• Title/Summary/Keyword: Schwanniomyces occidentalis glucoamylase gene

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Cloning and Expression of a Paenibacillus sp. Neopullulanase Gene in Saccharomyces cerevisiae Producing Schwanniomyces occidentalis Glucoamylase

  • Kim, Hyo-Jeong;Park, Jeong-Nam;Kim, Hee-Ok;Shin, Dong-Jun;Chin, Jong-Eon;Blaise Lee, Hwang-Hee;Chun, Soon-Bai;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.340-344
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    • 2002
  • A gene, npl, encoding neopullulanase from Paenibacillus sp. KCTC 8848P was cloned and expressed in Escherichia coli. It consisted of an open reading frame of 1,530 bp for a protein that consisted of 510 amino acids with a molecular weight of 58,075 Da. The deduced amino acid sequence of the neopullulanase gene had $92\%$ identity with the neopullulanase of Bacillus polymyxa. The npl gene was also expressed in Saccharomyces cerevisiae secreting Schwanniomyces occidentalis glucoamylase (GAM1) under the control of the yeast actin gene (ACT1) promoter. Secretion of the neopullulanase was directed by the yeast mating pheromone ${\alpha}$ -factor ($MF{\alpha}1$) prepro region. Enzyme assays confirmed that co-expression of npl and GAM1 enhanced starch and pullulan degradation by S. cerevisiae.

Expression of Schwanniomyces occidentalis $\alpha-Amylase$ Gene in Saccharomyces cerevisiae var. diastaticus

  • Park, Jeong-Nam;Shin, Dong-Jun;Kim, Hee-Ok;Kim, Dong-Ho;Lee, Hwang-Hee;Chun, Soon-Bai;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.668-671
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    • 1999
  • The gene encoding Schwanniomyces occidentalis $\alpha-amylase$(AMY) was introduced into Saccharomyces cerevisiae var. diastaticus which secreted only glucoamylase, by using a linearized yeast integrating vector to develop stable strains with a capability of secreting $\alpha-amylase$and glucoamylase simultaneously. A dominant selectable marker, the geneticin(G418) resistance gene (Gt^r$), was cloned into a vector to screen wild-type diploid transformants harboring the AMY gene. The amylolytic activities of transformants were about 3-7 times higher than those of the recipient strains. When grown in nonselective media, the transformants with the linearized integrating vector containing the AMY gene exhibited almost all of the mitotic stability after 100 generations.

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