Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Isolation and biochemical characterization of acid tolerance xylanase producing Bacteria, Bacillus sp. GJY from city park soil

도심공원으로부터 산내성 xylanase를 생산하는 박테리아 분리 및 효소학적 특성

  • Jang, Min-Young (Department of Forest Resources, Gyeongnam National University of Science and Technology) ;
  • Park, Hwa Rang (Department of Forest Resources, Gyeongnam National University of Science and Technology) ;
  • Lee, Chong Gyu (Department of Forest Resources, Gyeongnam National University of Science and Technology) ;
  • Choo, Gab-Chul (Department of Forest Resources, Gyeongnam National University of Science and Technology) ;
  • Cho, Hyun Seo (Department of Forest Resources, Gyeongnam National University of Science and Technology) ;
  • Park, Sam-Bong (Department of Forest Resources, Gyeongnam National University of Science and Technology) ;
  • Oh, Ki-Cheol (Nakdong River Basin Environmental Office) ;
  • Kim, Bong-Gyu (Department of Forest Resources, Gyeongnam National University of Science and Technology)
  • Received : 2017.01.23
  • Accepted : 2017.01.25
  • Published : 2017.03.30
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Abstract

Microbes in forest are very important due to not only to enhance soil fertility but also maintain a healthy ecosystem by supplying the energy available to living organisms by producing various kinds of enzymes related to degradation of lignocellulosic biomass. In order to isolate a lignocellulosic biomass degrading bacterial strain from the Jurassic park located in Gyeongnam National University of Science and Technology, We used the Luria-Bertani-Carboxymethyl cellulose (CMC) agar trypan blue method containing 0.4 % carboxymethyl cellulose and 0.01 % trypan blue. As a result, we isolated a bacterial strain showing both activity on the CMC and xylan. To identify the isolated strain, 16S rRNA sequencing and API kit analysis were used. The isolated strain turned out to belong to Bacillus species and then named Bacillus sp. GJY. In the CMC zymogram analysis, it showed that one active band of about 28kDa in size is present. Xylan zymogram analysis also showed to have one active band of about 25kDa in size. The optimal growth temperature of Bacillus sp. GJY was $37^{\circ}C$. The maximal activities of CMCase and xylanase were 12 hour after incubation. The optimal pH and temperature for CMCase were 5.0 and $40^{\circ}C$, respectively, whereas the optimal pH and temperature for xylanase was 4.0 and $40^{\circ}C$. Both activities for CMCase and xylanase showed to be thermally stable at 40and $50^{\circ}C$, while both activities rapidly decreased at over $60^{\circ}C$.

토양 내에 존재하는 미생물은 다양한 종류의 다당류 분해 효소들을 생산함으로써 토양의 비옥도 증진뿐만 아니라 토양내의 생태계를 건전하게 유지하게 위해 중요한 역할을 한다. 본 연구는 경남과학기술대학교에 위치한 쥬라기숲에서 0.4 % carboxymethyl cellulose와 0.01 % trypan blue가 첨가된 LB agar plate를 이용하여 CMCase와 xylanase를 생산하는 박테리아를 분리하였다. 16S rRNA 유전자 염기서열 분석과 API kit 분석을 바탕으로 분리된 박테리아는 Bacillus 종에 속하는 것으로 동정되었으며, Bacillus sp. GJY으로 명명하였다. Bacillus sp. GJY에서 CMCase와 xylanase의 활성을 책임지고 있는 단백질을 알아보기 위하여 Zymogram 분석을 실시하였다. 그 결과 CMCase의 경우 약 28 kDa 크기에 xylanase의 경우 약 25 kDa 크기에 활성밴드가 하나씩 존재하였다. Bacillus sp. GJY의 최적 생장온도는 $37^{\circ}C$이었으며, CMCase와 xylanase의 활성은 배양 후 12시간에 최고에 달하였다. CMCase의 경우 pH 5.0, 온도 $40^{\circ}C$에서 최적의 활성을 보인 반면, xylanase는 pH 4.0, $40^{\circ}C$에서 최적의 활성을 보였다. CMCase와 xylanase 모두 40, $50^{\circ}C$에서는 열 안정성을 보였지만, $60^{\circ}C$ 이상에서는 두 효소의 열 안정성이 급격하게 감소하는 경향을 보였다.

Keywords

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