• Microbiology and Biotechnology Letters
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• v.24 no.4
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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.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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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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• v.39 no.3
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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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• v.17 no.3
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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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• v.7 no.1
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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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• v.20 no.5
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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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• v.25 no.4
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• pp.351-356
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• 2010

To increase the production of ethanol by using sugar from lignocellulosic biomass, pentose and hexose have to be fermented simultaneously by yeast. The effects of mixed sugar and nitrogen on ethanol production by immobilized Pichia stipitis KCCM 12009 were investigated. When optimal mixed sugar and nitrogen concentration were 5% (Glucose/Xylose = 3:1) and 1%, respectively, ethanol concentration produced by immobilized P. stipitis was 19-20 g/L. In repeated fed-batch by immobilized P. stipitis, all glucose was consumed very quickly at 1-3% mixed sugar concentration. But, xylose consumption was decreased as the mixed sugar concentration increased. Also, ethanol (5.6 g/L) was stably produced and ethanol production rate was 0.13 g/$L{\cdot}h$ in immobilized cell reactor (ICR) with 1% mixed sugar (Glucose/Xylose = 3:1) as feeding media.

• KSBB Journal
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• v.16 no.1
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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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• v.13 no.1
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• pp.97-103
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• 1970

This experiment was carried out to investigate the yeasts for the brewing of soy sauce. The yeasts in the soy sauce koji were isolated and identified. And they were classified by coloring with the treatment of TTC(2, 3, 5 triphenyltetrazolium chloride) agar and counted. The result, obtained was as follows; a) The number of the yeasts in the Koji was increased in process of incubation time: about $97{\times}10^3$ per gram of Koji incubated for 3 days, $135{\times}10^3$ 4 days and $179{\times}10^3$ 5days. b) They were classified as 53.6-71.9 per cent of TTC while yeasts for the most, 5.6-11.1 per cent of red yeasts, 6.8-19.5 per cent of red pink yeasts and 11.1-22.6 per cent of pink yeasts. c) 30 strains of yeasts were isolated from the Koji optionally, and they were identified as 6 genera and 11 species: 5 strains of Saccharomyces rouxii, 2 strains of Saccharomyces fermentati, 1 strain of Saccharomyces rosei, 6 strains of Saccharomyces rosei, 6 strains of Hansenula anomala, 1 strain of Hansenula suaveolens, 6 strains of Pichia polymorpha, 2 strains of Debaryomyces hansenii, 2 strains of Debaryomyces nicotianae, 2 strains of Torulopsis candida 2 strains of Torulopsis sake and 1 strains of Torulopsis sake and 1 strain of Candida pelliculosa. d) The yeasts belonging to Hansenula suaveolens, Hansenula anomala Candida pelliculosa, Debaryomyces nicotianae and Pichia polymorplza were classified as TTC white yeasts, Saccharomyces rouxii and Sacharomyces rosei were red, Saccharomyces fermentati and Debaryomyces hancenii were red pink, and Torulopsis candida and Torulopsis sake were pink. e) Generally the growth of the yeasts isolated were rapid on the media containing none or 10 per cent of sodium chloride but almost restrained on the media containing 15-18 per cent of sodium chloride.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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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.