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

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 degrees 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.

A novel alkaline protease from alkaliphilic Idiomarina sp. C9-1 with potential application for eco-friendly enzymatic dehairing in the leather industry

“…Some of them have been well characterized 6 – 16 . In addition to Bacillus bacterial, alkaline proteases from several gram-negative bacteria such as Pseudomonas 17 , Xanthomonas 18 , and Vibrio 19 have also been found. Moreover, fungi especially genus Aspergillus 20 , 21 , yeast 22 , 23 , and actinomycetes 24 , 25 are also prolific sources for alkaline proteases.…”