Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Isolation, Purification and Characterization of the β-Xylosidase from Klebsiella sp. Sc.

Klebsiella sp. Sc가 생산하는 β-xylosidase의 분리, 정제 및 특성

  • Lee, Yong-Seok (Department of Biotechnology, College of Natural Resources and Life Science, Dong-a University) ;
  • Park, In-Hye (Department of Biotechnology, College of Natural Resources and Life Science, Dong-a University) ;
  • Ahn, Soon-Cheol (Department of Microbiology and Immunlogy, Pusan National University school of Medicine) ;
  • Choi, Yong-Lark (Department of Biotechnology, College of Natural Resources and Life Science, Dong-a University)
  • Received : 2010.11.26
  • Accepted : 2010.12.08
  • Published : 2010.12.30
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Abstract

A $\beta$-xylosidase encoding gene from Klebsiella sp. Sc was cloned in Escherichia coli. The $\beta$-xylosidase gene consisted of an open reading frame of 1,680 nucleotides and encodes 559 amino acids with a deduced molecular weight of 63 kDa. The deduced amino acid sequence of the $\beta$-xylosidase from Klebsiella sp. Sc exhibits 90% identities and 95% positives compared to those from Klebsiella oxytoca (KOX), Lactobacillus lactis (LAC, 82%, 90%), Bacillus longum (BLON, 69%, 81%) and Escherichia coli (ECOLI, 47%, 63%). The $\beta$-xylosidase was purified by GST-fusion purification system. The pH and temperature optima of the enzyme were 6.6 and $55^{\circ}C$, respectively. The $\beta$-xylosidase hydrolyzes xylobiose to xylose.

Klebsiella sp. Sc로부터 birchwood xylan을 분해하는 $\beta$-xylosidase를 분리하였다. 이 $\beta$-xylosidase는 63 kDa의 분자량을 가지는 559개의 아미노산을 암호화하며 1,680개의 뉴클레오타이드로 구성 되는 것으로 밝혀졌다. 기존에 밝혀진 세균성 $\beta$-xylosidase와 상동성을 비교해 보았을 때, Klebsiella oxytoca (KOX)와 90% identities와 95% positives를 나타내었으며 Lactobacillus lactis (LAC, 82%, 90%), Bacillus longum (BLON, 69%, 81%) 그리고 Escherichia coli (ECOLI, 47%, 63%)를 나타내었다. 분리된 $\beta$-xylosidase는 GST-fusion 정제 시스템을 이용하여 순수 정제하였다. 이 효소 활성의 최적 pH는 6.6이었으면 최적 온도는 $55^{\circ}C$였다. TLC를 통해 효소 분해 산물을 관찰한 결과, xylobiose를 분해하여 xylose를 생산하는 것을 관찰 할 수 있었다.

Keywords

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