang were determined during the period of aging and their organoleptic values were tested. The maximum activities of liquefying amylase and saccharogenic amylase in the Kochujang were obtained during 20 to 60 days and 20 to 30 days after mashing respectively. The acidic protease activity was reached maximum during 20 to 40 days. All enzyme activities were decreased markedly during the final stage of aging period. Among mixed starter cultures tested, mixed culture of T. versatilis and T. etchellsii shows the highest liquefying and saccharogenic amylase activities. Ethyl alcohol contents in 10 days after mashing were highest in the Kochujang with S. rouxii and T. versatilis, followed in order of S. rouxii and T. etchellsii mixture, T. versatilis and T. etchellsii mixture and control without addition of yeast. But the contents in all sample became similar after 20 days with the level of 2.3 to 2.8% and then decreased gradually. The level of reducing sugar contents was markedly increased during the first 10 days, especially in the batches of T. versatilis and T. etchellsii mixture and control. However, the concentration became similar in all samples after 40 days. The contents of amino nitrogen were increased markedly during the first 10 days then slowly up to 90 days. The rate was high in the Kochujang with T. versatilis and T. etchellsii when compared with others. The organoleptic values of all Kochujang made with addition of yeast starter cultures were superior to control, especially in flavor, taste and color. The Kochujang with T. versatilis and T. etchellsii marked the highest value. The data obtained from this experiment suggests that the quality of Kochujang could be improved by using starter culture of suitable yeast strains according to product characteristics and aging time.

• Journal of Food Hygiene and Safety
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• v.2 no.1
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• pp.15-20
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• 1987

A general screening test for the expression of antioxidative activity was performed on over 36 cultures belong to yeast isolated from soy sauce, Makkuli, and molasses. Antioxidative activities of yeasts were examined by measuring oxidation such as peroxide value and thiobarbituric acid value in fish oil. Of these cultures, Saccharomyces cerevisiae IFO 2114 were found to have strong antioxidative activity. Saccharomyces rouxii and Torulopsis etchellsii isolated from soy sauce showed the strongest antioxidative activity among yeasts. Pichia ohmerii isolated from Makkuli showed the strongest antioxidative activity and Candida versatilis isolated from molasses showed also relative strong antioxidative activity.

• Applied Biological Chemistry
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• v.22 no.2
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• pp.65-90
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• 1979

This study was conducted to establish the brewing method which would be useful for the production of Kochuzang. Kojis, which were made from various materials and microorganisms under a covered condition, were investigated and compared. Yeasts (Saccharomyces rouxii and Torulopsis versatilis) were added to Kochuzang, and the enzyme activity, microflora, chemical composition, nitrogen content, alcohol content and free sugars of Kochuzang were investigated and analysed. The results obtained are as follows: 1. Koji making (1) Glutinous rice-soybean group was superior to glutinous rice group in the saccharogenic and liquefying amylase activities of three day-Koji. (2) Protease activity (acid, neutral and alkaline) of glutinous rice-soybean Koji, which was inoculated with Aspergillus oryzae A, was increased till the 5th day, while other groups showed maximum activity after the 3rd day. (3) The maximum cellulose activity of Aspergillus oryzae B-Koji and A-Koji was observed after the 2nd day and the 3rd day, respectively. High cellulose activity of Aspergillus oryzae B-Koji and A-Koji was respectively shown in glutinous rice group and glutinous rice-soybean group at maximum. (4) Compared with glutinous rice Koji, glutinous rice-soybean Koji gave larger number of yeast and aerobic bacteria. 2. Kochuzang Fermentation (1) Each Kochuzang group shoved different liquefying and saccharogenic amylase activities. The highest activities were generally shown in 10 to 40 days after mashing and remarkably reduced in the last stage of aging. (2) Protease activities of each group were strong in order of acid, neutral and alkaline protease. Especially acid protease showed highest activity at the 40th to 50th day Kochuzang. (3) Each group showed maximum cellulase activity in the 40th and 50th day-Kochuzang and then decreased. (4) Osmophilic yeast of yeast-added Kochuzang after one-month aging was distinctively outnumbered compared with non-yeast-added Kochuzang, but two groups were similar after two months. (5) Yeast-added group and non-added group gave almost the same number of halophilic lactic acid bacteria in Kochuzang, but the non-added group gave slightly larger number of aerobic bacteria than the yeast-added group. (6) Amino nitrogen contents in all test group were increased rapidly till the 60th day of Kochuzang aged. After that the contents were increased slowly. (7) Ethyl alcohol contents of 20day-fermented Kochuzang were high in order of Saccharomyces rouxii-added group, Torulopsis versatilis-added group, Saccharomyces rouxii and Torulopsis versatilis mixed group and non-yeast-added group. But all test group showed about 2% in ethyl alcohol content after 40days of aging. (8) Alcohol content in the 7 month-aged Kochuzang of all test groups was high in order of ethyl alcohol, n-butyl alcohol, n-propyl alcohol and iso-propyl alcohol. Torulopsis versatilis-added group had the highest value of ethyl alcohol, n-propyl alcohol and n-butyl alcohol. (9) Reducing sugar in Kochuzang was increased after 20 days of aging compared with the 10days-ferment. The reducing sugar content in Saccharomyces rouxii-added group was distinctively small compared with that of other groups, decreasing after 30days of aging. (10) Rhamnose, fructose, glucose and maltose were isolated from the 10 day fermented Kochuzang. Raffinose was also found after 300 days-aged group, and fructose content was high in the 300days-aged Kochuzang. However, glucose content was smaller than that of 10days-fermented Kochuzang. (11) For the organoleptic tests of Kochuzang, taste, flavour and color of yeast-added group were superior to the non yeast-added group. Especially the complex yeast group among the yeast added groups were the best of all. Yeast-added group after 300 days of aging took higher paint in flavour test than that of non-added group. Therefore, brewing method like complex yeast added group seems to be advantageous for short time brewing Kochuzang.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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• pp.512-519
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• 1997

RNA accumulating strain of Torulopsis versatilis MT-1 was cultured in molasses medium for higher contents of RNA in cell. Yeast cells were harvested at logarithmic phase on synchronous culture. Yield of cells on dry base to input sugar was 59.5%. Crude protein content was 55.1% in cell. RNA content was 13.9%. Some problems found in the process for the preparation of yeast extracts were improved by the addition of culture broth of Streptomyces faecalis MSF which secrete RNase (5' nuclease and 5' adenylic acid deaminase). When the culture broth of S. faecalis MSF was added in autolysis process 46% of RNA in cell was converted to I and G(5' inosinic acid and 5' guanylic acid) in extract. By addition of 3-7% culture broth of S.faecalis MSF in autolysis or enzymolysis process at the start or early stage, RNA in extract was converted easily to I and G and protein in cells was easily extracted and hydrolyzed to amino acid. Taste of those yeast extracts was delicious. The yeasty smell in yeast extracts was removed. And cell debris was easily removed from extract.

• Journal of Nutrition and Health
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• v.13 no.1
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• pp.43-48
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• 1980

In order to identify the taste components of Kochuzang (red pepper paste), glutinous rice Kochuzang were prepared with addition of Saccharomyces rouxii and Torulopsis versatilis respectively. The nitrogen compounds were determined during the fermentation period at intervals and the amino acid components of 210 days aged glutinous rice Kochuzang were measured. Results are as follows; 1) The increment of amino-N and soluble-N of Kochuzang were increased remarkably during fermentation period untill 60 day but after that period, the increment was slowly changed. 2) Ammonia-N of Kochuzang was increased slowly during aging period but after that time, the amount was decreased slightly. 3) Amino-N ratio and Nitrogen solubility of 300 days aged Kochuzang were 23.71-25.38% and 54.12-56.19% respectively. 4) 17 Kinds of amino acid were identified at 210 days aged Kochuzang those were Lysine, Histidine, Arginine, Aspartic acid, Threonine, Serine, Glutamic acid, Proline, Glycine, Alanine, Valine, Methionine, Isoleucine, Leucine, Phenylalanine, Tryptophane and Tyrosine. 5) Total amino acids were 3.5-4.1% which consists of Glutamic acid, 0.95-1.05%, Aspartic acid, 0.63-0.65% and Serine, Proline, Alanine, Valine, Leucine were 0.22-0.28% respectively but Methionine, Histidine and Arginine were below 0.1%, and Tyrosine, Tryptophane and Phenylalanine existed in Kochuzang in small quantities.