Molecular Cloning and Expression of a Cellulolytic Xylanase Gene from Bacillus circulans in Escherichia coli

Bacillus circulans 기원의 Cellulolytic Xylanase 유전자의 대장균에서의 클로닝 및 발현

  • 이동석 (인제대학교 의생명공학대학 임상병리학과) ;
  • 김지연 (인제대학교 생물건강산업유성지원 센터) ;
  • 김한복 (호서대학교 자연과학부 생명과학전공)
  • Published : 2000.09.01

Abstract

A gene for cellulolytic xylanase of Bacillus circulnns ATCC21365 was cloned on pUC 19 in Eschwichia coli. The recombinant plasniid pXLI80 contained an 1.8 id, inselt composed of0.5 kb and 1.3 kb PslI fragments derived from B, circulans. The 0.5 kh fragment in the upstream region of 1.3 kb one was confirmed lo be indispensable for not only expression but also hyperexpression of the cloned gene. The transformant overproduced the xylanase 135 times greater than that produced by the orlginal B circulnns. The optimum pH and temperature of the cloned enzyme we]-e pH 5.2 and $60^{\circ}C$, respectively. Heal pretl-eatment at TEX>$55^{\circ}C$C for 1 Indid not cause inhibition of the activity of this enzyme. The elm.ynie could hydl-olyre CMC and lichenan as well as xylan to produce xylose(or GI), xylohiose(or G2) and xylolnose(or G3) as inah products. Hence We defined the cloned enzyme as a cellulolytic xylanase. The SDS-PAG electrophoretic mobility and zyiiogram of this enzyme derived from whole cell extracts or c~~lture supematants or E. coli(pXL180) indicated a molecular weight of 45,000 and nonprocessing of the enzyme in the peilplasln of E. coli.

Keywords

Bacillus circulans;cellulolytic xylanase;cloning

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