Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Expression of a Paenibacillus sp. Neopullulanase Gene in Saccharomyces cerevisiae Producing Schwanniomyces occidentalis Glucoamylase

  • Kim, Hyo-Jeong (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Park, Jeong-Nam (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Kim, Hee-Ok (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Shin, Dong-Jun (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Chin, Jong-Eon (Department of Cosmetology, DongKang College) ;
  • Blaise Lee, Hwang-Hee (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Chun, Soon-Bai (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University) ;
  • Bai, Suk (Department of Biological Sciences, The Institute of Basic Sciences, Chonnam National University)
  • Published : 2002.04.01
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Abstract

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.

Keywords

References

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