Factorial Design Based Bench-scale Production of Collagenase by Pseudomonas sp. Found in Protein Waste of Himalayan Region

The current study was focused on factorial design based bench-scale production of collagenase by Pseudomonas sp. Chemical and fermentation conditions including medium components (carbon, nitrogen, and growth supplements) were optimized. The medium containing sucrose, tryptone and gelatin substrate was found to enhance the production of collagenase. The physical parameters (agitation speed and aeration rate) were also optimized. Moreover, the interactive effect of optimized physicochemical parameters using two levels of six factors (26) of factorial design was studied for the maximum collagenase production. Among 64 combinations, the 57th combination was shown maximum 1.43 U/mL collagenase activity. The bench-scale production of collagenase was achieved in a 6 L working volume laboratory fermenter. The bench-scale fermenter produced 2.3-fold enhanced collagenase activity at reduced cultivation time (14th h) in comparison to the shake flask (24th h). The lead combination can be used for the large scale collagenase production in industrial fermenters.