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Characterization of Processive Endoglucanase Stability in Vitro as an Exogenous Fibre Biocatalyst in Pig Nutrition

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Title: Characterization of Processive Endoglucanase Stability in Vitro as an Exogenous Fibre Biocatalyst in Pig Nutrition
Author: Cheng, Laurence
Department: Department of Animal Biosciences
Program: Animal and Poultry Science
Advisor: Fan, Ming
Abstract: Poor efficiency of dietary fibre utilization limits pork production profit margins and environmental sustainability. This thesis research was to investigate stability of two processive endoglucanases, referred to as GH5-tCel5A1 and GH5-p4818Cel5_2A and overexpressed in the ClearColi® BL21 (DE3) cell. Three-dimensional models predicted presence of cysteine residues on the cellulases’ catalytic sites; and results of in vitro time course experiments further shown these cellulases were susceptible to auto-oxidation by air-borne O2 and were unstable. In vitro cellulases’ stability was also examined under the mimicked porcine gastric and the small intestinal conditions. Eadie-Hofstee inhibition kinetic analyses shown that both cellulases lost significant levels of their initial enzyme activities within 2 – 3 h of incubations and were thus not stable under the porcine gastrointestinal environmental conditions. It is concluded that enzyme protein engineering and alternatively post-fermentation enzyme processing need to be further pursued to enable both cellulases as potentially efficacious exogenous fibre enzymes.
URI: https://hdl.handle.net/10214/26527
Date: 2021
Rights: Attribution-NonCommercial-NoDerivatives 4.0 International


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Attribution-NonCommercial-NoDerivatives 4.0 International Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivatives 4.0 International