Objective: The marine catalase YS0810CAT gene was cloned and overexpressed from Acinetobacter sp. YS0810, and high stability of the recombinant enzyme was validated, which made it important for potential applications in the elimination of hydrogen peroxide from industrial process-generated streams.
Methods: The gene was cloned by PCR and overexpressed in Escherichia coli. Evolutionary analyses of this enzyme were conducted with the MEGA software. Anion exchange was applied to purify the recombinant enzyme. The effects of pH and temperature on the activity and stability of YS0810CAT were measured.
Results: The gene consists of 1,518 bp and belongs to Clade 3 of monofunctional catalases. The maximum protein production was obtained with 0.8 mM IPTG, a post-induction temperature of 37°C, and a post-induction time of 8 h. The recombinant protein was most active at 60°C and pH 11.
Conclusion: The effects of pH and temperature on the activity and stability of the wild type and recombinant YS0810CAT are similar. The protocol for the preparation of recombinant YS0810CAT could aid enzyme crystallization; moreover, improvement in its properties may be possible through protein engineering.
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