Gene cloning and expression of serine protease SFP2 from Streptomyces fradiae var. k11.
- Author:
Jiang LI
1
;
Peng-Jun SHI
;
Wang-Zhao ZHANG
;
Xiao-Yu HAN
;
Ling-Ling XU
;
Hui-Tu ZHANG
;
Bin YAO
;
Yun-Liu FAN
Author Information
1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Bacillus subtilis;
genetics;
metabolism;
Base Sequence;
Escherichia coli;
genetics;
metabolism;
Molecular Sequence Data;
Serine Endopeptidases;
genetics;
metabolism;
Streptomyces;
enzymology;
genetics
- From:
Chinese Journal of Biotechnology
2005;21(5):782-788
- CountryChina
- Language:Chinese
-
Abstract:
Extracellular serine protease SFP2 from Streptomyces fradiae var. k11 with high feather-degrading activity was purified. The partial amino acid sequences of internal peptide of purified SFP2 were determined, and the partial gene encoding SFP2 was cloned by PCR using the degenerate primers designed according to the amino acid sequences. Complete sfp2 gene was cloned by screening the genomic DNA library of Streptomyces fradiae var. k11. The Open Reading Frame of sfp2 including pre- pro-enzyme is 924bp long (EMBL Accession number: AJ784940). The signal peptide sequence is as long as 114bp, the precursor sequence is 810bp and the mature enzyme is 576bp long, encoding 191 amino acid resides with the putative molecular weight of 19.112kD. In E. coli and Bacillus subtilis, the two sequences encoding SFP2 pro-enzyme and mature enzyme were both expressed successfully. The pro-enzyme expressed had normal biological function and its mature product had normal enzymatic activity.
