Over-expression and Purification of Formate Dehydrogenase form Pichia pastoris in Escherichia coli / 微生物学通报
Microbiology
;
(12)1992.
Article
in Chinese
| WPRIM
| ID: wpr-685313
ABSTRACT
Formate dehydrogenase(FDH)coding gene was amplified from genomic DNA of Pichia pastoris by polymerase chain reaction, and the codon TAG(bases 649-651)was mutated to GAG using site-directed mutagenesis.The recombinant plasmid pET-FDH was con- structed by inserting the mutated DNA fragment into expression vector pET-22b(+),and transformed into E.coli BL21(DE3).FDH was expressed as a form of soluble prutein fused with 6?His tag at high level through IPTG induction.The amount of FDH was up to about 30% of the total cell protein.The cells-free crude extract was purified by one affinity chromatographic step,and resulting enzyme preparation revealed a specific activity of 6.45 U/mg.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Microbiology
Year:
1992
Type:
Article
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