Chitinase Gene Overexpression of Chaetomium cupreum in Pichia pastoris by Multi-site Mutations
- VernacularTitle:角毛壳菌几丁质酶基因chi58多点突变及在毕赤酵母中的高效表达
- Author:
Yan-Jun WANG
;
Qian YANG
;
- Publication Type:Journal Article
- Keywords:
Chaetomium cupreum, Chitinase, Multi-site mutations, Pichia pastoris, Overexpression
- From:
Microbiology
2008;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
By using gene SOEing PCR, Arg (CGC) in the expression fragment of chitinase gene (chi58) from Chaetomium cupreum was mutated synonymously to Arg (AGA), which is a bias code of Pichia pas-toris yeast. The expression plasmid pPIC9K-chi58A was constructed and transformed into GS115 strain through electroporation. The chitinase activity reached 101.71 U/mL?3.33 U/mL after induced with 120 h, which was the 3 fold of the original strain (31.83 U/mL?4.85 U/mL). Additionally, recombinant yeast showed good genetic stability after 10 cycle. SDS-PAGE analysis suggested that the weight of protein was 58 kD.
