High output of a Trametes laccase in Pichia pastoris and characterization of recombinant enzymes / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1055-1059, 2007.
Article
in Zh
| WPRIM
| ID: wpr-276162
Responsible library:
WPRO
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 1.21 x 10(5) u/L and 7.38 x 10(4) u/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 x 10(5) 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 degrees 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 microM) was less than that of rLacDe (604 microM).
Full text:
1
Index:
WPRIM
Main subject:
Pichia
/
Recombinant Proteins
/
Cloning, Molecular
/
Laccase
/
Trametes
/
Fermentation
/
Genetics
/
Isoenzymes
/
Metabolism
Language:
Zh
Journal:
Chinese Journal of Biotechnology
Year:
2007
Type:
Article