Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2078867 | Chinese Journal of Biotechnology | 2007 | 5 Pages |
Abstract
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 1.21Ã105 u/L and 7.38Ã104 u/L, respectively, as determined with 2,2â²-azinobis (3-ethylbenzothia-zoline- 6-sulfonic acid) (ABTS) as substrate. The yield of rLacDx was further increased to 2.39Ã105 U/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°C and at a pH range from 3 to 10. However, the specific activity of rLacDx (1761 u/mg) for ABTS was higher than that of rLacDe (1122 u/mg), and the apparent Km value of rLacDx (427 μmol/L) was less than that of rLacDe (604 μmol/L).
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Authors
ZHOU Hong-Min, HONG Yu-Zhi, XIAO Ya-Zhong, CUI Teng-Jiao, WANG Xiao-Tang, PU Chun-Lei,