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《Journal of Dalian University of Technology》 2002-03
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Expression of gene(phyA Ⅰ) encoding phytase in pichia pastoris and study of properties of phytase

ZHANG Ling\|hua 1 ,AN Li\|jia 1 , YUAN Xiao\|dong 2, GAO Xiao\|rong 1, WANG Yun\|ji 2( 1.Dept.of Bioeng.,School of Chem. Eng.,Dalian Univ.of Technol.,Dalian 116012, China ; 2. Dept.of Food Sci. and Bioeng.,Dalian Inst. of Light Ind., Dalian 116  
Gene phyAⅠ encoding phytase from Aspergillus ficuum As3.324 was amplified by PCR .Sequencing analysis shows that the gene phyA Ⅰis 1 515 bp in length and contains an intron of 111 nucleodides.The putative protein is composed of 467 amino acids and contains 10 potencial N\|glycosylation sites. There is a signal peptide from the 19 residues of the N\|terminus.The expression plasmid vector pPIC9K/ phyAⅡ for tranforming yeast was constructed.The pPIC9K/phyAⅡ was transformed into pichia pastoris GS115 by electroporation and the recombinant yeast GS115/ phyAⅡ was obtained. The phytase activity of the yeast recombinant GS115/ phyAⅡ is 33 times higher than that of A.ficuum As3.324 . The phytase of GS115/phyAⅡ has two activity peaks at pH 2.5 and 4.5, an optimum temperature of 60℃ and its thermostability is also improved significantly in comparison with that of the phytase produced by As3.324.
【Fund】: 辽宁省自然科学基金资助项目 (2 2 2 0 60 )
【CateGory Index】: Q814
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