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Applied and Environmental Microbiology, May 1999, p. 1915-1918, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae

Yanming Han,1 David B. Wilson,2 and Xin gen Lei1,*

Department of Animal Science1 and Section of Biochemistry, Molecular and Cell Biology,2 Cornell University, Ithaca, New York 14853

Received 21 September 1998/Accepted 10 February 1999

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals and reduces phosphorus pollution of animal waste. Our objectives were to express an Aspergillus niger phytase gene (phyA) in Saccharomyces cerevisiae and to determine the effects of glycosylation on the phytase's activity and thermostability. A 1.4-kb DNA fragment containing the coding region of the phyA gene was inserted into the expression vector pYES2 and was expressed in S. cerevisiae as an active, extracellular phytase. The yield of total extracellular phytase activity was affected by the signal peptide and the medium composition. The expressed phytase had two pH optima (2 to 2.5 and 5 to 5.5) and a temperature optimum between 55 and 60°C, and it cross-reacted with a rabbit polyclonal antibody against the wild-type enzyme. Due to the heavy glycosylation, the expressed phytase had a molecular size of approximately 120 kDa and appeared to be more thermostable than the commercial enzyme. Deglycosylation of the phytase resulted in losses of 9% of its activity and 40% of its thermostability. The recombinant phytase was effective in hydrolyzing phytate phosphorus from corn or soybean meal in vitro. In conclusion, the phyA gene was expressed as an active, extracellular phytase in S. cerevisiae, and its thermostability was affected by glycosylation.

* Corresponding author. Mailing address: 252 Morrison Hall, Department of Animal Science, Cornell University, Ithaca, NY 14853-4801. Phone: (607) 254-4703. Fax: (607) 255-9829. E-mail: XL20{at}cornell.edu.

Applied and Environmental Microbiology, May 1999, p. 1915-1918, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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