TITLE:
Elimination of a disulfide bridge in Aspergillus niger NRRL 3135 Phytase (PhyA) enhances heat tolerance and optimizes its temperature versus activity profile
AUTHORS:
Edward Mullaney, Kandan Sethumadhavan, Stephanie Boone, Xin Gen Lei, Abul H. J. Ullah
KEYWORDS:
Phytase; Disulfide Bridge; Aspergillus niger; Site-Directed Mutagenesis
JOURNAL NAME:
Advances in Biological Chemistry,
Vol.2 No.4,
November
23,
2012
ABSTRACT: In this study, the optimum temperature was lowered while the residual phytase activity after heating to 70℃ was raised in a widely utilized phytase, Aspergillus niger NRRL 3135 PhyA. This was accomplished by site-directed mutagenesis of the cysteines that are involved in the formation of a single disulfide bridge (DB). When compared to wild type (WT), three of the four mutant phytases displayed a lower optimum temperature, 42℃, and up to a four-fold increase in activity after heating. These findings have a potentially broad application to be incorporated along with other desirable features to engineer a phytase with superior physical and chem-ical attributes for animal feed applications.