Production of horsegram (Dolichos biflorus) Bowman-Birk inhibitor by an intein mediated protein purification system

Protein Expression and Purification
Vinod Kumar, L R Gowda

Abstract

The seeds of the legume horsegram (Dolichos biflorus), a protein rich pulse (bean), contain multiple forms of Bowman-Birk inhibitors (protease inhibitors). The major inhibitor HGI-III contains seven interweaving disulfides and is extremely stable to high temperatures. A soluble HGI-III (rHGI) with the native N-terminus was produced using a pTWIN IMPACT™ purification system. Yield of rHGI was improved by introducing a trypsin sepharose affinity chromatography step resulting in ∼670 fold purification. The biochemical characteristics of rHGI point to its close similarity to seed HGI-III not only in its structure but also in its inhibitory characteristics toward bovine trypsin and chymotrypsin. The expression and purification strategy presented here promises to produce BBIs in their natural form for pharmacological and therapeutic use.

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Citations

Mar 25, 2014·Biotechnology Letters·Dongli Guan, Zhilei Chen
Apr 22, 2015·Journal of Food Science and Technology·Saroj Kumar Prasad, Manoj Kumar Singh
Jan 13, 2015·Journal of Industrial Microbiology & Biotechnology·Qing HeTian-jiao Li
Dec 21, 2014·Archives of Biochemistry and Biophysics·Vinod KumarLalitha R Gowda

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