Bacillus megaterium KSM B-404으로부터 생산되는 Maltopentaose생성 Amylase의 정제 및 특성

Purification and Characterization of a Maltopentaose-producing Amylase from Bacillus megaterium KSM B-404.

  • 박제원 (고려대학교 생명공학원) ;
  • 김병주 (㈜태평양 기술연구원) ;
  • 이재우 (김천대학 식품영양과) ;
  • 김영배 (고려대학교 생명공학원)
  • 발행 : 2002.12.01

초록

토양으로부터 maltopentaose생산성 amylase를 분비하는 세균 KSM B-404를 분리하여 그의 형태적, 생리적인 특성을 고려한 결과 Bacillus megaterium으로 동정되었다. 효소는 ($NH_4$)$_2$$SO_4$ 침전 분획, DEAE-Toyopearl 및 Superdex 75 HR 10/30 크로마토그래피로 129배 정제되었으며 21.4%의 활성이 회수되었다. 정제된 효소를 SDS-PACE로 분석한 결과 분자량은 약 68 kDa이었고, 최적 반응 온도는 $50^{\circ}C$이며 $Ca^{2+}$ 이온의 존재 시 열 안정성이 증가하였다. 한편 최적 반응 pH는 6.0~7.0부근이며 알칼리 조건에서도 안정하였다. 또한 효소의 활성은 $Cu^{2+}$ , $Hg^{2+}$ 그리고 특히 Fe/eup 3+/이온 등의 금속이온에 의해 강하게 저해 받았고 acetic anhydride, EDTA , hydroxylamine-HCI, $\rho$ - chloromercuribenzoate 등의 저해제에 의해서 활성이 저해되었으나 concanavalin A에 의한 저해 효과는 나타나지 않았다. 전분의 가수분해 산물을 HPLC로 분석한 결과 maltopentaose가 주산물로 나타났으며 반응 24시간 후 총 가수분해 산물의 약 52%를 차지하였다.

키워드

Maltopentaose;Bacillus megaterium KSM B-404;amylase

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