Journal: Journal of Pharmacy Research

Article Id: JPRS-BMB-00001220
Title: Effects of Osmolytes on The Structural Stability of Bovine Trypsin: A Brief Review
Category: Biochemistry and Molecular Biology
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    Proteases represent a formidable class of industrial enzymes, accounting for about half of the total sale of the enzymes in the world. Trypsin, a protease that has strong specificity, acts as a digestive enzyme and plays a pivotal role in digestive physiology. Proteins have evolved to function within the cellular milieu, where macromolecule and small molecule solutes are present at high concentrations. It has long been known that osmolytes and other solutes transmute protein stability in vitro while certain osmolytes stabilize proteins in vivo to thermal and chemical denaturation arising in stress conditions. The role of various osmolytes and denaturants in the conformational properties and stability of the enzyme, trypsin has been scarcely investigated. In this study an attempt has been made to probe the effects of osmolytes, Glucose, Glycine, Proline, Sucrose, Trehalose and denaturants Guanidine Hydrochloride and Urea along with their concentration effects on the activity and stability of bovine trypsin. Detailed circular dichroism, steady state and temperature dependent fluorescence spectra, UV-Vis spectra and enzyme assay have been studied to scrutinise osmolyte induced stability and denaturant-induced unfolding properties of trypsin. The specific activity peak was found to be highest in sucrose and lowest in urea and GnHCl. The UV-Vis spectroscopic data showed highest absorbance peak for Proline depicting it as the strongest stabilizer, supported by low absorbance peak in fluorescence spectra. On the other hand, Urea was found to be strong denaturant with its lowest absorbance peak and highest intensity peak in UV-Vis and fluorescence spectra respectively. High osmolyte concentration demonstrated an increase in stability in case of Proline and significant rise in denaturing capacity of urea. CD analysis depicted highest positive peak in Proline, concluding it as the strongest stabilizer followed by Sucrose and Trehalose. Alpha helix and beta sheet content was estimated by K2D2 software which supported the trend in stability concordant with the rest of the data.

    Cite this article as:Shivani Jain, Sarvesh Seechurn, Parth Gupta, Gaurika Garg,Bhavuk Dhamija, N. Latha, and Kameshwar Sharma YVR,Effects of Osmolytes on The Structural Stability of Bovine Trypsin: A Brief Review, Journal of Pharmacy Research 2015,9(8),500-508.

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    Author(s) Name:

    Shivani Jain1, Sarvesh Seechurn1, Parth Gupta2, Gaurika Garg2, Bhavuk Dhamija2, N. Latha2, and Kameshwar Sharma YVR2*

    Affiliation(s) Name:

    1Biological Sciences, Sri Venkateswara College, University of Delhi, Delhi-110021
    2Department of Biochemistry, Sri Venkateswara College, University of Delhi, Delhi-110021

    Received on:12-01-2015; Revised  on: 11-04-2015; Accepted on: 19-08-2015   

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    Author:

    Shivani Jain1, Sarvesh Seechurn1, Parth Gupta2, Gaurika Garg2, Bhavuk Dhamija2, N. Latha2, and Kameshwar Sharma YVR2*

    Title:Effects of Osmolytes on The Structural Stability of Bovine Trypsin: A Brief Review
    Journal:Journal of Pharmacy Research
    Vol(issue):9(8)
    Year:2015
    Page No: (500-508)
  • Experimental Methods Keywords

    Methodology:Protein folding, denaturants, spectroscopy
    Research Materials:Trypsin, protease, osmolytes

Keywords

Trypsin protease osmolytes protein folding denaturants spectroscopy enzyme kinetics

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