Cloning and sequencing of Serratia protease gene

• Published in: Nucleic acids research Vol. 14; no. 14; pp. 5843 - 5855
• Main Authors: Nakahama, Kazuo, Yoshimura, Koji, Marumoto, Ryuji, Kikuchi, Masakazu, Lee, In Sook, Hase, Toshiharu, Matsubara, Hiroshi
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 25.07.1986
• Subjects: Amino Acid Sequence; Bacillus subtilis; Base Sequence; Biological and medical sciences; cloning; Cloning, Molecular; Codon; DNA Restriction Enzymes; Endopeptidases - genetics; Escherichia coli; Fundamental and applied biological sciences. Psychology; Genes; Genes, Bacterial; Genes. Genome; Metalloendopeptidases; Molecular and cellular biology; Molecular genetics; nucleotide sequence; Plasmids; proteinase; Serratia; Serratia - enzymology; Serratia - genetics; Serratia marcescens; Bacillus subtilis; Escherichia coli; Proteinase; Primary structure; Bacillaceae; Bacillales; Gene; Serratia marcescens; Bacteria; Molecular cloning; Escherichieae; Klebsielleae; Enterobacteriaceae
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/14.14.5843

The gene encoding an extracellular metalloproteinase from Serratia sp. E-15 has been cloned, and its complete nucleotide sequence determined. The amino acid sequence deduced from the nucleotide sequence reveals that the mature protein of the Serratia protease consists of 470 amino acids with a molecular weight of 50,632. The G+C content of the coding region for the mature protein is 58%; this high G+C content is due to a marked preference for G+C bases at the third position of the codons. The gene codes for a short pro-peptide preceding the mature protein. The Serratia protease gene was expressed in Escherichia coli and Serratia marcescens; the former produced the Serratia protease in the cells and the latter in the culture medium. Three zinc ligands and an active site of the Serratia protease were predicted by comparing the structure of the enzyme with those of thermolysin and Bacillus subtilis neutral protease.