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Please use this identifier to cite or link to this item: http://apo.ansto.gov.au/dspace/handle/10238/1693

Title: Novel approach for enhancing the catalytic efficiency of a protease at low temperature: reduction in substrate inhibition by chemical modification.
Authors: Siddiqui, KS
Parkin, DM
Curmi, PMG
De Francisci, D
Poljak, A
Barrow, KD
Noble, MH
Trewhella, J
Cavicchioli, R
Keywords: Small Angle Scattering
Enzymes
Crystal Structure
Escherichia Coli
Amylase
Catalytic Effects
Issue Date: 1-Jul-2009
Publisher: Wiley-Blackwell
Citation: Siddiqui, K. S., Parkin, D. M., Curmi, P. M. G., De Francisci, D., Poljak, A., Barrow, K., et al. (2009). Novel approach for enhancing the catalytic efficiency of a protease at low temperature: reduction in substrate inhibition by chemical modification. Biotechnology and Bioengineering, 103(4), 676-686.
Abstract: The alkaline protease, savinase was chemically modified to enhance the productivity of the enzyme at low temperatures on a complex polymeric protein (azocasein) substrate. At 5 and 15 degrees C, savinase modified with ficol or dextran hydrolyzed fivefold more azocasein than the unmodified savinase. Kinetic studies showed that the catalytic improvements are associated with changes in uncompetitive substrate inhibition with K-i values of modified savinases sixfold higher than the unmodified savinase. Modeling of small-angle scattering data indicates that two substrate molecules bind on opposing sides of the enzyme. The combined kinetic and structural data indicate that the polysaccharide modifier sterically blocks the allosteric site and reduces substrate inhibition. In contrast to the properties of cold-active enzymes that generally manifest as low activation enthalpy and high flexibility, this study shows that increased activity and productivity at low temperature can be achieved by reducing uncompetitive substrate inhibition, and that this can be achieved using chemical modification with an enzyme in a commercial enzyme-formulation. Biotechnol. Bioeng. 2009;103: 676-686. © 2009, Wiley-Blackwell. The definitive version is available at www3.interscience.wiley.com
URI: http://dx.doi.org/10.1002/bit.22300
http://apo.ansto.gov.au/dspace/handle/10238/1693
ISSN: 0006-3592
Appears in Collections:Journal Articles

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