• Title/Summary/Keyword: Pichia media

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A New Isolation and Evaluation Method for Marine-Derived Yeast spp. with Potential Applications in Industrial Biotechnology

  • Zaky, Abdelrahman Saleh;Greetham, Darren;Louis, Edward J.;Tucker, Greg A.;Du, Chenyu
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1891-1907
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    • 2016
  • Yeasts that are present in marine environments have evolved to survive hostile environments that are characterized by high exogenous salt content, high concentrations of inhibitory compounds, and low soluble carbon and nitrogen levels. Therefore, yeasts isolated from marine environments could have interesting characteristics for industrial applications. However, the application of marine yeast in research or industry is currently very limited owing to the lack of a suitable isolation method. Current methods for isolation suffer from fungal interference and/or low number of yeast isolates. In this paper, an efficient and non-laborious isolation method has been developed and successfully isolated large numbers of yeasts without bacterial or fungal growth. The new method includes a three-cycle enrichment step followed by an isolation step and a confirmation step. Using this method, 116 marine yeast strains were isolated from 14 marine samples collected in the UK, Egypt, and the USA. These strains were further evaluated for the utilization of fermentable sugars (glucose, xylose, mannitol, and galactose) using a phenotypic microarray assay. Seventeen strains with higher sugar utilization capacity than the reference terrestrial yeast Saccharomyces cerevisiae NCYC 2592 were selected for identification by sequencing of the ITS and D1/D2 domains. These strains belonged to six species: S. cerevisiae, Candida tropicalis, Candida viswanathii, Wickerhamomyces anomalus, Candida glabrata, and Pichia kudriavzevii. The ability of these strains for improved sugar utilization using seawater-based media was confirmed and, therefore, they could potentially be utilized in fermentations using marine biomass in seawater media, particularly for the production of bioethanol and other biochemical products.

Isolation of Wild Yeasts from the Water and Riverside Soil of Geumgang Midstream in Sejong City, Korea, and Characterization of Unrecorded Wild Yeasts (세종특별자치시 주변의 금강 중류 물과 토양에서 야생 효모의 분리 및 국내 미기록 효모의 특성)

  • Han, Sang-Min;Kim, Ji-Yoon;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.51-61
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    • 2019
  • The goal of this study was to elucidate wild yeast diversity of Geumgang midstream near Sejong metropolitan autonomous city, Korea. Thirty-seven strains of 32 species of wild yeasts were isolated from 43 water and soil samples under the Bulti bridge of Sejong city, Korea. Seven yeasts of each Candida spp. and Cryptococcus spp. were the predominant species isolated from samples near the Bulti bridge. Holtermanniella takashimae SW048 (NNIBRFG9314), Cystofilobasidium infirmominiatum SW013 (NNIBRFG9310), Mrakia cryoconite SW015 (NNIBRFG9316), Pichia sporocuriosa SW085 (NNIBRFG9326) and Cryptococcus aspenensis SW008 (NNIBRFG9309) represented novel yeast strains found in Korea for the first time. All of these previously unrecorded yeasts, except for Mrakia cryoconite SW015 had ascospores and grew well in yeast extract-peptone-dextrose (YPD), yeast extract-malt extract (YM) and potato-extrose (PD) media. Pichia sporocuriosa SW085 grew well in vitamin-free medium and Holtermanniella takashimae SW048, which was a halotolerant wild yeast, grew well YPD medium containing 5 % NaCl. Twenty-six strains representing eight species of wild yeast were isolated from 22 water and soil samples under the Haetmuri bridge of Sejong city, Korea. Candida pseudolambica (12 strains) and Aureobasidium pullulans (11 strains) were the predominant isolates from samples near the Haetmuri bridge. Occultifur kilbournensis HB060 (NNIBRFG9317), Sampaiozyma vanillica HB014 (NNIBRFG9332), Xenoramularia neerlandica HB039 (NNIBRFG9335), Candida norvegica HB315 (NNIBRFG9306), C. melibiosica HB316 (NNIBRFG9305), C. quercuum GB014 (NNIBRFG9307), and C. succiphila GB015 (NNIBRFG9308) represented novel yeast strains recorded in Korea for the first time. O. kilbournensis HB060 and X. neerlandica HB039 did not form ascospores or pseudo-mycelia. All of these previously unrecorded yeasts, except S. vanillica HB014 and X. neerlandica HB039, grew well in vitaminfree medium, and C. norvegica HB315 and C. succiphila GB015, which were halotolerant wild yeasts, which grew well in YPD medium containing 5 % NaCl.

Enhanced Production of Succinic Acid by Actinobacillus succinogenes using the Production Medium Supplemented with Recombinant Carbonic Anhydrases (재조합 탄산무수화 효소 첨가 생산배지를 이용한 Actinobacillus succinogenes 유래의 숙신산 생산성 향상)

  • Park, Sang-Min;Eum, Kyuri;Kim, Sangyong;Jeong, Yong-Seob;Lee, Dohoon;Chun, Gie-Taek
    • KSBB Journal
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    • v.29 no.3
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    • pp.155-164
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    • 2014
  • Succinic acid, a representative biomass-derived platform chemical, is a major fermentation product of Actinobacillus succinogenes. It is well known that carbon dioxide is consumed during the succinate fermentation, but the biochemical mechanism behind this phenomenon is not yet understood well. In this study, it was found that the addition of carbonic anhydrase (CA)s into media significantly enhances the succinic acid production by A. succinogenes during the fermentation supplied with carbon dioxide. It is likely that the (bi) carbonate produced by the CA activity from gaseous carbon dioxide is favoured by A. succinogenes for consumption and utilization. Therefore, the $MgCO_3$ requirement could be significantly reduced without compromising the succinate productivity. Furthermore, because of too high price of the commercial carbonic anhydrase, it was undertaken to economically overproduce a cyanobacterial carbonic anhydrase by the use of a recombinant Pichia pastoris. An expression vector system was constructed with the carbonic anhydrase gene PCR-cloned from Cyanobacterium Synechocystis sp., and introduced into P. pastoris for fermentation studies. About 95.9 g/L of succinic acid was produced in the production medium with 30 ppm of carbonic anhydrase, approximately 2 fold higher productivity compared to the parallel process with no supplementation of the enzyme. It is expected that this method can provide a valuable way of overcoming inefficiencies inherent in gas supply during $CO_2$-based bioprocesses like succinic acid fermentation.

Characteristics of wild yeast isolated from non-sterilized Makgeolli in Korea (국내 생막걸리에서 분리한 야생 효모의 특성)

  • Jung, Su Ji;Yeo, Soo-Hwan;Mun, Ji-Young;Choi, Han-Seok;Baek, Seong Yeol
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.1043-1051
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    • 2017
  • Wild yeasts were isolated from domestic non-sterilized Makgeolli and their fermentation characteristics were analyzed to select the best fermentation seed culture. A total of 65 yeast strains isolated yeasts from non-sterilized Makgeolli and Nuruk. In order to select fermentable strains, hydrogen sulfide, $CO_2$ production ability, alcohol tolerance and aroma component production ability were analyzed. To screen the aromatic strains of isolates, media containing cerulenin, 5,5,5-trifluor-DL-leucine (TFL) and API ZYM kit were used. There were 36 strains resistance to cerulenin and all strains produced esterase and demonstrated tolerance against TFL. Hydrogen sulfide, which could degrade the quality of the fermented beverage, was not produced in 34 yeast. The correlation between alcohol tolerance of yeast and carbon dioxide production was analyzed by principal component analysis. YM22, YM31, YM32 and YM37 produced a total of 0.14-0.18 g/72 h of $CO_2$ indicating high fermentability. Alcohol tolerance was measured by alcohol concentration. YM32, YM37 yeast had 20% alcohol tolerance. As a result, alcohol and flavor characteristics of wild yeast isolated from non-sterilized Makgeolli were analyzed and it was confirmed that yeast was suitable for the production of alcohol.