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

Vinod, Kumar and Lalitha, R. Gowda (2013) Production of horsegram (Dolichos biflorus) Bowman-Birk inhibitor by an intein mediated protein purification system. Protein Expression and Purification, 89. pp. 16-24.

PDF Protein Expression and Purification, Volume 89, Issue 1, May 2013, Pages 16-24.pdf - Published Version Restricted to Registered users only Download (990kB)

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.

Item Type:	Article
Uncontrolled Keywords:	Cloning Expression pTWIN1 Trypsin sepharose Chitin beads
Subjects:	500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 16 Enzyme Chemistry 600 Technology > 08 Food technology > 22 Legumes-Pulses
Divisions:	Protein Chemistry and Technology
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	26 Mar 2013 04:23
Last Modified:	26 Mar 2013 04:23
URI:	http://ir.cftri.res.in/id/eprint/11160

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