Isolation of Alkalitolerant and molecular characterization of their α-Amylase
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Date
2019
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Abstract
Alkalophiles are those organisms which can survive at high pH values roughly (9-11). The enzymes extracted from the alkalophiles are alkaline enzymes. Enzymes obtained from the microbial source are cheap, that’s why industries feel more convenience using enzymes that are from microbial source. These enzymes have now a days various applications in various industries. Microbial enzymes are used in paper industry, brewing industry, textile industry, pharmaceutical industry and food industry etc. In this study samples were collected from premium paper mills sheikupora road having pH 9. Isolated colonies were selected by inoculating the sample on LB broth and streaking the plate from the culture. This study includes identification of amylase producing bacteria, for this three tests were carried out on isolated bacteria, starch hydrolysis test, casein hydrolysis test and salt dilution test to check their ability to produce amylase enzyme. This study also included with the DNA extraction using organic extraction method, 16s rRNA amplification by using PCR and 16s rRNA sequencing. 16s rRNA sequence was matched 99.4% with the 16s region of the bacterium Vibrio cincinnateinsis ATCC35912. Phylogenetic analysis was performed for the characterization of the α amylase gene. The Phylogenetic tree for 16s rRNA sequence showed us that our isolated bacterium is closely related to Vibrio cincinnateinsis ATCC 35912 and distinctly related to the uncultured bacterium. The Phylogenetic tree for α-amylase gene depicted us that Vibrio cincinnateinsis NCTC 12012 is closely related to Vibrio neocalnidonicus strain and distinctly related to Citrobacter freundii. The phylogenetic tree for α-amylase protein also showed us that Vibrio cincinnateinsis NCTC 12012is closely related to Vibrio valnificus and distinctly related to Citobacter freundii.