Cloning of Lumbrokinase from earthworm and its expression in Pichia pastoris GS115
We have cloned and expressed a novel earthworm fibrinolytic enzyme (EFE) of Lumbricus rubellus in Pichia pastoris. Its cDNA sequence (GenBank Accession No. DQ202401) revealed a 738 bp region containing an intact ORF that encodes a protein of 245 amino acid residues, containing a signal peptide of 7 amino acid residues and a mature peptide of 238 amino acid residues, designated as EFE F238. Its cDNA shows a high degree of sequence homologies with four other EFE cDNAs registered in GenBank. The gene encoding the native form of F238, with a 5’ non-functional end removed, was obtained by PCR amplification and was sub-cloned into pPIC9, a yeast expression and secretion vector. SDS-PAGE analyses showed that EFE F238 could be secreted into the culture medium. The artificial fibrin plate experiment demonstrated that EFE F238 was active, showing a fibrinolytic activity of about 130 urokinase units/mg protein in basal salts medium.
Ming Li Tao Niu Mingming Zhao Junhuan Zhang Dongjun Kong
Tianjin Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology, Tianjin Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology,
国际会议
成都
英文
1-4
2010-06-18（万方平台首次上网日期，不代表论文的发表时间）