Sheng wu gong cheng xue bao = Chinese journal of biotechnology · 2009
[Expression of snake venom thrombin-like enzyme calobin in Pichia pastoris].
Аннотация на английскомСтатьяНазвание и аннотация: Перевод в очереди
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Yuan Shengling, Wang Peng, Tao Haoxia, Zhan Dewen, Wang Yanchun, Wang Lingchun, Liu Chunjie, Zhang Zhaoshan
Аннотация
Thrombin-like enzymes (TLEs) are studied widely because of their therapeutic potential in myocardial infarction and thrombotic diseases. We synthesized the DNA fragment encoding thrombin-like enzyme calobin from Agkistrodon caliginosus (Korean Viper) venom by fusion PCR and expressed it in Pichia pastoris. After induction by 0.5% methanol for 48 h, the expression level of recombinant calobin reached 3.5 g/L in medium. The recombinant calobin was purified by Q-Sepharose Fast Flow ion-exchange chromatography and Sephacryl-S-100 gel filtration chromatography. Purified sample had a molecular weight of 32 kD shown in SDS-PAGE. It hydrolyzed fibrinogen and formed a light white hydrolysis circle in fibrinogen plate. SDS-PAGE analysis showed that recombinant calobin cleaved Aalpha-chain of fibrinogen specifically, and produced an appropriately 40 kD new band. However, we failed to find its fibrin-clot formation activity.