A novel extracellular protease from Pseudomonas aeruginosa MCM B-327: enzyme production and its partial characterization.
The focus of this study was on production, purification and characterization of dehairing protease from Pseudomonas aeruginosa MCM B-327, isolated from vermicompost pit soil. Optimum protease activity, 395 U mL(-1), was observed in the medium containing soybean meal and tryptone, at pH 7 and 30 °C. The crude enzyme exhibited dehairing activity. As compared to chemical method, enzymatic method of dehairing showed reduction in COD, TDS and TSS by 34.28%, 37.32% and 51.58%, respectively. Zymogram of crude enzyme on native-PAGE presented two bands with protease activity of molecular weights of 56 and 67 kDa. Both proteases showed dehairing activity. Out of these, 56kDa protease (PA02) was purified 3.05-folds with 2.71% recovery. The enzyme was active in pH range 7-9 and temperature 20-50 °C with optimum pH of 8 and temperature 35°C. Moreover, the enzyme activity of PA02 protease was not strongly inhibited by specific inhibitor showing the novel nature of enzyme compared to serine, cysteine, aspartyl and metalloproteases. Kinetic studies indicated that substrate specificity of PA02 protease was towards various natural and synthetic proteolytic substrates but inactive against collagen and keratin. These findings suggest protease secreted by P. aeruginosa MCM B-327 may have application in dehairing for environment-friendly leather processing.
Zambare
Vasudeo
V
Microbial Sciences Division, Agharkar Research Institute, Pune 411004, India.
Nilegaonkar
Smita
S
Kanekar
Pradnya
P
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002020:Buffaloes; D010450:Endopeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D006197:Hair / Q000378:metabolism; D007218:Industrial Microbiology; D007477:Ions / Q000737:chemistry; D008670:Metals / Q000737:chemistry; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013379:Substrate Specificity; D013633:Tanning / Q000379:methods; D013696:Temperature
D001426:Bacterial Proteins; D007477:Ions; D008670:Metals; D010450:Endopeptidases