• Journal of Microbiology and Biotechnology
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• 제3권2호
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• pp.91-94
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• 1993

A 3-isopropylmalate dehydrogenase (3-IPMD) gene was cloned from Kluyveromyces fragilis. pJK104 could complement Escherichia coli leuB and Saccharomyces cerevisiae leu2 auxotrophs. The coding region was subcloned and the nucleotide sequence was determined. A 1.8 Kb EcoRI/SphI fragment of pJK104 subcloned in pUC18 could still complement the leuB mutation. An open reading frame of 1164 bp that corresponds to a polypeptide of 387 amino acids was found in the cloned fragment. The homology between the 3-isopropylmalate dehydrogenase of S. cerevisiae and that of K. fragilis was 68.13％ in nucleotides.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.131-137
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• 1995

The alkaline phosphatase (K-ALPase) gene of Kluyveromyces fragilis has been cloned (1) and determined its base sequences (2) previously in our laboratory. When the K-ALPase gene was expressed in Escherichia coli and Saccharomyces cerevisiae, it showed a constitutive activity in E. coli, and a derepressed activity in S. cerevisiae in phosphate-limited medium. Northem hybridization experiment was performed to elucidate the transcription level of the K-ALPase gene. Northern experiment showed that transcription level of K-ALPase gene in S. cerevisiae was higher in phosphate depletion, but it was higher in high phosphate medium than in phosphate limited medium in K. fragilis. The transcription initiation site of the K-ALPase gene was determined by primer extension analysis. It matched nucleotide position - 169 in relation to the putative trnslational start site.

• 한국미생물·생명공학회지
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• 제25권3호
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• pp.277-285
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• 1997

Kluyveromyces marxianus var. marxianus isolated as an inulin-assimilating microorganism produces inulin fructotransferase (inulaseII) which catalyses the conversion of inulin into di-D-fructofuranose 1, 2' : 2, 3' dianhydrde (DFAIII). The DFA produced by the organism was isolated by using active carbon column, and identified as DFAIII by high performance liguid chromatography. The culture medium giving maximum inulaseII production was found to consist of 1% sucrose and 0.75% yeast nitrogen base (YNB). The inulasell production was induced by inulin or sucrose as a carbon source and increased by addition of YNB as a nitrogen source. Optimal initial pH of the culture medium, culture temperature and medium volume for the enzyme production were pH 4.7, 30$\circ$C and 140 ml, respectively. Under the optimal conditions described above, the enzyme activity in the culture supematant reached 4.2 units/ml after cultivation for 36 h. The DFAIII was accumulated at 13.25 mg/ml after 48 h of culture in the Jerusalem artichoke tuber medium.

• 한국균학회지
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• 제41권3호
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• pp.181-184
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• 2013

충남 금산군과 전남 완도군, 제주도 송악산 등지에서 서식하고 있는 야생화들로부터 Rhodosporidium fluviale 201-C-1과 Kluyveromyces thermotolerans 150-JE-2(1), Rodosporidium paludigenum과 Candida sp. 80-J-3등의 국내 미기록 효모들을 분리, 선발하여 이들의 미생물학적 특성들을 조사하였다. Rhodosporidium fluviale 201-C-1은 내염성의 무포자 효모로 의균사를 형성하였고 urease 활성을 갖고 있었다. Kluyveromyces thermotolerans 150-JE-2(1)는 내당성과 내염성을 가진 무포자효모로서 $30^{\circ}C$, pH 4.0~6.0에서 잘 생육하였다. Candida sp. 80-J-3과 Rhodosporidium paludigenum 86-J-1 균주들은 모두 무성생식을 하는 무포자 효모들이었고 YPD 배지, Vitamin-free 배지와 5% NaCl을 함유한 YPD 배지 등에서 잘 생육하였다. 또한 Candida sp. 80-J-3 균주의 urease 활성은 음성이었으나 Rhodosporidium paludigenum 86-J-1 균주는 urease 활성이 양성이면서 pH 7.0~8.0에서 잘 생육하였다.

• Applied Biological Chemistry
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• 제30권2호
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• pp.169-178
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• 1987

Kluyveromyces marxianus로 부터 inulase를 생산하고 정제하며 intra 및 extracellular inulase의 성질을 조사하였다. 본 균주는 stationary phase인 24시간째 intra 및 extracelullar enzyme의 생산이 최고에 달했으며 유기 질소원으로 YNB를 사용하고 배양 중 pH를 조절해 줌으로써 효소 생산을 향상시킬 수 있었다. 조효소는 DEAE-cellulose에 의해 intra 및 extracellular inulase 모두 2개의 fraction으로 분리되었고 각 fraction의 전기영동 양상은 비슷하여 주 band를 비롯 모두 3개의 glycoprotein band가 관찰되었으며 이중 주 band만 inulase 및 invertase activity를 보유하고 있었다. 정제 효소의 inulase 및 invertase의 최적 pH는 각각 5.0과 4.5였고 intra가 extracellular enzyme 에 비해 다소 넓은 범위의 pH에서 높은 활성을 나타내었다. 모든 fraction의 최적 온도는 inulase가 $40^{\circ}C$, invertase가 $50^{\circ}C$였으며 intracellular enzyme이 더 넓은 범위의 온도에서 안정하였고 열에 대한 안정성도 intracellular inulase가 extracellular inulase보다 높게 나타났다. Km value는 intra가 $16{\sim}19mM$, extracellular inulase가 $9{\sim}11mM$로써 extracellular inulase가 inulin에 대한 친화력이 더 높았으나 모두 exo-type의 inulase로 판명되었다.

• Applied Biological Chemistry
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• 제38권1호
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• pp.20-25
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• 1995

돼지감자 분말을 원료로 에탄올 생산성을 향상시키기 위한 전실험으로 Kluyveromyces marxianus FO43을 고정화시켜 sucrose 배지에 고정화 조건과 발효조건을 조사한 결과 alginate 농도는 2%, bead 직경 2 mm, 고정화 균주의 배양기간 24시간, particle input ratio of 30 : 100, 기질농도 10%(w/v), pH와 온도는 각각 5.5, $40^{\circ}C$가 최적으로 나타났으며 pH $4.5{\sim}6.5$, 온도 $30{\sim}45^{\circ}C$에서 높은 농도의 에탄올을 생성하여 고정화하지 않은 효모보다 넓은 pH와 온도 안정성을 보였다. 최적조건에서 회분발효를 실시한 결과 에탄올 농도는 46.4 g/L, 에탄올 생산성은 1.93 g/L.h이었고 최적조건에서 배양된 bead의 단면을 위상차 현미경으로 관찰해 본 결과 균주세포가 bead의 표면에 밀집되어 있었다.

• 한국미생물·생명공학회지
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• 제45권1호
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• pp.57-62
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• 2017

최근 모균주인 K. marxianus ATCC 36907 보다 xylose reductase의 활성이 증대된 돌연변이 K. marxianus 36907-FMEL1 균주를 개발하였다. 본 연구에서는 효율적인 xylitol의 생산을 위해 발효기를 이용하여 교반속도에 따른 효과를 확인하였다. $30^{\circ}C$ 조건에서 발효를 진행한 결과 K. marxianus 36907-FMEL1 균주는 400 rpm의 교반속도에서 가장 높은 수율 (0.57 g/g)과 생산성($0.64g{\cdot}l^{-1}{\cdot}h^{-1}$)을 확인할 수 있었다. 흥미롭게도 온도 조건을 $40^{\circ}C$로 증가하여 발효를 진행한 결과 agitation 조건 최적화를 통해 수율과 생산성이 각각 21% (0.64 g/g)와 58% ($0.90g{\cdot}l^{-1}{\cdot}h^{-1}$) 증가하였다.

• Food Science and Biotechnology
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• 제16권1호
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• pp.133-137
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• 2007

The Strain TFM-7, Which has an antitumor effect, was isolated from Kefir and identified based on analysis using the API 50 CHL kit and 265 rDNA sequencing. Strain TFM-7 was confirmed to belong to the genus Kluyveromyces. Analysis of the 265 rDNA nucleotide sequences found strain TFM-7 to be related to Kluyveromyces marxianus. NRRL Y-828IT. K. marxianus. TFM-7 was cultured with potato dektrose broth medium at $27^{\circ}C$ for 72 hr, and its inhibition effects on the proliferation of seven tumor cell lines and a normal cell line were assessed using the MTT assay. The antitumor effects and growth characteristics of K. marxianus TFM-7 were investigated during a culture period of 7 days. By the $3^{rd}\;day$, K. marxianus TFM-7 showed a dry cell weight 2.39 g/L, a pH of 4.39, an ethanol content of 0.89%, and an inhibition effect on the proliferation of seven tumor cell lines above 50%, except for A-549 tumor cell line. K. marxianus TFM-7 was the most effective at inhibiting the growth of Hep-2 cell line among all tumor cell lines tested. Growth inhibition of a normal cell line, NIH/3T3, was less than 35%, suggesting a decreased level of cytotoxicity toward normal cells. These results indicate that K. marxianus TFM-7 may have used as a yeast strain with antitumor activity.

• Journal of Microbiology and Biotechnology
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• 제3권1호
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• pp.6-11
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• 1993

An autonomously replicating sequence (Kf-ARS1) of Kluyveromyces fragilis was cloned from the genomic library which was constructed using pHN134 as a cloning vector to make a new host-vector system for the production of heterologous protein from K. fragilis as a host. The cloning vector pHN134 was composed of $Km^r, Ap^r$ and multiple cloning site in LacZ . A clone carrying Kf-ARS1 was isolated and the recombinant plasmid was designated as pIKD102. The cloned fragment was 2.3 kb (EcoRI/EcoRI) in length. Subcloning experiment showed that the region for ARS activity was 1.5 kb (SalI/EcoRI) fragment. It was shown that the Kf-ARS1 was active in Saccharomyces cerevisiae and Kluyveromyces fragilis.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1245-1253
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• 2014

One-step sulfuric acid saccharification of the red alga Pterocladiella capillacea was optimized, and various detoxification methods (neutralization, overliming, and electrodialysis) of the acid hydrolysate were evaluated for fermentation with the thermotolerant yeast Kluyveromyces marxianus. A proximate composition analysis indicated that P. capillacea was rich in carbohydrates. A significant galactose recovery of $81.1{\pm}5%$ was also achieved under the conditions of a 12% (w/v) biomass load, 5% (v/v) sulfuric acid, $121^{\circ}C$, and hydrolysis for 30 min. Among the various detoxification methods, electrodialysis was identified as the most suitable for fermentable sugar recovery and organic acid removal (100% reduction of formic and levulinic acids), even though it failed to reduce the amount of the inhibitor 5-HMF. As a result, K. marxianus fermentation with the electrodialyzed acid hydrolysate of P. capillacea resulted in the best ethanol levels and fermentation efficiency.