Overproduction of 15N-Labeled r-RGD-Hirudin in Pichia pastoris for NMR Studies

Title: Overproduction of 15N-Labeled r-RGD-Hirudin in Pichia pastoris for NMR Studies

Volume: 17 Issue: 10

Author(s): Jue Wang, Yanling Zhang, Shiyun Li, Xingang Liu, Xiaomin Yan, Houyan Song, Min Yu, Linsen Dai and Wei Mo

Affiliation:

Keywords: 15N-Labeling, NMR, Pichia pastoris, r-RGD-Hirudin

Abstract: The novel recombinant hirudin, r-RGD-hirudin, inhibits thrombin and platelet aggregation. Here, we reported over-expression of 15N-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 cultured in a fermenter. The purified 15N-labeled r-RGD-hirudin retained full biological activity and was uniformly labeled. Heteronuclear NMR of the 15N-labeled r-RGD-hirudin was performed for the first time, and all signals in the heteronuclear single quantum coherence (HSQC) spectrum were successfully assigned.

Cite this article as:

Wang Jue, Zhang Yanling, Li Shiyun, Liu Xingang, Yan Xiaomin, Song Houyan, Yu Min, Dai Linsen and Mo Wei, Overproduction of 15N-Labeled r-RGD-Hirudin in Pichia pastoris for NMR Studies, Protein & Peptide Letters 2010; 17 (10) . https://dx.doi.org/10.2174/092986610792231465