• Title/Summary/Keyword: Alcohol yeasts

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Enhanced and Targeted Expression of Fungal Phytase in Saccharomyces cerevisiae

  • LIM, YOUNG-YI;EUN-HA PARK;JI-HYE KIM;SEUNG-MOON PARK;HYO-SANG JANG;YOUN-JE PARK;SEWANG YOON;MOON-SIK YANG;DAE-HYUK KIM
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.915-921
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    • 2001
  • Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

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Fermentation Characteristics of Wine Yeast Strains for White Wine Making (백포도주 양조에 있어서 포도주 효모의 발효 특성)

  • Seoktae Jeong;Nami Goto;Park, Jonguck
    • Food Science and Preservation
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    • v.8 no.3
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    • pp.326-330
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    • 2001
  • The characteristics of used wine yeast strains were as follows, S6U showed low fermentation speed than those of other yeast strains, but this strain fermented completely later. The wine fermented by W-3 was very low contents of total acid, 0.75% and the ones fermented by UCD530 and AC- contained much extract, 3.26 mg/L and 3.22 mg/L respectively. The wine fermented by CEG and CS2 were predominant in yellowness, and EC1118 produced large amount of acetaldehyde, 49.9 mg/L than those of other strains. EC1118 and CY3079 displayed low methylene blue dyeing ratio, below 15%, meaning high alcohol tolerance yeast. UCD530 produced extremely high contents of glycerol, succinate and lactate compared with other strains. These properties revealed that UCD530 was a typical Saccharomyces bayanus species. The main organic acids produced by wine yeasts were pyruvate, lactate, succinate and acetate. The concentration of acetate in experimental wine could be divided into two parts, one group had concentration below 170 mg/L (UCD530, EC1118, AC-, CY3079, W-3), and the other had concentration up to 350 mg/L (S6U, CEG, CS2).

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Studies on the Grape Variety and the Selection of Yeast Strain for Wine-making in Korea (국내포도주(國內葡萄酒) 생산(生産)을 위(爲)한 포도(葡萄)의 품종(品種) 선택(選擇) 및 최적(最適) 효모(酵母) 균주(菌株)의 선발(選拔)에 관(關)한 연구(硏究))

  • Park, Yun-Hee
    • Applied Biological Chemistry
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    • v.18 no.4
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    • pp.219-227
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    • 1975
  • In order to investigate the possibility of wine-making with the grape variety cultivating in Korea, the basic analysises were carried out; analysis of the grape maturity during ripening period, the composition of grape juice, and the chemical components of wine from different varieties. Also the yeasts existing naturally on the grape were isolated and identified. For the selection of strain, the characteristics of 6 strains were studied. The results obtained were summarized as follows. 1. The reducing sugar content increased considerably during two weeks after the ordinary grape-gathering period. 2. The sugar content was highest in Muscat bailey A, which could be fermented naturally. The other varieties, Campbell Early, Steuben, and Alden needed chaptalisation for wine-making. 3. The permanaganate number and the methanol content of wine from all varieties were lower than french wine. 4. The sensory evaluation of wine showed that the pink wine was appreciated better than the red wine of same variety and the foxy taste of wine from hybrid grape influenced little to Korean. 5. The selected 6 strains were identified as Saccharomyces chevaliers, Saccharomyces capensis and Saccharomyces globosus. The strain No. 3 and No. 4 showed the most excellent characters for wine-making.

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Improvement of Antibacterial Activities of Bacteriocidal Yeasts Using the GPD Promoter (GPD 프로모터를 이용한 항균활성 효모의 활성증진)

  • Jang, Min-Kyung;Yu, Ki-Hwan;Kim, Nam-Young;Lee, Ok-Hee;Shin, Jae-Kyun;Jang, Hye-Ji;Lee, Seung-Woo;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.20 no.6
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    • pp.934-939
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    • 2010
  • We have previously reported recombinant productions of bacteriocins using yeast expression plasmid pAUR123, which contains the alcohol dehydrogenase (ADH) promoter, in Saccharomyces cerevisiae cells and their antibacterial activities. In order to improve the antibacterial activities of bacteriocidal yeast cells, a strong glyceraldehyde phosphate dehydrogenase (GPD) promoter gene of S. cerevisiae was amplified and inserted upstream into bacteriocin genes such as the OR-7, Subpeptin JM4-A or JM4-B gene in the corresponding recombinant yeast plasmid. Yeast cells transformed by the recombinant plasmid containing the GPD promoter represented higher antibacterial activities against both Gram positive B. subtilis and Gram negative E. coli cells compared to those transformed by the corresponding recombinant plasmid containing the ADH promoter. Thus, yeast cells harboring the recombinant plasmid containing the GPD promoter constructed in this study could be applied in the food preservative or animal feed industries.

Manufacturing of Korean Traditional Rice Wine, Makgeolli, Supplemented with Strawberry and Its Physicochemical and Microbial Properties during Fermentation (딸기를 첨가한 막걸리의 제조와 발효 과정 중 이화학적 및 미생물학적 특성)

  • Bae, Sang-Min;Han, Sang-Min;Choi, Jong-Myung;Lee, Jong-Soo;Kim, Ha-Kun
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.307-313
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    • 2016
  • To develop a functional strawberry Makgeolli, we produced Makgeolli using strawberry as an additive and then investigated its physicochemical properties. Among 7 different alcohol-fermenting yeasts, Saccharomyces cerevisiae JSK104 produced 17.4% ethanol on the 7th day of fermentation and was selected for use in the brewing of strawberry Makgeolli. Changes in physicochemical properties, numbers of yeast and lactic acid bacteria, and antihypertensive angiotensin-converting enzyme inhibitory activity were investigated during the fermentation of strawberry Makgeolli. The pH tended to decrease and the total acidity increased as the fermentation period elapsed. The ethanol content reached about 17% on the 7th day after fermentation, and the numbers of yeast and lactic acid bacteria reached a maximum on the 1st day of fermentation and then maintained a constant number. The antihypertensive angiotensin-converting enzyme activity reached a maximum after 5 days of fermentation and then was not significantly changed afterwards.

Characteristics of Ice Wine Fermentation of Freeze-Concentrated Campbell Early Grape Juice by S. cerevisiae S13 and D8 Isolated from Korean Grapes (포도로부터 분리한 S. cerevisiae S13 및 D8에 의한 캠벨 얼리 동결농축 과즙의 아이스와인 발효 특성)

  • Hwang, Sung-Woo;Hong, Young-Ah;Park, Heui-Dong
    • Food Science and Preservation
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    • v.18 no.5
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    • pp.811-816
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    • 2011
  • Cryoextraction (a freeze concentration using an instrument) can increase the sugar concentration in grape juice by reducing its water content, similar to the natural freezing of grapes for natural ice wine. In this study, fermentation of freeze-concentrated Campbell Early grape (Vitis labruscana) juice to 36 $^{\circ}Bx$ was carried out using Saccharomyces cerevisiae strains D8 and S13 isolated from Korean grapes. During the fermentation, strains S13 and D8 showed rapid sugar reduction and alcohol production compared with S. cerevisiae Fermivin$^{(R)}$ used as a control. After nine-day fermentation, the residual sugar contents were lower in W13(9.77%) and D8 wine(9.07) than that in Fermivin$^{(R)}$ wine(14.0%). Total acid content was high in the D8>S13>Fermivin$^{(R)}$ wine, in that order. The acetaldehyde content was highest in the D8 wine and lowest in the Fermivin$^{(R)}$ wine, among the three. The methanol content was slightly higher in the S13 and D8 wines than in the Fermivin$^{(R)}$ wine. In the sensory evaluation, the S13 wine exhibited the highest score in flavor and taste among the three wines. Both the two S13 and D8 wines exhibited higher scores than Fermivin$^{(R)}$ wine in overall preference.

Characterization of Protoplast Fusant between Killer Yeast and Alcohol-Fermenting Yeast (Killer 효모와 알콜 발효효모간의 원형질체 융합주의 특성)

  • 정기택;방광웅;김재근;송형익;정용진
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.55-64
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    • 1990
  • Cell volume and DNA contents of the fusants were similar to those of parents. Genetic stability of the fusants was increased when they were cultured on minimal medium (MM) rather than on complete medium (CM), and the fusants were stabilized by subculturing 7 generations each 7 day on MM agar. The finally selected fusants after being cultured for 6 months on CM were stable without segregation. The fusants could also form nuclein and ascospores, and show red and pink colors by the test of TTC colorization. Assimilability and fermentability of carbon sources of the fusants were similarto those of parents. The tolerance of KCl, NaCl, sodium propionate and cycloheximide showed the traits of one strain of parents. When the fusants were cultured for 72 hr and 60 hr in the medium containing 20% glucose and sucrose, respectively, the yield of ethanol for FWKS 260 was reached to 9.6 v/v% and 9.8v/v%, respectively. The sensitive strain Kyokai 7 was found to be killed entirely after cultivation of 48 hr by the killer toxin from the fusants. The recipient S 29 and Kyokai 7 were found to have neither L nor M dsRNA plasmid. However, K 52 and fusants had both L and M dsRNA plasmid of 4.7 kb and 2.5 kb, respectively. The curants treated by heat and cycloheximide did not contain M dsRNA plasmid, but had large amounts of L dsRNA plasmid of those of killer yeasts.

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Quality Changes in Kiwifruit Wines during Fermentation and Aging with Different Yeasts (효모 종류에 따른 참다래 와인의 발효 및 숙성 중 품질 변화)

  • Oh, Hyun-Jeong;Lim, Sang-Bin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.4
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    • pp.481-489
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    • 2017
  • Kiwifruit wine was prepared with the domestic new cultivars 'Jecy Sweet' and 'Jecy Gold', and quality characteristics were investigated during fermentation with Saccharomyces bayanus Lalvin and Saccharomyces cerevisiae Fermivin and aging for 120 days at $15^{\circ}C$. Total acidities were 0.94% and 1.22% for 'Jecy Sweet' and 'Jecy Gold' at the beginning, respectively, and increased gradually during fermentation; highest acidities were 1.49% and 1.26%, respectively, on the 6th day of fermentation regardless of yeast strain. Alcohol content increased greatly from the 4th day of fermentation and was highest (10.2%) in 'Jecy Sweet' fermented by S. bayanus Lalvin, followed by 9.2% in 'Jecy Sweet' fermented by S. cerevisiae Fermivin and 9.4% in 'Jecy Gold' fermented by Lalvin and Fermivin on the 12th day of fermentation. Soluble solid content was $24.8^{\circ}Brix$ at the beginning and decreased gradually during fermentation. The lowest soluble solid content was $9.7^{\circ}Brix$ on the 6th day of fermentation regardless of kiwifruit cultivar and yeast strain. Total phenols in 'Jecy Sweet' (1,127 mg/L) were 1.32-fold higher than those in 'Jecy Gold' (848 mg/L) and decreased greatly until the 6th day of fermentation, after which they increased slightly until the 12th day of fermentation. During aging, total phenols increased until the 30th day and were maintained for 120 days. Quality characteristics of kiwifruit wines were similar between the two yeast strains but were different between kiwifruit cultivars. Contents of ethanol and total phenolics were higher in 'Jecy Sweet' wine than in 'Jecy Sweet' wine.

Studies on the Yeasts for the Brewing of Soy sauce(Part 7) -Industrial utilization of Saccharomyces rouxii $T_9$ in the brewing of soy sauce- (간장발효에 관여하는 효모에 관한 연구 (제 7 보) -Saccharomyces rouxii $T_9$을 이 용한 간장의 발효시험-)

  • Lee, Taik-Soo;Lee, Suk-Kun;Chu, Young-Ha;Shin, Bo-Kyu
    • Applied Biological Chemistry
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    • v.14 no.2
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    • pp.121-129
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    • 1971
  • During the Brewing of Soy Sauce on an industrial scale, Saccharomyces rouxii $T_9$ was cultured and added to the soy sauce mash. The comparative experiments of soy sauce mash in non-addition and addition group of yeast were examined in this report. The yeast flora and chemical composition of Soy sauce mash through out the brewing were observed and the results obtained were as follows. (1) The number of Osmophilic yeast in one ml of soy sauce mash showed $185{\times}10^3$ 1 month after mashing and $750{\times}10^3$ 4 months after mashing in case of yeast group, while presented as $98{\times}10^3$ 1 month after mashing and $394{\times}10^3$ 4 months after mashing in case of non-yeast group. And the number of Osmphilic yeast in yeast group was twice of that in non-yeast group. (2) The number of ordinary yeast of TTC red group was shown as $2132{\times}10^3\;to\;3252 ×10^3$ 5 to 6 months after mashing in case of yeast group, while presented $752{\times}10^3\;to\;1251{\times}10^3$ in case of non-yeast group. And the yeast group was shown more than non-yeast group in ordinary, red pink and pink yeast number. (3) TTC red yeast were strongly appeared in both addition and non-addition group of yeast from 1 month after mashing to 6 months after mashing. (4) Though total nitrogen, pure extract, pH and buffer action contents of soy sauce showed similiar tendency in yeast and non-yeast group, alcohol and color density contents were highly appeared in yeast group and reducing sugar content was in non-yeast group respectively. (5) By the results of Organic function test of soy sauce mash, the difference of taste quality in yeast and non-yeast group were not evidently appeared, however, the appearance and flavour of the soy sauce were better in yeast group than in non-yeast group.

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Characterization of quality changes of whole super sweet corn (Zea mays saccharata Sturt.) during thermal sterilization for shelf-stable products (상온유통을 위한 가열살균 중의 통 초당옥수수의 품질변화 연구)

  • Lee, Yun Ju;Yoon, Won Byong
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.25-30
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    • 2019
  • This study investigated the quality changes in whole super sweet corn during thermal processing to extend its shelf-life. To minimize the reduction of unique texture of whole sweet corn after the sterilization, the alcohol sanitation applied and the cold point of a whole corn ear was determined using a computer simulation. The cold point was located between the corn kernel and the cob. The microorganisms on the surface of sweet corn were reduced by more than 1 log CFU/g after alcohol sanitation, then the whole corn was treated to satisfy the degree of sterilization ($F_{121.1}=4$). The quality of sterilized sweet corn was compared with the control that was treated with steaming. The quality changes of sterilized sweet corn during storage were monitored for 9 months at $25^{\circ}C$. The hardness was maintained within 30% of its initial value. The minimum of hardness was $464.50{\pm}103.35g$ and maximum of hardness was $514.50{\pm}81.83g$. The differences in the sugar content among the samples were found, but the sugar content of corn kernel remained within 30% of the control, ranging from $28.83{\pm}1.05$ to $34.36{\pm}0.42%$. The yellowness was higher than that of control by 5%. The maximum value of yellowness was $34.36{\pm}0.42$. The general bacteria and molds and yeasts in corn kernel stored at $25^{\circ}C$ were not detected after 9 months of storage at $25^{\circ}C$. Therefore, in this study, we have demonstrated that the thermal sterilized method extends the shelf-life of whole sweet corn with minimizing its quality changes over 6 months in room temperature.