[Recombinant batroxobin expressed highly in Pichia pastoris].
Methylotrophic yeast, Pichia pastoris was used to express recombinant batroxobin, and a technology route of producing recombinant protein was finally established. We synthesized batroxobin gene artificially by means of recursive PCR. pPIC9-batroxobin was constructed and transformed into Pichia pastoris GS115 (his4). Recombinant batroxobin was expressed in yeast engineering strain and it was purified from the culture supernatant. 10 mg of recombinant batroxobin was purified from 1 liter fermentation media, it exhibited specific activity of 238 NIH units/mg and had molecular weight of 30.55 kD. The purified recombinant protein converted fibrinogen into fibrin clot in vitro, and shortened bleeding time in vivo. This study laid a foundation of development of hemostatic of recombinant snake venom thrombin-like enzyme.
Li
Zhao-Fa
ZF
Institute of Bioengineering, Academy of Military Medical Science, Beijing 100071, China.
Yu
Xue-Ling
XL
Huang
Jin-Lu
JL
Fang
Hong-Qing
HQ
Chen
Hui-Peng
HP
D000818:Animals; D012103:Batroxobin / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D006470:Hemorrhage / Q000517:prevention & control; D006863:Hydrogen-Ion Concentration; D008297:Male; D051379:Mice; D010843:Pichia / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism* / Q000494:pharmacology; D013997:Time Factors
D011994:Recombinant Proteins; D012103:Batroxobin