Purification of a protease inhibitor from Dolichos biflorus using immobilized metal affinity chromatography.

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dc.contributor.authorKuhar, Kalika
dc.contributor.authorMittal, Anuradha
dc.contributor.authorKansal, Rekha
dc.contributor.authorGupta, Vijay Kumar
dc.date.accessioned2014-12-26T10:01:53Z
dc.date.available2014-12-26T10:01:53Z
dc.date.issued2014-02
dc.description.abstractPlant protease inhibitors (PIs) are generally small proteins which play key roles in regulation of endogenous proteases and may exhibit antifeedant, antifungal, antitumor and cytokine inducing activities. Dolichos biflorus (horse gram) is an unexploited legume, which is rich in nutrients and also has therapeutic importance. It contains a double-headed PI, which is an anti-nutritional factor. As there is no report available on its simultaneous removal and purification in single step, in this study, a double-headed PI active against both trypsin and chymotrypsin was purified from Dolichos biflorus to ~14-fold with ~84% recovery using an immobilized metal affinity chromatography (IMAC) medium consisting of Zn-alginate beads. The method was single-step, fast, simple, reliable and economical. The purified inhibitor showed a single band on SDS-PAGE corresponding to molecular mass of 16 kDa and was stable over a pH range of 2.0-12.0 and up to a temperature of 100°C for 20 min. The optimum temperature for trypsin and chymotrypsin inhibitor was observed to be 50°C and 37°C, respectively and pH optimum was pH 7.0 and 8.0, respectively. Thus, IMAC using Zn-alginate beads was useful in simultaneous purification and removal of an anti-nutritional factor from horse gram flour in single step. This procedure may also be employed for purification of other plant PIs in one step.en_US
dc.identifier.citationKuhar Kalika, Mittal Anuradha, Kansal Rekha, Gupta Vijay Kumar. Purification of a protease inhibitor from Dolichos biflorus using immobilized metal affinity chromatography. Indian Journal of Biochemistry & Biophysics. 2014 Feb; 51(1): 66-74.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/154237
dc.language.isoenen_US
dc.source.urihttps://nopr.niscair.res.in/handle/123456789/27289en_US
dc.subjectTrypsinen_US
dc.subjectChymotrypsinen_US
dc.subjectDolichos biflorusen_US
dc.subjectImmobilized metal affinity chromatographyen_US
dc.subjectProtease inhibitoren_US
dc.subjectPurificationen_US
dc.subject.meshAlginates --chemistry
dc.subject.meshChromatography, Affinity --methods
dc.subject.meshDolichos --chemistry
dc.subject.meshHydrogen-Ion Concentration
dc.subject.meshMicrospheres
dc.subject.meshMolecular Weight
dc.subject.meshPlant Proteins --chemistry
dc.subject.meshPlant Proteins --isolation & purification
dc.subject.meshProtease Inhibitors --chemistry
dc.subject.meshProtease Inhibitors --isolation & purification
dc.subject.meshProtein Stability
dc.subject.meshTemperature
dc.subject.meshZinc --chemistry
dc.titlePurification of a protease inhibitor from Dolichos biflorus using immobilized metal affinity chromatography.en_US
dc.typeArticleen_US
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