Improved production of polygalacturonate lyase by combining a pH and online methanol control strategy in a two-stage induction phase with a shift in the transition phase |
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Authors: | Muhammad Salman Qureshi Dongxu Zhang Guocheng Du Jian Chen |
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Institution: | (1) School of Biotechnology, Jiangnan University, 214122 Wuxi, China;(2) Key Laboratory of Industrial Biotechnology (Ministry of Education), School of Biotechnology, Jiangnan University, Wuxi, China |
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Abstract: | Polygalacturonate lyase is a kind of enzyme that is abundantly used in the textile industry for cotton scouring. Previously,
we reconstructed the polygalacturonate lyase gene in Pichia pastoris for the expression of this enzyme. To enhance the production of polygalacturonate lyase (PGL), a combined strategy was formulated
by combining online methanol control and two-stage pH control strategies. For the two-stage pH control strategy during the
growth phase, the pH was controlled at 5.5, and in the induction phase different pH levels were investigated for the optimum
enzyme production. During the online methanol control strategy, the different levels of methanol (v/v) were investigated for
the best enzyme production at pH 5.5. These two strategies were combined together for enhanced PGL productivity, and the induction
phase was divided into two stages in which methanol concentrations were maintained at different levels online. The transition
phase was introduced during the induction phase instead of introducing it after the growth phase. The two-stage combination
strategy was formulated on the bases of methanol consumption of cells, optimal pH, cell viability and the production of polygalacturonate
lyase by P. pastoris. By using this strategy, the production was doubled compared with common conditions, and the highest polygalacturonate lyase
activity reached 1,631 U/ml. This strategy proved to be very useful for the enhancement of polygalacturonate lyase production
by achieving higher cell viability, alcohol oxidase activity and phosphate-related compounds of the cells during the induction
phase. |
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Keywords: | |
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