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《Chinese Journal of Biotechnology》 2007-06
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High Output of a Trametes Laccase in Pichia pastoris and Characterization of Recombinant Enzymes

ZHOU Hong-Min~ 1 , HONG Yu-Zhi~ 1 , XIAO Ya-Zhong~ 1 , CUI Teng-Jiao~2, WANG Xiao-Tang~2 andPU Chun-Lei~31 School of Life Sciences & Modern Experiment Technology Center, Anhui University, Hefei 230039, China2 Department of Chemistry and Biochemistry, Florida International University, 11200 SW 8th Street, Miami, FL 33199, USA3 School of Life Science, University of Science and Technology of China, Hefei 230026, China  
A laccase gene (lacD) from the basidiomycete Trametes sp. 420 was heterologously expressed in Pichia pastoris in two ways, resulting in two recombinant enzymes of rLacDx with native N-terminus and rLacDe with eight additional amino acid residues at N-terminus. The yields of rLacDx and rLacDe in shaken-flask cultures after an 18-day growth were 1.21×10~5u/L and 7.38×10~4u/L, respectively, as determined with 2,2'-azinobis(3-ethylbenzothia-zoline-6-sulfonic acid) (ABTS) as substrate. The yield of rLacDx was further increased to 2.39×10~5u/L under high-density fermentation while the production process was decreased to 7.5 days. In addition, rLacDx and rLacDe exhibited similar enzymatic characters in oxidizing substrate guaiacol, and were stable at 50℃ and at a pH range from 3 to 10. However, the specific activity of rLacDx (1,761u/mg) for ABTS was higher than that of rLacDe (1,122u/mg), and the apparent K_m value of rLacDx (427 μM) was less than that of rLacDe (604 μM).
【Fund】: 国家自然科学基金(Nos30370045 30470056和30670069);; 安徽省优秀青年基金(No04043048);; 安徽省教育厅自然基金重点(No2006KJ049A);; 国家863高技术研究与发展计划项目(No2007AA09Z421);; 安徽大学211创新团队计划(No02203109)资助~~
【CateGory Index】: Q78
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1 Chen Wanru Li Zhentang Shi Meizhong Chen Huayong Ye Xuanyi.;A Preliminary Study on the Culture of Terraitospheria duthiei[J];JOURNAL OF HANGZHOU UNIVERSITY(NATURE SCIENCE);1989-01
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