Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.
 

Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.

Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.

Protein Pept Lett. 2010;17(10):1228-33

Authors: Wang J, Zhang Y, Li S, Liu X, Yan X, Song H, Yu M, Dai L, Mo W

The novel recombinant hirudin, r-RGD-hirudin, inhibits thrombin and platelet aggregation. Here, we reported over-expression of (15)N-labeled r-RGD-hirudin by Pichia pastoris in minimal medium. After extensive optimization, the yield of active r-RGD-hirudin reached ?600 mg/L when the yeast cells were culture in a fermenter. The purified (15)N-labeled r-RGD-hirudin retained full biological activity and was uniformly labeled. Heteronuclear NMR of the (15)N-labeled r-RGD-hirudin was performed for the first time, and all signals in the heteronuclear single quantum coherence (HSQC) spectrum were successfully assigned.

PMID: 20450489 [PubMed - indexed for MEDLINE]



Source: PubMed