title: Enhanced production and organic solvent stability of a protease from Brevibacillus laterosporus strain PAP04.
abstract: A bacterial strain (PAP04) isolated from cattle farm soil was shown to produce an extracellular, solvent-stable protease. Sequence analysis using 16S rRNA showed that this strain was highly homologous (99%) to Brevibacillus laterosporus. Growth conditions that optimize protease production in this strain were determined as maltose (carbon source), skim milk (nitrogen source), pH 7.0, 40°C temperature, and 48 h incubation. Overall, conditions were optimized to yield a 5.91-fold higher production of protease compared to standard conditions. Furthermore, the stability of the enzyme in organic solvents was assessed by incubation for 2 weeks in solutions containing 50% concentration of various organic solvents. The enzyme retained activity in all tested solvents except ethanol; however, the protease activity was stimulated in benzene (74%) followed by acetone (63%) and chloroform (54.8%). In addition, the plate assay and zymography results also confirmed the stability of the PAP04 protease in various organic solvents. The organic solvent stability of this protease at high (50%) concentrations of solvents makes it an alternative catalyst for peptide synthesis in non-aqueous media.
authors.affiliation: Department of Biological Engineering, Inha University, Incheon, South Korea.
authors.forename: P
authors.initials: P
authors.lastname: Anbu
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry; D058675:Brevibacillus / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D009930:Organic Chemicals / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000096:biosynthesis* / Q000737:chemistry; D012988:Soil Microbiology; D012997:Solvents; D013696:Temperature; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D009930:Organic Chemicals; D012997:Solvents; D010447:Peptide Hydrolases