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Influence of Polyols on the Stability and Kinetic Parameters of Invertase from Candida utilis: Correlation with the Conformational Stability and Activity

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Abstract

Invertase (β-d-fructofuranoside fructohydrolase-E.C. 3.2.1.26) is a sucrose hydrolyzing enzyme found in microbial, plant and animal sources. Invertase from Candida utilis is a dimeric glycoprotein composed of two identical monomer subunits with an apparent molecular mass of 150 kDa. We investigated the mechanism of stabilization of invertase with polyols (glycerol, xylitol, and sorbitol). Activity, thermodynamic and kinetic measurements of invertase were performed as a function of polyol concentration and showed that polyols act as very effective stabilizing agents. The result from the solvent-invertase interaction shows preferential exclusion of the polyols from the protein domain leading to preferential hydration of protein. Apparent thermal denaturation temperature of the protein (T m ) rose from 75 °C to a maximum of 85 °C in 30% glycerol. The stabilization has been attributed to the preferential hydration of the enzyme.

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Abbreviations

T m :

Midpoint temperature of thermal unfolding

UV:

Ultraviolet

CD:

Circular dichroism

kDa:

Kilodalton

ξ3 :

Preferential interaction parameter

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Acknowledgement

Gangadhara gratefully acknowledges financial support from the Council of Scientific and Industrial Research (CSIR), New Delhi for providing Senior Research Fellowship.

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Authors and Affiliations

  1. Department of Protein Chemistry and Technology, Central Food Technological Research Institute (A Constituent Laboratory of Council of Scientific and Industrial Research), Mysore, 570 020, India

    Gangadhara, Parigi Ramesh Kumar & Vishweshwaraiah Prakash

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  1. Gangadhara
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  2. Parigi Ramesh Kumar
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  3. Vishweshwaraiah Prakash
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Correspondence to Vishweshwaraiah Prakash.

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Gangadhara, Ramesh Kumar, P. & Prakash, V. Influence of Polyols on the Stability and Kinetic Parameters of Invertase from Candida utilis: Correlation with the Conformational Stability and Activity. Protein J 27, 440–449 (2008). https://doi.org/10.1007/s10930-008-9154-z

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  • DOI: https://doi.org/10.1007/s10930-008-9154-z

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