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HomeAdvanced Materials ResearchAdvanced Materials Research Vols. 1033-1034Site-Directed Mutagenesis of a Neutral Phytase...

Site-Directed Mutagenesis of a Neutral Phytase from Bacillus amyloliquefaciens: Influencing Activity and Stability

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Abstract:

In order to improve the activity and stability of phytase from Bacillus amyloliquefaciens, site-directed mutagenesis has been performed base on the previous recombinant E.coli BL21 harboring the expression vector of phyC. Mutation residues were chosen based on the sequence alignments and structure analysis of neutral phytsaes from different microorganisms. Site-directed mutagenesis techniques were used to get three mutants (D148E/H149R, Q67E/N68R, and D191E), then the mutants were expressed and purified. Enzymatic characters of different mutants were investigated. The results indicated that the optimum pH of all mutants were 7.0, and the optimum temperature were between 65 °C–70 °C. The maximum specific activity of mutant D148E/H149E was 27.84 U/mg which was 2.19 times than that of the wild-type phytase. The half inactivation temperature of D191E was 4.5 °C higher than that of the wild-type phytase. Fluorescence emission spectra showed that slight differences were among the structures of the mutant phytases. The phytases described here which have increased activity and thermostability may have promosing potential as feed additives in animal diets.

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Advances in Chemical Engineering and Advanced Materials IV

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Periodical:

Advanced Materials Research (Volumes 1033-1034)

Pages:

271-278

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https://doi.org/10.4028/www.scientific.net/AMR.1033-1034.271

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Online since:

October 2014

Authors:

Wei Xu*, Zu Peng Wang, Rong Shao

Keywords:

Enzymatic Character, Neutral Phytase, Site-Directed Mutagenesis

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© 2014 Trans Tech Publications Ltd. All Rights Reserved

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