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Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)

Title:		[Influence of signal peptide sequences on the expression of heterogeneous proteins in Pichia pastoris]
Author(s):		Xiong AS; Peng RH; Li X; Fan HQ; Yao QH; Guo MJ; Zhang SL;
Address:		"Agro-Biotechnology Research Center of Shanghai Academy of Agricultural Sciences, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Shanghai 201106, China"
Journal Title:		Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)
Year:		2003
Volume:		35
Issue:		2
Page Number:		154 - 160
DOI:
ISSN/ISBN:		0582-9879 (Print) 0582-9879 (Linking)
Abstract:		"Pichia pastoris 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 Saccharomyces 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 of that with original Saccharomyces cerevisiae mating factor alpha 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"
Keywords:		"6-Phytase/metabolism Alcohol Oxidoreductases/genetics Amino Acid Sequence Amino Acids/genetics Base Sequence Electrophoresis, Polyacrylamide Gel Gene Expression Mating Factor Molecular Sequence Data Peptides/genetics Pichia/*genetics/metabolism Plasmids/g;"
Notes:		"MedlineXiong, Ai-Sheng Peng, Ri-He Li, Xian Fan, Hui-Qin Yao, Quan-Hong Guo, Mei-Jin Zhang, Si-Liang chi Research Support, Non-U.S. Gov't China 2003/01/25 Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Feb; 35(2):154-60"