ALKALINE PROTEASE PRODUCTION USING SUBMERGED FERMENTATION FROM DIFFERENT SPECIES OF BACILLUS
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Date
2016
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Department of Molecular and Cellular Engineering Jacob School of Biotechnology and Bioengineering Sam Higginbottom Institute of Agriculture, Technology and Sciences, Allahabad-211007
Abstract
Alkaline protease is the most important group of protease enzymes utilized commercially in
various arenas of industries, such as food, detergent, leather, dairy, pharmaceutical, diagnostics,
and waste management. Microorganisms of specially those belonging to Bacillus genera serve
as a vast repository of diverse set industrially important enzymes and utilized for the large scale
enzyme production using fermentation technology.
Bacteria were isolated from soil samples from Lucknow in India using serial dilution method.
Different isolates were then screened for their alkaline protease producing capability using skim
milk hydrolysis test..Isolates showing positives result for the skim milk hydrolysis test were
identified based on their morphology by staining and their biochemical properties. DNA was
extracted from all the isolate using Phenol:chloroform method. Quantification of the isolated
DNA was performed by UV-Vis double beam spectrophotometer. The high yield DNA samples
were amplified using 16S rRNA primers and based on bioinformatic analysis thus identified five
different Bacillus species (Bacillus megaterium, Bacillus pumilus, Bacillus subtilis ,Bacillus
chonii and Bacillus cereus) were identified.
Eight fermentation media were used for production of Alkaline protease for identifying the
media in which yield is maximum for the Alkaline protease enzyme production.Media (MD8)
produced highest amounts of alkaline protease (62.5 U/mL).
The study also aimed at determining the most optimized conditions for Alkaline protease
production and activity from five different species of Bacillus like pH, metal ions, nitrogen
source, carbon source and NaCl. The most optimized media and culture conditions will help to
enhance the protease production which could play major role in various detergent industries.
Description
Ph.d thesis
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