• Applied Biological Chemistry
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• 제50권2호
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• pp.107-110
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• 2007

일반적 대장균 배지 조건에서는 발현되지 않는 대장균 K-12의 laccase gene(yacK)을 PCR로 증폭한 후 pET28c에 클로닝하여 과량 발현시켰다. 과량 생산된 laccase를 His-affinity 칼럼 크로마토그래피로 정제하였다. SDS-PAGE 방법으로 확인한 과량 발현된 단백질의 분자량은 약 55,000이었으며, guaiacol 용액과 agar 배지에서 역가를 보여주었고 최적 온도는 65$^{\circ}C$, 최적 pH는 5이었다.

• 산업기술연구
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• 제37권1호
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• pp.16-20
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• 2017

A new putative laccase gene (soncotA) which show 78% homology with that from Bacillus licheniformis (liccotA) was isolated from draft genome sequence of Bacillus sonorensis KCTC 13918. A 1,545 bp of PCR product corresponding 514 amino acids was cloned into NdeI-NotI site of pET21c and expressed as soluble form in E. coli. About 59 kDa size of recombinant laccase was purified into homogenity by Ni-NTA column and laccase activity was confirmed by zymography. The enzymatic properties of recombinant laccase were characterized. The specific activity of B. sonorensis laccase was 0.033 fold lower than that of Bacillus licheniformis laccase. The finding of new laccase gene broadened the enzymatic diversity of Bacillus species laccases.