Production optimization of extracellular amidase enzyme by newly isolated Pseudomonas putida AP-2 from agricultural soil.

Amidases are ubiquitous enzymes that have received increased attention due to their wide range of biotechnological applications, especially in industries for the synthesis of wide variety of carboxylic and hydroxamic acids, which find applications in pharmaceuticals, agrochemicals and waste water treatments. In the present study, 40 bacterial isolates were screened for extracellular amidase-producing capability, and on the basis of color development, 5 isolates were selected for amidase production in broth media. Based on enzyme production, one of the most potent isolates identified as Pseudomonas putida AP-2 was selected for further study. The effects of media composition and various fermentation conditions for optimization of amidase production were studied. The maximum extracellular amidase production was obtained at 30 °C and pH 8.0 after 36 h of incubation in shaking condition. Among the substrate, acetamide was the best; however, P. putida AP-2 also utilized acrylamide which is a known carcinogen. Regarding carbon sources, glucose was the best, while peptone was found the best nitrogen source. The isolated bacterium, P. putida AP-2, is also tolerant to number of heavy metals at higher levels so this may also be applied for field application in contaminated soil. [ABSTRACT FROM AUTHOR]