Heterologous Expression and Functional Characterization of a Truncated Marine Alginate Lyase
Qianqian Shao1, Chaoying Yao1, Xingbin Wang1
1State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
Abstract:
Alginate lyase is widely used in the preparation of alginate oligosaccharides, medicine production, energy conversion, and so on. In this study, we designed truncated mutants of a marine-derived alginate lyase Algl and identified a highly active mutant CD317 with less degradation when expressed in a yeast host. The enzyme activity of the secretory CD317 was 1.4-fold higher than that of the parent Algl. It degraded both polyM and polyG but had a stronger preference for polyM. The optimal temperature of Algl and CD317 was 40 °C and 35 °C respectively, for which CD317 showed better temperature tolerance. Additionally, Ca2+ highly improved the enzyme activity. Fermentation conditions for CD317 production were optimized as a culture time of 144 h, an inoculum of an OD600nm of 0.5, and an inducer concentration of 2% (v/v), respectively. Bioreactor fermentation allowed the highest production of 19,500 U/mL, which was 4.6-fold higher than that in a well-plate. These results indicated that the truncated CD317 showed good potential for industrial use.
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