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Influence of signal peptide sequences on the expression of heterogeneous proteins in Pichia pastoris

文献类型： 外文期刊

作者： Xiong, AS 1 ; Peng, RH 2 ; Li, X 2 ; Fan, HQ 2 ; Yao, QH 2 ; Guo, MJ 2 ; Zhang, SL 2 ;

作者机构： 1.Shanghai Acad Agr Sci, Agrobiotechnol Res Ctr, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China

2.Shanghai Acad Agr Sci, Agrobiotechnol Res Ctr, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China; E China Univ Sci & Technol, Dept Bioengn, Shanghai 200237, Peoples R China

关键词： Pichia pastoris;synthetic signal sequence;expression vector;high expression

期刊名称：ACTA BIOCHIMICA ET BIOPHYSICA SINICA （ 影响因子：3.848； 五年影响因子：3.67 ）

ISSN： 0582-9879

年卷期： 2003 年 35 卷 2 期

页码：

收录情况： SCI

摘要： Pichia pastor's has been developed to be a very efficient expression host for the heterogeneous proteins since its alcohol promoter was isolated and cloned, and its transformation technique was established. For further improving the secretion expression of heterogeneous proteins, in this research, the signal sequences were studied. At first, the Saccharorayces cerevisiae mating factor alpha prepro-leader sequence was synthesized using successive PCR and designated as MF4I. Then, ten different signal sequences were constructed by adding the N-terminal residues of Pichia pastoris Aox1 protein to the N-terminal of the MF4I. These ten signal sequences were used for directing phytase gene secretion in Pichia pastoris, the secretion of phytase were increased in Pichia pastoris strains containing new signal sequence. Among these strains, the phytase secretion was highest in strain contain signal sequence added with A, I, P three Aox1 N-terminal residues; the phytase secretion of Pichia pastoris was 90 mg/L in flake. The secretion was six-fold than that with original Saccharomyces cerevisiae mating factor a prepro-leader sequence. In addition, insert of ten residues E E A E A E A E P K can further increase the phytase secretion by 35%, the secretion reach 120 mg/L.

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