Author
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Mertens, Jeffrey |
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Submitted to: Meeting Abstract
Publication Type: Abstract Only Publication Acceptance Date: 8/11/2013 Publication Date: 8/15/2013 Citation: Mertens, J.A. 2013. Kinetic characteristics of polygalacturonase enzymes hydrolyzing galacturonic acid oligomers using isothermal titration calorimetry [abstract]. Society of Industrial Microbiology. p. 90. Interpretive Summary: Technical Abstract: Polygalacturonase enzymes hydrolyze the polygalacturonic acid chains found in pectin. Interest in polygalacturonase enzymes continues as they are useful in a number of industrial processes and conversely, detrimental, as they are involved in maceration of economically important crops. While a good deal of biochemical information is available for polygalacturonase enzymes hydrolyzing polygalacturonic acids, little biochemical information exists on the hydrolysis of galacturonic acid oligomers; largely due to the lack of a convenient assay system. In the work presented here, the kinetic characteristics of polygalacturonases hydrolyzing galacturonic acid oligomers are determined using isothermal titration calorimetry. Hydrolysis of galacturonic acid oligomers is an endothermic process and essentially enthalpically silent with delta-Happ values of 2 kcal/mol and less than 1 kcal/mol for the exo- and endo-enzymes, respectively. Kinetic parameters of hydrolysis for two enzymes characterized follow the trend seen with many glycoside hydrolases in that as oligomer length increases, the catalytic rate and the affinity of the enzyme for the substrate increases. One of the exo-acting polygalacturonase enzymes characterized does not follow this general pattern. The exo-acting polygalacturonase enzymes are subject to product inhibition with determined Ki values significantly higher than Km. This work provides additional insight into biochemistry of this enzyme class and provides a means for comparison of polygalacturonase enzymes isolated from different sources. |
