title: Production of horsegram (Dolichos biflorus) Bowman-Birk inhibitor by an intein mediated protein purification system.
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.
authors: Department of Protein Chemistry and Technology, CSIR-Central Food Technological Research Institute, Mysore 570 020, India. kvinod13@gmail.com
authors: Vinod
authors: V
authors: Kumar
authors: Lalitha R
authors: LR
authors: Gowda
mesh_terms: D000818:Animals; D002417:Cattle; D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000737:chemistry; D031294:Dolichos / Q000737:chemistry*; D047668:Inteins; D012639:Seeds / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000737:chemistry / Q000302:isolation & purification*; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D014361:Trypsin Inhibitors; D002918:Chymotrypsin; D014357:Trypsin