title: [Expression of the thermostable carboxypeptidase Taq gene in Pichia pastoris GS115].
abstract: To express recombinant carboxypeptidase from Thermus aquaticus (Cpase Taq) in Pichia pastosis, the open reading frame coding thermostable Cpase Taq was optimized based on the preference of P. pastoris codon usage and synthesized in vitro. The novel gene was cloned into P. pastoris expression vector pHBM905A and the sequence coding 6xHis tag was fused with the ORF of Cpase Taq gene. The recombinant plasmid was named pHBM905A-Cpase Taq and transformed into P. pastoris GS 115. Transformants were induced with 1% methanol for 72 h until the enzyme yield reached 0.1 mg/ml. The enzyme was purified and its enzymatic properties were analyzed. The results showed that the specific enzyme activity reached maximum at 75 °C and pH 7.5, which was about 80 U/mg. It was the first report about the secretory expression of Cpase Taq in P. pastoris GS115. Because of its large-scale preparation, this enzyme may be applied in industrial hydrolysis of peptides into amino acids in the future.
authors.forename: Xianhong
authors.initials: X
authors.lastname: Yu
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Wang
authors.forename: Xing
authors.initials: X
authors.lastname: Zhong
authors.forename: Wei
authors.initials: W
authors.lastname: Tang
authors.forename: Chao
authors.initials: C
authors.lastname: Zhai
authors.forename: Wanping
authors.initials: W
authors.lastname: Chen
authors.forename: Lixin
authors.initials: L
authors.lastname: Ma
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics*; D002268:Carboxypeptidases / Q000096:biosynthesis* / Q000235:genetics*; D003001:Cloning, Molecular; D003062:Codon; D006868:Hydrolysis; D016366:Open Reading Frames; D010843:Pichia / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013824:Thermus / Q000201:enzymology
chemical_list: D001426:Bacterial Proteins; D003062:Codon; D011994:Recombinant Proteins; D002268:Carboxypeptidases; C089478:Thermus aquaticus carboxypeptidase