• 한국미생물·생명공학회지
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• 제24권4호
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• pp.430-438
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• 1996

The eleven strains, which could be used lactic and acetic acids as carbon sources, were isolated from kimchi and identified; the strains were facultative microorganisms which could be grown at low temperature (10$\circ$C) and around pH 3.2. As results of morphological, biochemical and physiological tests, 5 species of 3 genera were identified as Debaryomyces coudertii, Pichia edia, Pichia chambardii, Pichia haplophilia and Saccharomyces fermentati. Each strain was grown in basal media. In acidic resistance and acid utilization test, Saccharomyces sp. YK- 17 and Saccharomyces fermentati YK-19 were grown well in basal and YM media containing 0.3% lactic acid. And two strains were grown in basal and YM media containing O.3% lactic acid and 0.6% acetic acid. Since strain YK-19 was grown better at 10$\circ$C than that in 25$\circ$C, strain YK-19 was known to be a psychrophilic strain.

• 한국미생물·생명공학회지
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• 제24권4호
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• pp.512-518
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• 1996

Eleven strains had been previously isolated from kimchi and identified in our laboratory. The ability of each strain in aroma production was investigated by sensory evaluation. Among them Saccharomyc s sp. YK-17, Saccharomyces sp. YK-18, Saccharomyces sp. YH-3 and Saccharomyces fermentati YK-19 produced fruity flavour. Especially, Saccharomyces fermentati YK-19 produced apple and pineapple-like flavours. Pichia media YK-11, Saccharomyces sp. YK-20 and Pichia chambardii YH-4 produced wine-like flavour. Debaryomyces sp. YK-6, Debarymyces coudertii YK-10, Saccharomyces sp. YK-12 and Pichia haplophilia YH-5 produced alcoholic flavours. Using the good flavour producing strains as starters, various groups of kimchi were fermented, and the sensory characteristics of each group such as odor, taste and total acceptability were evaluated. The acidic odor, moldy odor and taste were reduced by adding starter, while the fresh sourness odor and taste similar to fruity fiavour were increased by starter. Comparing with the control group, these odor, taste and total acceptability were increased in the starter-added groups, such as Pichia edia YK-11, Saccharomyces sp. YK-17, and Saccharomyces,fermentati YK- 19. Saccharomyces fermentati YK-19 added kimchi group was higher siginificantly (P<0.05) than the others at the total acceptability. Volatile compounds of the culture broth of Saccharomyces fermentati YK-19 were analysed by gas chromatography, and 6 species of esters and 4 species of alcohols were identified. Among them, the ester substances which broth largely responsible for the apple-like flavour in the sensory evaluation, were found to be ethyl 2-methyl butvrate, ethyl pentanoate and ethyl acetate.

• Journal of Dairy Science and Biotechnology
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• 제39권3호
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• pp.87-93
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• 2021

This study analyzed and compared growth characteristics under large-scale fermentation at 35℃ of three microorganisms with the ability to reduce odor-producing substances in livestock. The three microorganisms (Bacillus subtilis FWC1, Bacillus amyloliquefaciens NAAS1, and Pichia farinosa NAAS2) evaluated in this study have been proven effective in reducing odor-inducing substances. Bacillus subtilis FWC1 exhibited the highest viable cell count when using 2% maltodextrin as carbon source, 0.05% soy-peptone as nitrogen source, and 0.3% yeast extract. The optimum media composition for B. amyloliquefaciens NAAS1 was 1.2% modified-starch with 0.8% yeast extract. The spore formation rate in the mass production of the Bacillus strains was over 90%, indicating that optimal growth medium compositions have been identified. In the case of P. farinosa NAAS2, our optimized growth medium [2% (w/v) glucose and 1% (w/v) yeast extract] improved biomass production.

• Food Science and Biotechnology
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• 제17권3호
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• pp.514-518
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• 2008

After overproduction of a recombinant $\beta$-galactosidase of Bifidobacterium infantis in Pichia pastoris, a synthesis of galacto-oligosaccharides (GOS) from 36% lactose using the enzyme (170.74 U/mg) was investigated. The transgalactosylation ratio reached up to 25.2% with 83.1% conversion of initial lactose and the maximum yield of GOS was 40.6%. The GOS syrup was composed of a 13.43% galacto-oligosaccharides, 5.06% lactose, and 8.76% monosaccharides. The prebiotic effect of GOS on the growth of bifidobacteria and lactobacilli strains was investigated in vitro. The maximum growth rate of Bifidobacterium breve and Lactobacillus acidophillus in GOS syrup (5%, v/v) media were 0.49 and 0.96/hr that are higher than those in 1%(w/v) galactose and 1%(w/v) lactose containing media. However, there was no significant difference between the specific growth rates of L. acidophillus in 1%(w/v) glucose and 5%(v/v) GOS syrup. Our data showed that GOS definitely promoted the growth of B. breve ATCC $15700^T$ and L. acidophilus ATCC 33323.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권1호
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• pp.43-51
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• 2002

The cloning and expression of $\beta-glucosidase$ II, encoded by the gene ${\beta}glu2$, from thermotolerant yeast Pichia etchellsii into Escherichia coli is described. Cloning of the 7.3 kb BamHI/SalI yeast insert containing ${\beta}glu2$ in pUC18, which allowed for reverse orientation of the insert, resulted in better enzyme expression. Transformation of this plasmid into E. coli JM109 resulted in accumulation of the enzyme in periplasmic space. At $50^{\circ}C$, the highest hydrolytic activity of 1686 IU/g protein was obtained on sophorose. Batch and fed-batch techniques were employed for enzyme production in a 14 L bioreactor. Exponential feeding rates were determined from mass balance equations and these were employed to control specific growth rate and in turn maximize cell growth and enzyme production. Media optimization coupled with this strategy resulted in increased enzyme units of 1.2 kU/L at a stabilized growth rate of $0.14\;h^{-l}$. Increased enzyme production in bioreactor was accompanied by formation of inclusion bodies.

• Journal of Microbiology and Biotechnology
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• 제20권5호
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• pp.946-949
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• 2010

The effects of two different sugars (glucose and xylose) on the expression levels and patterns of the xylose reductase (xyl1), xylitol dehydrogenase (xyl2), and xylulokinase (xyl3) genes were analyzed using Pichia stipitis. A significant increase in mRNA levels of xyl1 was observed after 6 h growth in culture conditions using xylose as a sole carbon source, but expressions of the three genes were not influenced by normal culture media with glucose. In addition, expressions of xyl2 and xyl3 were not observed during the entire culture period during which xylose was added. It also was found that the expression level of xyl1 increased as a function of the xylose concentration (40, 60, and 80 g/l) used in this study, indicating that xyl1 expression sensitively responded to xylose in the culture media. Although the induced level of xyl2 increased slightly after 48 h in the xylose-supplemented culture conditions, the expression of xyl2 was not observed in the xylitol-supplemented culture conditions. Finally, considering the expression of each gene in response to glucose or xylose, the absolute expression levels of the three genes indicate that xyl1 is induced primarily by exposure to xylose.

• KSBB Journal
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• 제25권4호
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• pp.351-356
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• 2010

리르노셀룰로오스로부터 생산된 글루코오스와 자일로 오스의 혼합당을 동시에 발효하여 에탄올 생산을 증가시키며, 또한 에탄올 생산에서의 세포고정화의 영향과 ICR (immobilized cell reactor)을 이용한 혼합당에서의 에탄올 연속생산을 수행하였다. 고정화 P. stipitis를 이용한 플라스크에서 에탄올을 생산에 대한 혼합당과 질소원의 영향으로부터 5% 혼합당 (글루코오스/자일로오스 = 3:1)과 1% 질소원이 최적으로 타나났으며, 이때 생산된 에탄올 농도는 약 19-20 g/L이었다. 고정화된 P. stipitis을 이용하여 반복적 유가식배양 (repeated fed-batch)으로 에탄올을 생산하였을 때는 모든 당 농도에서 글루코오스는 빠르게 소비되었지만, 혼합당의 농도가 높아질수록 자일로오스의 소비속도는 점차적으로 감소하였다. 즉 혼합당 농도가 증가하면서 더불어 당 소비속도는 감소하였다. 또한 ICR에서 1% 혼합당을 연속적으로 공급하면서 에탄올을 안정적으로 생산하여, 에탄올 농도는 5.6 g/L이었고 에탄올 생산 속도는 0.13 g/$L{\cdot}h$이었다.

• KSBB Journal
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• 제16권1호
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• pp.15-23
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• 2001

The development of expression systems for heterologous proteins has been greatly demanded not only for the study of the structure/function relationships of these proteins but also for their biotechnological and pharmaceutical applications. During the past decades, the methylotrophic yeast Hansenula polymorpha and Pichia pastoris have drawn attention as one of promising hosts for the production of a variety of heterologous proteins. The increasing popularity of H. polymorpha and P. pastoris as the host systems can be attributed to the several advantages over the traditional yeast Saccharomyces cerevisiae, such as the availability of very strong and tightly regulated promoters from the enzymes involved in the metabolism of methanol, a very high-cell density even on simple mineral media, and a high stability of expression plasmids. Furthermore, it has been observed that glycoproteins from these two yeasts are less hyperglycoylated compared to those from S. cerevisiae. Despite substantial similarities as methylotrophic yeasts, however, these two expression systems have some unique features distinguished from each other. In this paper we present a brief overview on the present status of the expression systems developed in methylotrophic yeast, mainly focusing on the similarities and differences between the H. polymorpha and P. pastoris systems.

• Applied Biological Chemistry
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• 제13권1호
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• pp.97-103
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• 1970

간장국제조과정중에 생육하는 효모들을 계수 및 분리 동정하고 TTC 정색에 의하여 유발한 결과는 다음과 같다. 1. 간장국 1g 중의 효모수는 3일국에 $97{\times}10^3$, 4일국에 $135{\times}10^3$, 5일국에 $179{\times}10^3$개로서 제국일수의 경과에 따라 효모수가 증가하였다. 2. 이들 효모의 TTC 정색에 었어서 White가 53.6${\sim}$71.9%, Red가 5.6${\sim}$11.1%, Red pink가 6.8${\sim}$1905 pink가 11.1${\sim}$22.6%로서 white 효모가 가장 많았고 Red 효모가 가장 적었다. 3. 30주를 임의 분리하여 동정한 결과 Saccharomyces rouxii 5주, Saccharomyces fermentati 2주, Saccharomyces rosei 1주, Hansenula suaveolens 1주, Hansenula anomala 6주, Pichia polymorpha 6주, Debaryomyces nicotianae 2주, Debaryomyces hansenii 2주, Torulopsis candida 2주, Torulopsis sake 2주, Candida pelliculosa 1주등 6속 11종으로 동정되었다. 4. 등정한 이들 효모를 TTC 정색에 따라 유별한 결과 Hansenula suaveolens, Hansenula anomala, Candida pelliculosa, Debaryomyces nicotianae, Pichia polymorpha 등은 국중에 생육하는 전체 효모의 약 60%를 차지하는 TTC white 효모구에 속하였으며 Saccharomyces rouxii와Saccharomyces rosei는 Red 로 Saccharomyces fermentati와 Debaryomyces hansenii는 Red pink로 Torulopsis candida와 Torulopsis sake는 pink로 나타났다. 5. 분리된 효모의 대부분이 무염 또는 식염10% 합유배지에서 잘 생육하나 15-18% 식염 합유배지에서는 생육이 거이 억제되었다.

• 한국미생물·생명공학회지
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• 제25권5호
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• pp.435-441
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• 1997

Forty seven strains of yeast were isolated from traditional Meju and were identified as Saccharomyces spp. (7 strains), Zygosaccharomyces spp. (7 strains), Kluyveromyces spp. and Hansenula spp. (each 5 strains), Rhodotorular spp. (8 strains), Candida spp. (12 strains), Pichia spp. and Debaryomyces spp. from results of their microbiological characteristics. The optimal medium for growth of all the yeasts was YM media and the optimal initial pH of the medium was 6.0. The optimum temperature for growth was 30$circ$ and among them, Sacch. exiguus OE-5, Sacch. cerevisiae OE-16, Sacch. kluyveri C-1 strains were thermotolerant yeasts which could grow at 40$circ$C.