• Title/Summary/Keyword: K.marxianus

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Simultaneous Saccharification of Inulin and Ethanol Fermentation by Recombinant Saccharomyces cerevisiae Secreting Inulinase

  • Kim, Youn-Hee;Nam, Soo-Wan;Chung, Bong-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.55-60
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    • 1998
  • Various Saccharomyces cerevisiae strains were transformed with a 2 ${\mu}$-based multicopy expression plasmid, pYIGP, carrying Kluyveromyces marxianus inulinase gene under the control of GAPDH promoter. Among then two strains, SEY2102 and 2805, showed high levels of cell growth and inulinase expression, and were selected to study their fermentation properties on inulin. Jerusalem artichoke inulin was more effective for cell growth (10∼11 g-dry wt./L at 48 hr) and inulinase expression (1.0 units/mL with SEY2102/pYIGP and 2.5 units/mL with 2805/pYIGP) than other inulin sources such as dahlia and chicory. It was also found that maximal ethanol production of 9 g/L was obtained from Jerusalem artichoke inulin at the early stationary phase (around 30 hr), indicating that recombinant S. cerevisiae cells secreting exoinulinase could be used for the simultaneous saccharification of inulin and ethanol fermentation.

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Screenig and Indentification of Wild Strains for the Production of High Concentration of Alcohol from Jerusalem artichoke Tubers (돼지감자를 이용한 고농도 알코올발효 균주의 탐색)

  • Hong, Yeun;Choi, Eon-Ho
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.707-712
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    • 1994
  • Yeast screening for effective production of alcohol from Jerusalem artichoke tubers as an alternative energy source was performed. Inulin assimilative strains with high alcohol tolera- nce were isolated from wild sources and cultured in the liquid media of Jerusalem artichoke powder varying its concentraion from 15 to 30%. As a result, four strains of 2,445 isolates showing the inulin assimilation were selected as alcohol fermentative and alcohol tolerant yeasts. These strains were assignated to be Kluyveromyces marxianus F043 and Kluyveromyces sp. F173, E040, and F334, respectively, by their cultural and physiological characteristics. The F043 strain produced ethanol of 98.1 g/l in the 25% Jerusalem artichoke medium for 3 days.

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Secretory Overexpression of Clostridium Endoglucanase A in Saccharomyces cerevisiae Using GAL10 Promoter and Exoinulinase Signal Sequeice. (Saccharomyces cerevisiae에서 GAL 10 promoter와 exoinulinase 분비 서열을 이용한 Clostridium endoglucanase A의 과발현·분비)

  • Lim, Myung-Ye;Lee, Jin-Woo;Lee, Jae-Hyung;Kim, Yeon-Hee;Seo, Jin-Ho;Nam, Soo-Wan
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1248-1254
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    • 2007
  • The secretory overexpression of Clostridium thermocellum endoglucanase A gene (celA) was examined in Saccharomyces cerevisiae using Kluyveromyces marxianus exoinulinase (INU1) signal sequence and GAL10 promoter. The two plasmids, pYEG-CT1 with its own signal sequence, and pYInu-CT1 with INU1 signal sequence were introduced to S. cerevisiae SEY2102 and S. cerevisiae 2805 host strains, respectively, and then each transformant was selected on the synthetic defined media lacking uracil. The expression level and secretion efficiency of endoglucanase A was increased by $18{\sim}22%$ and 11%, respectively, by INU1 signal sequence over celA signal sequence. By considering the high level of expression (361 unit/I), plasmid stability (89%), and secretion efficiency (70%), S. cerevisiae 2805 harboring plasmid pYInu-CT1 was selected as the opti-mal host vector system for the production of cellulose-degrading enzyme and recombinant yeast probiotic. The total expression and secretion efficiency of endoglucanase A was 418 unit/l and 73%, respectively, in the batch fermentation of S. cerevisiae 2805/pYlnu-CT1 on galactose medium. The mo-lecular weight of secreted endoglucanase A was found to be greater than 100 kDa, presumably due to the N-linked glycosylation.

Occurrence of Yeasts in Cultivated Soils in El-Minia City, Egypt

  • Haridy, Mamdouh S.A.
    • Mycobiology
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    • v.30 no.1
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    • pp.27-30
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    • 2002
  • Two-hundred two yeast strains were isolated from rhizosphere(87 strains) and nonrhizosphere(115 strains) areas of potato, maize, vegetable marrow, and cabbage plants. On the basis of 26 morphological and physiological properties, the isolated yeast strains were assigned to 9 genera and 15 species. Trichosporon beigelii, Kluyveromyces marxianus and Torulaspora delbrueckii were the dominant species. Cryptococcus humicolus and Candida tropicalis were represented by considerable numbers of strains. Of low occurrence were Saccharomyces cerevisiae and Candida blankii. Other yeast species were represented by single or two strains. Total counts of yeast cells per gram dry soil ranged from $1.1{\times}10^3$ to $6.6{\times}10^3$ in soil samples of rhizosphere areas and from $6.5{\times}10^2$ to $5.6{\times}10^3$ in soil samples of nonrhizosphere areas. Types of the tested plants affected not only the total counts of yeast cells but also spectra of yeast species. Relationships of age of potato plant, moisture contests of soil samples, and its pH values and total counts of yeast cells were discussed.

Analysis of Microbial Diversity in Makgeolli Fermentation Using PCR-DGGE (PCR-DGGE를 이용한 막걸리발효에서 미생물 다양성 분석)

  • Kwon, Seung-Jik;Ahn, Tae-Young;Sohn, Jae-Hak
    • Journal of Life Science
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    • v.22 no.2
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    • pp.232-238
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    • 2012
  • Kumjungsansung-Makgeolli$^{(R)}$ is a traditional Korean rice wine that is fermented from traditional nuruk and rice. In this study, we performed the PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes to characterize bacterial and fungal diversity during Makgeolli fermentation. The predominant bacteria in the PCR-DGGE profile during Makgeolli fermentation were Lactobacillus spp. (Lactobacillus curvatus, L. kisonensis, L. plantarum, L. sakei, and L. gasseri), Pediococcus spp. (P. acidilactici, P. parvulus, P. agglomerans, and P. pentosaceus), Pantoea spp. (P. agglomerans and P. ananatis), and Citrobacter freundii; these were identified on the base of analysis of 16S rRNA gene sequences. The dominant bacterium during Makgeolli fermentation was L. curvatus. The predominant fungi in PCR-DGGE profile during Makgeolli fermentation were Pichia kudriavzevii, Saccharomyces cerevisiae, Asidia idahoensis, Kluyveromyces marxianus, Saccharomycopsis fibuligera, and Torulaspora delbrueckii, and these were identified on the basis of analysis of 28S rRNA gene sequences. The dominant fungal species during Makgeolli fermentation changed from P. kudriavzevii at 0-2 days incubation to S. cerevisiae at 3-6 days incubation. This study suggests that PCR-DGGE analysis could be a suitable tool for the understanding of microbial diversity and structure during Makgeolli fermentation.

Screening of Thermotolerant Yeast Strain for Ethanol Fermentation (Ethanol 발효를 위한 내열성 효모 균주의 Screening)

  • Ryu, Beung-Ho;Nam, Ki-Du;Kim, Hae-Sung;Kim, Dong-Seuk;Ji, Young-Ae;Jung, Soo-Ja
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.265-269
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    • 1988
  • For the purpose of developing new thermotolerant yeast strains for ethanol fermentation, yeasts were isolated from molasses and screened for their fermentation ability at elevated temperatures. Three candidate strains were screened. These strains preferred pH 5.0 and 34$^{\circ}C$ for their ethanol production. Under such conditions the three strains showed average ethanol productivity of 75g ethanol per liter of fermentation broth in n synthetic medium containing glucose as substrate. These strains were identified as Saccharomyces cerevisiae and Kluveromyces marxianus.

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Expression and Cloning of Microbial Transglutaminase in S. cerevisiae (세균 유래 단백질연결효소 Transglutaminase의 클로닝과 효모에서의 발현)

  • Kim, Hyoun-Young;Oh, Dong-Soon;Kim, Jong-Hwa
    • The Korean Journal of Mycology
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    • v.36 no.1
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    • pp.93-97
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    • 2008
  • A $Ca^{2+}-independent$ microbial transglutaminase (mTGase) from the actinomycete Streptomyces mobaraensis IFO13819 is a useful enzyme in the food industry. It is consists 406 amino acid residues, which comprised leader and pro region of 75 amino acid residues and the structure region of 331 amino acid residues. Pro and structure gene of TGase were cloned into the yeast shuttle vector pYAEG-TER and then used to transform Saccharomyces cerevisiae 2805. Expression of mTGase in recombinant was confirmed with Northern hybridization and the maximal activity of TGase was shown 26 mU/ml.

Influence of Nutrient Addition in the Liquid Yeast Fermentation of Pulverized Food Wastes (남은 음식물의 습식효모배양에서 영양물질첨가가 효모증식에 미치는 영향)

  • Lee, Ki-Young;Yu, Sung-Jin;Chae, Hee-Jung
    • Journal of the Korea Organic Resources Recycling Association
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    • v.9 no.1
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    • pp.49-55
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    • 2001
  • For the production of probiotic feed enriched with viable yeasts, aerobic liquid culture of Kluyveromyces marxianus was attempted in pulverized residual food wastes. After the preliminary shaking culture result, the liquid food wastes was added with urea($0.5g/{\ell}$), o-phosphate($0.4g/{\ell}$ ), molasses($4g/{\ell}$), and yeast extract($1g/{\ell}$), and the fermentation was carried out in 2-litre jar fermenter. In 12 hours of aerobic mixed culture with Aspersillus oryzae, viable cell count of the yeast reached to the number of $1.4{\times}10^{10}/{\ell}$ in the cultured medium.

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Cloning and Characterization of the Orotidine-5'-Phosphate Decarboxylase Gene (URA3) from the Osmotolerant Yeast Candida magnoliae

  • Park, Eun-Hee;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.642-648
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    • 2012
  • We determined the nucleotide sequence of the URA3 gene encoding orotidine-5'-phosphate decarboxylase (OMPDCase) of the erythritol-producing osmotolerant yeast Candida magnoliae by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed the presence of an uninterrupted open-reading frame of 795 bp, encoding a 264 amino acid residue protein with the highest identity to the OMPDCase of the yeast Kluyveromyces marxianus. Phylogenetic analysis of the deduced amino acid sequence revealed that it shared a high degree of identity with other yeast OMPDCase homologs. The cloned URA3 gene successfully complemented the ura3 null mutation in Saccharomyces cerevisiae, revealing that it encodes a functional OMPDCase in C. magnoliae. An enzyme activity assay and reverse transcription polymerase chain reaction indicated that the expression level of the C. magnoliae URA3 gene in S. cerevisiae was not as high as that of the S. cerevisiae URA3 gene. The GenBank accession number for C. magnoliae URA3 is JF521441.

Screening and Characterization of Thermotolerant Alcohol-producing Yeast

  • Sohn, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.215-221
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    • 1994
  • Two strains of yeast (RA-74-2 and RA-912) showing superior fermenting ability at a high temperature were isolated from soils and wastewaters by an enrichment culture method. Based on the morphological and physiological charateristics, the two strains were identified as Saccharomyces cerevisiae and Kluyveromyces marxianus, respectively. RA-74-2 was able to grow upto $43^{\circ}C$ and sustain similar fermenting ability in the temperatures range from 30 to $40^{\circ}C$. In addition, the sugar- and ethanol-tolerance of RA-74-2 were 30% (w/v) glucose and 10% (v/v) ethanol, which appeared to be higher than those of nine other industrial yeast strains currently being used in the alcohol factories. The thermotolerant ethanol fermenting yeast RA-912 showed identical growth in the temperatures range from 35 to $45^{\circ}C$ and was resistant to various heavy metals. The quality and quantity of byproducts of the isolated yeast strains in fermentation broth after fermentation at $40^{\circ}C$ and $45^{\circ}C$ were similiar with those obtained at $30^{\circ}C$. These results show that RA-74-2 can be adopted for the ethanol fermentation process where the expenses for cooling system is significant, and suggest that RA-912 may be applied in either SSF(simultaneous saccharification and fermentation) or Flash-fermentation process and RA-912 may be used as a gene donor for the development of thermotolerant ethanol-fermenting yeasts.

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