ISSN: 0973-7510

E-ISSN: 2581-690X

Priti Binita Lakra , Sukantibala Mohapatra and Pratima Ray
1Orissa University of Agriculture and Technology, Center for Post Graduate Studies, Department of Microbiology, Bhubaneswar – 751003, Orissa, India.
J Pure Appl Microbiol. 2014;8(4):3147-3153
© The Author(s). 2014
Received: 11/01/2014 | Accepted: 18/03/2014 | Published: 31/08/2014
Abstract

Over millions of years, mangroves have evolved both morphologically and physiologically to adapt to swampy and saline environments. Similar adaptive characteristics in the form and function may occur with the associated micro flora in such environments. The present study was carried out to isolate and identify microbes in general and Bacillus spp. in particular from soil and sediment sample of Bhitarkanika mangrove ecosystem of Orissa coast to explore the extracellular enzymatic potential of the isolates. Generally Bacillus spp. are gram positive rods and resistant to high temperature. Soil and sediment samples were collected and was exposed to a temperature of 65°C for one hour in a water bath to remove undesirable soil bacteria and to isolate the heat resistant Bacillus spp. by dilution plate technique using Bacillus Hi-Chrome Agar. After Gram staining, spore staining and biochemical studies following Bergey’s Manual of Systematic Bacteriology (Pelczar,Chan & Krieg), it  was observed that the collected soil sample has a rich flora of Bacillus spp.. After biochemical test, the isolates were found to be Bacillus cereus, Bacillus subtilis, Bacillus sphaericus, Bacillus polymyxa, Bacillus macquariensis, Bacillus smithii, Bacillus licheniformis, and Bacillus pasteurii. All the isolates were screened for production of alpha-amylase and were found positive .Among all, B.smithii was the highest amylase producer i.e. 18.07 unit/ml at 55°C. After 4th day of incubation, the qualitative and quantitative assay of the enzyme was done following 3, 5-Dinitro salicylic acid method (Miller, 1959). Again the enzyme production was optimized at different pH, ‘C’ sources, and ‘N’ sources.

Keywords

Bacillus, Amylase, Biochemical identification, Qualitative and Quantitative assay

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