• Title/Summary/Keyword: yeast selection

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Optimization of Fermentation Conditions for Production of Recombinant Human Interleukin-2 in Escherichia coli (대장균에서의 재조합 인체 인터루킨-2 생산을 위한 발효조건 최적화)

  • Lee, In-Young;Kim, Myung-Kuk;Na, Doe-Sun;Hahm, Kyung-Soo;Moon H. Han;Lee, Sun-Bok
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.327-333
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    • 1988
  • For optimal production of recombinant human interleukin-2 (IL-2) in E. coli the effect of fermentation conditions on cell growth, IL-2 production, and stability of recombinant cells were investigated. Among the complex nutrients tested in this work, yeast extract, peptone and corn steep liquor were found to be effective for recombinant cell growth. The recombinant cells were maintained stably under repression condition (3$0^{\circ}C$), but the stability of recombinant cells were drastically reduced upon induction of IL-2 expression (42$^{\circ}C$) even under the selection pressure. Addition of antibiotics to the culture medium resulted in the cell growth inhibition without significant improvement in recombinant stability. When the expression of IL-2 gene was induced at different growth phases, highest IL-2 production was achieved by the induction of IL-2 at the middle-exponential growth phase. It was found that the production of IL-2 significantly inhibited the cell growth and the ex-pression of other genes in the plasmid.

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Yeast Selection and Comparison of Sterilization Method for Making Strawberry Wine and Changes of Physicochemical Characteristics during Its Fermentation (딸기주 발효를 위한 효모 선발과 살균 방법의 비교 및 발효 중 이화학적 특성의 변화)

  • Jeong, Eun-Jeong;Kim, Yong-Suk;Jeong, Do-Youn;Shin, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.38 no.5
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    • pp.642-647
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    • 2006
  • Yeast selection and the sterilization methods of strawberry juice were optimized for making strawberry wine. In addition, changes in the physicochemical characteristics of the wine during its fermentation were estimated. Maehyang and Yukbo cultivars of strawberry were tested for wine making; they contained 9.8 and 9.3% soluble solids and 0.59 and 0.58% titratable acidities, respectively. Among six yeasts tested, the Wg-15 and Sc-51 strains were selected based on the alcohol yield in the strawberry wine. Alcohol and soluble solid contents following heat treatment ($85^{\circ}C$, 10 min) or $K_2S_2O_5$ (200 ppm) treatment for sterilization were 7.10-7.20% and 5.60-5.80%, respectively, and no differences were observed between the Wg-15 and the Sc-51 strains. However, the flavor of wine produced following heat treatment was slightly better than that following $K_2S_2O_5$ treatment. The greatest amounts of alcohol were produced after 2 days of fermentation at $26^{\circ}C$. The alcohol content in wines produced with 12, 14, and 16% sugar content in the initial stages were 5.1, 6.0-6.2, and 7.5-7.7%, and the soluble solid contents were 3.9-4.3, 4.1-4.3, and 5.0-5.3%, respectively; no significant differences were observed between the Wg-15 and the Sc-51 yeast strains. For making strawberry wine, we proposed that the sugar content of Maehyang or Yukbo cultivars be adjusted to 16% soluble solids in the initial stages with heat treated at $85^{\circ}C$ for 10 min and fermentation with the Wg-15 or Sc-51 yeast strains at $26^{\circ}C$ for 8 days.

Selection of Inhibitor-resistance Yeast and its Application to Bioethanol Production in the Hydrolysate of Rape Stem (유채대 가수분해물에서 inhibitor 저항성 효모선별과 이를 이용한 bioethanol 생산)

  • Yeon, Ji-Hyeon;Kim, Hye-Ji;Oh, Sung-Ho;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • KSBB Journal
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    • v.25 no.4
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    • pp.401-407
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    • 2010
  • We established a strategy for bioethanol production using the hydrolysate of rape stem, in which the inhibitor cocktail was added intentionally. The final goal of this study was to circumvent the detoxification process when the hydrolysate of lignocelluloisic biomass contained the toxic substances in high concentration. When six yeast strains were examined, Sacchromyces cerevisiae ATCC 96581 and Pichia stipitis CBS 7126 were relatively resistant to inhibitor cocktail. Then, using strains 96581 and 7126, we designed a process strategy for bioethanol production, assuming that the concentration of toxic substance in the hydrolysate of rape stem was remarkably high. When strains 96581 and 7126 were inoculated simultaneously, it was observed that strain 7126 produced bioethanol as well as strain 96581, although the concentration of inhibitor cocktail was 18.2% (v/v). Finally, throughout this co-cultivation of strains 96581 and 7126, bioethanol was produced about 6.0 (g/L), and bioethanol yield reached at 0.4 (g-bioethanol/g-reducing sugar) (78.4% of theoretical value).

Enhancement of Ethanol Tolerance of Lactose Assimilating Yeast Strain by Protoplast Fusion

  • Ryu, Yeon-Woo;Jang, Heang-Wook;Lee, Haing-Sook
    • Journal of Microbiology and Biotechnology
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    • v.1 no.3
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    • pp.151-156
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    • 1991
  • In order to construct a yeast strain having high ethanol tolerance together with good lactose fermentation ability, the protoplast fusion using Saccharomyces cerevisiae STV 89 and Kluyveromyces fragilis CBS 397 was carried out. Auxotrophic mutants of K. fragilis were obtained as a selection marker by treatment of ethylmethane sulfonate. The best mutant for protoplast fusion was selected based on the capabilities of ${\beta}-galactosidase$ production and lactose fermentation. The protoplast fusion using polyethylene glycol and calcium chloride solution led to the fusion frequence of $3{\times}10^{-6}$ and a number of fusants were obtained. Among these fusants, a fusant F-3-19 showed the best results in terms of ethanol tolerance, ${\beta}-galactosidase$ activity and lactose fermentation. The performance of lactose fermentation and ethanol tolerance by this fusant were better than those of K. fragilis. Study on the ethanol tolerance having relation to fatty acid composition and intracellular ethanol concentration revealed that the fusant F-3-19 had a higher unsaturated fatty acids content and accumulated less amount of intracellular ethanol compared with a parent of K. fragilis.

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Selection and Target-Site Mapping of Peptides Inhibiting HCV NS5B Polymerase Using Phage Display

  • Kim, Min-Soo;Park, Chan-Hee;Lee, Jong-Ho;Myung, Hee-Joon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.328-333
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    • 2008
  • A series of pep tides binding to the HCV NS5B polymerase was selected from phage display peptide libraries. A conserved motif of Ser-Arg-X-Arg/Leu was identified among the selected peptides, and Pep2 (Trp-Ser-Arg-Pro-Arg-Ser-Leu) was chosen for further characterization. The binding of Pep2 to HCV NS5B in vivo was shown by a yeast two-hybrid assay and by subcellular colocalization analysis using immunofluorescence confocal microscopy. The in vitro interaction was also confirmed by GST pulldown assay. The replication of the HCV 1b subgenomic replicon was efficiently inhibited by the presence of the peptide. By using a subtractive biopanning against Pep2, the binding site of the peptide was mapped at the pocket of Pro388 to Pro391 in the thumb subdomain of the polymerase. A yeast two-hybrid analysis using Pro388Ala and Pro391Ala mutants of NS5B confirmed the binding.

Studies on The Production of The Single Cell Protein from Alcohol Distillation Slops of Cane Molasses(Part I) -on the selection and identification of the yeast- (당밀(糖密) 주정증류폐액을 이용한 단세포(單細胞) 단백질(蛋白質)의 생산에 관한 연구 (제1보) - 폐액 자화균(資化菌)의 분리(分離) 및 동정(同定))

  • Kim, Chan Jo;Oh, Man Jin
    • Korean Journal of Agricultural Science
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    • v.2 no.2
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    • pp.463-468
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    • 1975
  • 1) 224 strains were isolated from the accumulated soil and sewage samples flowing the waste of alcohol distillation, and among of them 2 strains of yeast were selected on the basis of their superior growth in the medium containing alcohol waste by shaking culture. 2) Morphological and physiological characteristics of the selected strains were investigated, and strain-73, strain-124 were identified Candida ciferrii, Candida brumptii by the manual of LODDER, respectively.

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In vitro selection of lactic acid bacteria for probiotic use in pigs (양돈용 생균제 개발을 위한 유산균주 선발)

  • Byun, Jae-won;Kim, Gyung-tae;Bae, Hyoung-suk;Baek, Voung-jin;Lee, Wan-kyu
    • Korean Journal of Veterinary Research
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    • v.40 no.4
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    • pp.701-706
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    • 2000
  • This study was carried out to select the lactic acid bacteria(Lactobacillus, Streptococcus and Bifidobacterium) and yeast for probiotic use in pigs. Acid-tolerant 536 strains were isolated from the feces of 30 pigs. To select useful strains, the first screened strains were treated with strong acid solution(pH 2.5 to 3.0) for 3 hours and subsequentely treated with the anaerobic diluent solution containing 0.15% Oxgall for 3 hours. Among these strains, 151 strains showed strong tolerance to both acid and bile. Lactobacillus and Streptococcus tolerant to the acid and bile were treated with heat at $80^{\circ}C$ for 15 min, and at $70^{\circ}C$ for 5 min in Bifidobacterium and yeast. As a result of heat treatment, 38 strains were obtained as heat-tolerant strains. All of heat-tolerant strains were tested for antibiotic resistance against virginiamycin, sulfathiazole, aureomycin, neomycin, linsmycin, tiamulin and ASP250 which were used as feed additives for growth promotion in pigs. Finally, one strain each from Lactobacillus, Streptococcus, Bifidobacterium and yeast that showed resistance to acid, bile, heat and antibiotics was selected for probiotic use in pigs.

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IN VITRO STUDY ON THE ADHERENCE AND PENETRATION OF CANDIDA ALBICANS INTO DENTURE SOFT LINING MATERIALS (의치 연성이장재에 대한 Candida albicans의 부착과 침투연구)

  • Kim Min-Ju;Shin Sang-Wan;Lee Jeong-Yeol
    • The Journal of Korean Academy of Prosthodontics
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    • v.44 no.4
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    • pp.466-476
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    • 2006
  • Purpose : Colonization of denture soft lining materials by Candida albicans can result in clinical problem, and deterioration of the materials. This study aimed to compare the retention and penetration of C. albicans into four denture soft lining materials commonly used. Materials and methods : Four denture soft lining materials (Coe-comfort$^{(R)}$, Coe-soft$^{(R)}$, GC soft liner$^{(R)}$, and Tissue conditioner$^{(R)}$) discs were prepared to glass slide and dental stone. Adherence of yeast to surfaces was monitored after one hour incubation of standardized washed cell suspension with test disc surfaces. Adherent cells stained with acridine orange were counted fluorescence microscopy. Penetration of yeast into materials bonded with acrylic resin after 1, 2, 3,4, 5, 6 and 7 days incubation was observed through sections stained using acridine orange and estimated to quantitative analysis using radioisotope. Results : There was statistical significance in cell numbers between smooth and rough surfaces(p<0.05). Higher numbers of cells were observed on rough surfaces. There was statistical significance in adherent cell numbers into smooth and rough surfaces individually(p<0.05). According to the increase of incubation periods, the cells penetrated into denture soft lining materials were shown to increase. The differences among all kinds of soft liner were statistically significant(p<0.05),and the largest number of cells penetrated into soft liners was observed in the Coe-soft$^{(R)}$. Conclusion : Initial adherence and penetration of yeast into denture soft lining materials has been influenced by surface roughness and chemical composition of them. The selection of appropriate materials and their fabrication may promote clinical performance.

Effect of supplementation of yeast with bacteriocin and Lactobacillus culture on growth performance, cecal fermentation, microbiota composition, and blood characteristics in broiler chickens

  • Chen, C.Y.;Chen, S.W.;Wang, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.211-220
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    • 2017
  • Objective: The aim of the present study was to investigate the effect of yeast with bacteriocin and Lactobacillus cultures (mixture of Lactobacillus agilis BCRC 10436 and Lactobacillus reuteri BCRC 17476) supplements, alone or in combination, on broiler chicken performance. Methods: A total of 300, 1-d-old healthy broiler chickens were randomly divided into five treatment groups: i) basal diet (control), ii) basal diet+0.25% yeast (Saccharomyces cerevisiae) (YC), iii) basal diet+0.25% yeast with bacteriocin (BA), iv) basal diet+Lactobacillus cultures (LAB), and v) basal diet +0.25% yeast with bacteriocin+Lactobacillus cultures (BA+LAB). Growth performance, cecal microbiota, cecal fermentation products, and blood biochemistry parameters were determined when chickens were 21 and 35 d old. Results: The supplementation of YC, BA, and BA+LAB resulted in a significantly better feed conversion rate (FCR) than that of the control group during 1 to 21 d (p<0.05). The LAB supplementation had a significant effect on the presence of Lactobacillus in the ceca at 35 d. None of the supplements had an effect on relative numbers of L. agilis and L. reuter at 21 d, but the BA supplementation resulted in the decrease of both Lactobacillus strains at 35 d. The BA+LAB supplementation resulted in higher short chain fatty acid (SCFA) in the ceca, but LAB supplementation significantly decreased the SCFA at 35 d (p<0.05). All treatments tended to decrease ammonia concentration in the ceca at 21 d, especially in the LAB treatment group. The BA supplementation alone decreased the triacylglycerol (TG) concentration significantly at 21 d (p<0.05), but the synergistic effect of BA and LAB supplementation was required to reduce the TG concentration at 35 d. The YC supplementation tended to increase the plasma cholesterol at 21 d and 35 d. However, the BA supplementation significantly decreased the cholesterol and low density lipoprotein cholesterol level at 35 d. In conclusion, the BA+LAB supplementation was beneficial to body weight gain and FCR of broiler chickens. Conclusion: The effect of BA and LAB supplementation may be a result of the growth of lactic acid bacteria enhancement and physiological characterization of bacteriocin, and it suggests that the BA and LAB supplementation level or Lactobacillus strain selection should be integrated in future supplementation designs.

Profiling of Metabolites and Proteins from Eschscholtzia californica induced by Yeast Extract (Yeast Extract로 처리된 Eschscholtzia californica의 Metabolite와 Protein의 변화)

  • Cho Hwa-Young;Park Jeong-Jin;Yoon Sung-Yong;Part Jong Moon
    • KSBB Journal
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    • v.20 no.4
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    • pp.285-290
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    • 2005
  • Benzophenanthridine alkaloids - sanguinarine, chelirubine, macarpine, and chelerythrine are produced from Eschscholtzia californica (Californica Poppy, used as a sedative by Native Americans) and most of them are derived from dihydrosanguinarine. The properties of sanguinarine are the basis of its antimicrobial activity and its use in chemosurgery and skin cancer excision. For overproduction of sanguinarine from E. californica, yeast extract was used as elicitor and the elicited cell's metabolites were checked. Sanguinarine production was increased intracelluarly about 8 times in the cell and 5 times extracelluarly. We have peformed proteomic analysis of proteins sequentially extracted from E. califormica suspended cells which were cultured with elicitor, an increase of spot intensity was seen at 24 hours following elicitation. These proteins were separated by two-dimensional electrophoresis (2-DE). We found several spots that were expected to be related to benzophenanthridine alkaloids production by comparing the production profiles of metabolites such as sanguinarine. These results demonstrate the use of metabolite analysis as a tool for detecting target proteins related to metabolites production pathway.