Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K

✍ Scribed by Raja Noor Zaliha Raja Abd. Rahman; Lee Poh Geok; Chee Fah Wong; Mahiran Basri; Abu Bakar Salleh

• Publisher: John Wiley and Sons
• Year: 2010
• Tongue: English
• Weight: 207 KB
• Volume: 50
• Category: Article
• ISSN: 0233-111X
• DOI: 10.1002/jobm.200900133

✦ Synopsis

Abstract

A gene encoding an organic solvent‐stable protease was amplified from Pseudomonas aeruginosa strain K by polymerase chain reaction using consensus primers based on multiple sequence alignment of alkaline and metalloprotease genes from Pseudomonas species. The gene, which consisted of 1440 bp nucleotides and deduced 479 amino acid residues, was successfully expressed in pGEX‐4T‐1 expression system in the presence of 1.0 mM IPTG, after an incubation of 6 h at 37 °C. Under these conditions, the recombinant strain K protease was, subsequently, released into the periplasm of E. coli BL21 (DE3) with an optimum proteolytic activity detected at 1.0112 U/ml. To date, this is the first reported expression of alkaline protease (aprA) with such remarkable property in Escherichia coli. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)