Abstract
Amylases are the starch degrading enzymes with applications ranging from food, fermentation, textiles to paper industries. In this study, a soil bacterium from the Chittagong industrial area was screened primarily for amylase activity using starch agar media. An attempt has been made to isolate and identify this amylolytic bacterium, optimization of its amylase production and characterization of the crude enzyme. The isolate was identified as Pseudomonas aeruginosa based on microscopic and biochemical tests. Beside this a speciesspecific gene ecfX (146 bp) for P. aeruginosa was amplified by PCR to confirm the identification of the isolate. A broad range of parameters for the optimization of enzyme production based on selection of best carbon and nitrogen sources, optimum pH, temperature, incubation period and effect of additional metal ions on culture media were investigated. Best enzyme activity (343.60 U/mL) was found in the presence of glucose as carbon source with yeast extract as nitrogen source at temperature 37°C after 24h at pH 9 in addition of Mg++ in submerged fermentation broth culture. The Mg++, SDS and Ca++ increased the amylase activity whereas EDTA and Zn++ were found as the inhibitory agents for amylase.
 Jahangirnagar University J. Biol. Sci. 7(2): 99-114, 2018 (December)
Highlights
Starch is a polymer of glucose linked to one another through the glycosidic bond
Amylopectin consists of short α-1, 4 glycosydic bonds linked to linear chains of 10–60 glucose units and α-1, 6 glycosydic bonds linked to side chains with 15–45 glucose units (Muralikrishna & Nirmala, 2005, Sorensen et al, 2004; Tester et al, 2004)
Screening for the amylase production: The bacteria isolated from Industrial soil of Chittagong were screened for amylase production on starch agar medium
Summary
Starch is a polymer of glucose linked to one another through the glycosidic bond. Two types of glucose polymers are present in starch: amylose and amylopectin. Amylases are widely distributed in microbial, plant and animal kingdom They degrade starch and related polymers to yield products characteristic of individual amylolytic enzymes (Aiyer, 2005). Specificity and sensitivity analyses showed the ecfX gene based PCR screening to be highly reliable, giving PCR products of the expected size for all P. aeruginosa strains tested and not amplifying DNA from any of the other Pseudomonas species tested (Lavenir et al, 2007). The present study was mainly focused on the production of amylase from soil isolated Pseudomonas aeruginosa by optimizing various parameters such as carbon and nitrogen sources, pH, temperature, incubation time and metal ions. The identification of Pseudomonas aeruginosa was performed using different morphological and biochemical characterization, which was followed by ecfX gene specific PCR based identification at species level of the organism
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