Isolation, Identification and Optimization of Culture Conditions of Bacillus sp. Strain PM1 for Alkalo-thermostable Amylase Production.
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Date
2014-04
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Abstract
Aims: To isolate and optimize the culture conditions for thermo stable and alkaline amylase
production from bacteria.
Study Design: Optimization of different physiological and nutritional parameters for
amylase production and kinetic studies of amylase.
Place and Duration of Study: Soil Samples: Herbal garden of Amity University Haryana,
Gurgaon (Manesar), India, between April 2012 and September 2012.
Methodology: Amylolytic isolates were selected by flooding the nutrient agar plates
containing 2% starch with Lugol solution. Isolates were selected on the basis of higher ratio
of clear zone to colony size and grown in nutrient broth containing 2% starch. The level of
amylase was detected in the culture filtrate. The selected isolate showing maximum
amylase production was identified on the basis of 16S rDNA amplification.
Results: An Alkalo-thermostable amylase producing bacterial isolate from soil was
identified as Bacillus sp. strain PM1 on the basis of 16S rRNA. It yielded 3.5 U/ml of
amylase in medium containing (%) starch 2.0, beef extract 0.5, NaCl 0.5 at 50ºC, pH 7.0 at
180 rpm after 72 h. The optimum pH and temperature for amylase activity was 8.0 and
50°C, respectively. The enzyme exhibited 67% activity after 60 minute incubation at 50ºC. At pH 8.0, the enzyme retained 78% activity after 4 h.
Conclusion: The properties of the isolated enzyme are adequate for its use in starch
processing and baking industry.
Description
Keywords
Amylase, Alkalo-thermostable, Bacillus sp., optimization, starch
Citation
Sharma Kinshuk, Bhutty Sarita, Khurana S M Paul, Kohli Urvashi Kumar. Isolation, Identification and Optimization of Culture Conditions of Bacillus sp. Strain PM1 for Alkalo-thermostable Amylase Production. British Microbiology Research Journal. 2014 Apr; 4(4): 369-380.