한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제37권2호
  • /
  • Pages.147-152
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  • 2009
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  • 1598-642X(pISSN)
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  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Bacillus sp. A-6의 Xylanase의 정제와 특성

Purification and Characterization of Xylanase from Bacillus sp. A-6

  • 최석호 (상지대학교 동물생명자원학부)
  • Choi, Suk-Ho (Division of Animal Science and Life Resources, Sangji University)
  • 투고 : 2009.03.02
  • 심사 : 2009.04.16
  • 발행 : 2009.06.28
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초록

Bacillus sp. A-6의 배양액의 상등액으로부터 한외여과와 5 mM sodium acetate, pH 5.0 용액으로 평형화된 SP-Sepharose column을 사용한 이온교환 크로마토그래피에 의해 xylanase를 정제하였다. Column에 흡착된 xylanase는 0.05 M NaCl 이하의 농도에서 용출되었다. 용출된 xylanase가 SDS-PAGE에서 단일 펩티드 밴드로 분리되어 순수함을 확인하였으며 oat spelt xylan을 기질로한 zymogram에서 xylan을 분해하는 밴드로 나타났다. Xylanase의 분자량은 SDS-PAGE에서 15,000이었고 겔여과 크로마토그래피에서 14,100 이었다. 박층막 크로마토그래피에서 xylanase가 oat spelt xylan을 xylobiose와 xylooligosaccharide로 분해함을 보였다. Xylanase를 가열할 때에 상대활성도가 $40^{\circ}C$에서 7시간 후에 80%로 감소하였으며 $60^{\circ}C$에서는 1시간 후에 40% 이하로 감소하였다.

A xylanase was purified from the culture supernatant of Bacillus sp. A-6 by using ultrafiltration and ion exchange chromatography on the column of SP-Sepharose using 5 mM acetate buffer, pH 5.0. The xylanase was eluted from the column at the concentration less than 0.05 M NaCl. The eluted xylanase was shown to be a single protein band in SDS-PAGE. Zymogram analysis indicated that the protein band in SDS-PAGE had the enzyme activity to hydrolyze oat spelt xylan. The molecular weights of the xylanase were 15,000 based on SDS-PAGE and 14,100 based on gel filtration chromatography. Thin layer chromatography showed that the xylanase hydrolyzed oat spelt xylan into xylobiose and high-molecular-weight xylooligosaccharides. The relative activities of the heated xylanase decreased to 80% at $40^{\circ}C$ after 7 hr and less than 40% at $60^{\circ}C$ after 1 hr.

키워드

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