Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Expression of Aspergillus awamori Glucoamylase Gene in an Industrial Strain of Saccharomyces cerevisiae

산업용 Saccharomyces cerevisiae에서 Aspergillus awamori Glucoamylase 유전자의 발현

  • Ghang Dong-Myeong (Department of Biological Sciences, Institute of Resources Plant, Chonnam National University) ;
  • Lee Su-A (Department of Biological Sciences, Institute of Resources Plant, Chonnam National University) ;
  • Chun Young-Hyun (Department of Biological Sciences, Institute of Resources Plant, Chonnam National University) ;
  • Chin Jong-Eon (Department of Cosmetology, Dongkang College) ;
  • Lee Hwanghee Blaise (Department of Biological Sciences, Institute of Resources Plant, Chonnam National University) ;
  • Bai Suk (Department of Biological Sciences, Institute of Resources Plant, Chonnam National University)
  • 강동명 (전남대학교 자원식물연구소 생물학과) ;
  • 이수아 (전남대학교 자원식물연구소 생물학과) ;
  • 전영현 (전남대학교 자원식물연구소 생물학과) ;
  • 진종언 (동강대학 피부미용과) ;
  • 이황희 (전남대학교 자원식물연구소 생물학과) ;
  • 배석 (전남대학교 자원식물연구소 생물학과)
  • Published : 2005.06.01
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Abstract

To construct an amylolytic industrial strain of Saccharomyces cerevisiae, the glucoamylase cDNA gene (GAl) from Aspergillus awamori was expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) and integrated into the chromosomes of industrial S. cerevisiae. An integrative cassette lacking bacterial ampicillin resistance gene but containing the GA1 gene, $\delta$ sequences of Ty1 retrotransposon as target sites for homologous recombination and S. cerevisiae aureobasidin A resistance gene (AUR1-C) as the selection marker was constructed to obtain a strain eligible for commercial use. Industrial S. cerevisiae transformed with this 15-integrative cassette efficiently secreted glucoamylase into the medium and grew on starch as the sole carbon source. The transformants were mitotically stable for 100 generations in nonselective medium.

전분 이용이 가능한 산업용 Saccharomyces cerevisiae균주를 개발하기 위해 alcohol dehydrogenase 유전자 프로모터(ADClp)의 조절하에 발현되는 Aspergillus awamori glucoamylase cDNA 유전자(GA1)를 산업용 S. cerevisiae의 염색체에 도입하였다. 산업적 이용에 적합한 효모균주를 얻기 위해 세균 ampicillin 저항성 유전자가 제거되고 GA1 유전자와 선별 표지유전자로 S. cerevisiae aureobasidin A 저항성 유전자(AUR1-C)와 재조합 부위로 Tyretrotransposon $\delta$-서열이 포함된 integrative cassette를제조하였다. 이 $\delta-integrative$ cassette로 형질전환된 산업용 S. cerevisiae는 배지상에 glucoamylase를 생산 분비하였고 전환을 유일한 탄소원으로 하여 생장하였다. 형질전환체를 비선택배지에서 배양했을 매 삽입된 GA1유전자가 100세대까지 안정되게 유지되었다.

Keywords

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