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

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 p...

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Main Authors: Han, Yanming, Wilson, David B., Lei, Xin gen
Formato: Artigo
Idioma:en
Publicado em: American Society for Microbiology 1999
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Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC91276/
https://ncbi.nlm.nih.gov/pubmed/10223979
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spelling pubmed-912762002-09-12 Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae Han, Yanming Wilson, David B. Lei, Xin gen Appl Environ Microbiol Genetics and Molecular Biology 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. American Society for Microbiology 1999-05 /pmc/articles/PMC91276/ /pubmed/10223979 Text en Copyright © 1999, American Society for Microbiology
institution US National Library of Medicine
collection PubMed Central
language en
format Article
topic Genetics and Molecular Biology
spellingShingle Genetics and Molecular Biology
Han, Yanming
Wilson, David B.
Lei, Xin gen
Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae
description 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.
author Han, Yanming
Wilson, David B.
Lei, Xin gen
author_facet Han, Yanming
Wilson, David B.
Lei, Xin gen
author_sort Han, Yanming
title Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae
title_short Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae
title_full Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae
title_fullStr Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae
title_full_unstemmed Expression of an Aspergillus niger Phytase Gene (phyA) in Saccharomyces cerevisiae
title_sort expression of an aspergillus niger phytase gene (phya) in saccharomyces cerevisiae
publisher American Society for Microbiology
publisher_facet American Society for Microbiology
publishDate 1999
url https://ncbi.nlm.nih.gov/pmc/articles/PMC91276/
https://ncbi.nlm.nih.gov/pubmed/10223979
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