title: [Gene cloning and expression of serine protease SFP2 from Streptomyces fradiae var. k11].
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.
authors.affiliation: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Li
authors.forename: Peng-Jun
authors.initials: PJ
authors.lastname: Shi
authors.forename: Wang-Zhao
authors.initials: WZ
authors.lastname: Zhang
authors.forename: Xiao-Yu
authors.initials: XY
authors.lastname: Han
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Xu
authors.forename: Hui-Tu
authors.initials: HT
authors.lastname: Zhang
authors.forename: Bin
authors.initials: B
authors.lastname: Yao
authors.forename: Yun-Liu
authors.initials: YL
authors.lastname: Fan
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics
chemical_list: D012697:Serine Endopeptidases