A collagenase from Pseudomonas sp. was purified upto 13-fold with a yield of 3.6% using hydrophobic interaction chromatography. Molecular weight of 34kDa collagenase was identified using sodium dodecyl sulfate-polyacrylamide gel electrophoresis method. Gelatin and collagen substrates were used in the characterization of purified collagenase. The optimum temperature and pH were 45 °C, pH 7.5 for gelatin and 50 °C, pH 8.5 for collagen respectively. The collagenase activity was highly inhibited by

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... Hg2+, Pb2+, EDTA, DTT at concentration of 1mM and significant negative effect of Fe2+, Cu2+ and Co2+ was observed at similar concentration. However, Ca2+ (1mM) and PEG (1mM) increased the enzyme activity. The Pseudomonas based collagenase have the ability to hydrolyse protein substrates. The K m and V max values for the purified collagenase were found to be 1.65mg/mL and 2.12U/mL with collagen substrate while for gelatin substrate values were 6.60mg/mL and 6.58U/mL, respectively. (Figure 1) Figure 1 : Graphical abstract. Graphical Abstract Highlights A. Screening of soil/sewage samples for the hyper producer strain with maximum collagenase activity B. Strain was identified as Pseudomonas sp. C. Determination of protein content and collagenase activity D. Production, purification and characterization of collagenase enzyme from Pseudomonas sp.