Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Expression Pattern of Acetyl Xylan Esterase of Streptomyces coelicolor A3(2) in Escherichia coli

Escherichia coli에서의 Streptomyces coelicolor A3(2)의 acetyl xylan esterase 발현 양상

  • 이인숙 (단국대학교 미생물학전공) ;
  • 윤석원 (단국대학교 미생물학전공) ;
  • 정상운 (단국대학교 의학레이저연구센터) ;
  • 오충훈 (단국대학교 의학레이저연구센터) ;
  • 김재헌 (단국대학교 의학레이저연구센터)
  • Published : 2003.06.01
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Abstract

We cloned a gene encoding acetyl xylan esterase(axeA) of Streptomyces coelicolor A3(2) and studied its expression pattern in Escherichia coli. The full sequence of axeA was amplified by PCR. Sequence analysis of the PCR product revealed an open reading frame of 1,008 nucleotides encoding a protein consisted of 335 amino acid residues, with a calculated molecular mass of about 38 kDa. The base sequence showed 98% homology to the same gene of Streptomyces lividans. Two different kinds of acetyl xylan esterases were produced in Escherichia coli(pLacI) by IPTG induction; their molecular weights were 38 kDa and 34 kDa, respectively. Of these, 38 kDa protein seemed to be a total protein holding N-terminal signal peptide region, whereas 34 kDa protein seemed to be a matured protein without signal peptide which was produced by peptide bond cleavage between two amino acid residues of alanine 41 and alanine 42.

streptomyces coelicolor A3(2)의 acetyl xylan esterase 유전자를 Escherichia coli께 클로닝하여 그 발현양상을 조사하였다. 이를 위하여 유전자 전체 DNA를 PCR증폭을 통하여 제조하였다. PCR산물의 염기서열을 분석한 결과 1,008개의 nucleotide로 구성된 하나의 open reading frame이 존재함을 확인하였고, 이것은 335개의 아미노산으로 이루어진 약 38 kDa의 단백질을 생성할 것으로 예측할 수 있었다. 이 유전자의 염기 서열은 streptomyces lividans의 acetyl xylan esterase와 98%의 상동성을 가졌다. 그런데 Escherichia coli (pLacl)에서 IPTG유도에 의해 두가지의 acetyl xylan esterase가 발현되었으며 각각의 분자량은 38 kDa과 34 kDa이었다. 이중에서 38 kDa의 단백질은 N-말단의 signal peptide를 포함한 전체 단백질이고,34 kDa의 단백질은 41번과 42번의 두 알라닌 잔기사이의 펩티드 결합이 끊어져 생산된 것으로 추정되었다.

Keywords

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