Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Properties of Pectate Lyase Produced by Recombinant Strain -Containing pelK Gene from Alkalitolerant Bacillus sp. YA- 14

알칼리내성 Bacillus sp. YA-14 유래의 Pectate Lyase 유전자를 함유한 재조합균주로부터 효소의 정제 및 특성

  • 한혜정 (연세대학교 식품공학과) ;
  • 김진만 (연세대학교 식품공학과) ;
  • 박희경 (연세대학교 식품공학과) ;
  • 배동훈 (단국대학교 식품공학과) ;
  • 유주현 (연세대학교 식품공학과)
  • Published : 1992.12.01
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Abstract

Pectate lyase produced by recombinant strain containing pectate lyase gene from alkalitolerant Bacillus sp. YA-14 was succesively purified with 257.6 purification folds and a 10.2% yields by the affinity method, eM-cellulose column chromatography followed by gel filtration on Sephadex G-I00 column. The optimal pH and temperature for pectate lyase activity were 10.0 and $60^{\circ}C$, respectively. The enzyme was stable between pH 4.0 and 10.0, and up to $50^{\circ}C$. The molecular weight of this enzyme was estimated to be 43,000 daltons by SDS-PAGE. Amino acid analysis showed that the enzyme contained more polar and basic amino acids, especially serine, glycine and tyrosine, than that of various pectate lyase from other strains. The N-terminal amino acid sequence was Ala-Asp-Leu-Gly-His-Gln-Thr.

알칼리내성 Bacillus sp. Ya-14 유래의 pectate lyase 유전자를 함유한 재조합균주로부터 affinity method, CM-cellose column chromatography와 gel filtration을 통해 효소를 정제하였으며 정제효소의 수율은 10.2, 정제도는 258배였다. 효소의 최적활성 pH는 10.0이었고 pH4.0-10.0까지의 범위에서 안정성이 있었으며, 최적활성온도는 $60^{\circ}C$이고 $50^{\circ}C$까지 열안정성이 있으며, SDS-PASGE에 의해 추정된 분자량은 43KDa 이었다. 아미노산 조정 분석 결과 polar, basic 아미노산의 함량이 높고 특히 Ser, Gly, Tyr의 함량이 높았으며, 정제효소의 N-terminal은 Ala-Asp-Leu-Gly-His-Gln-Thr의 아미노산 서열이었다.

Keywords

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