Keratinase production by proteolytic micro-organisms in activated sludge and farmyard waste.
Okoroma, Emeka and Purchase, Diane and Garelick, Hemda and Abiola, Oduola (2008) Keratinase production by proteolytic micro-organisms in activated sludge and farmyard waste. In: Society for General Microbiology, Autumn Meeting, 8-11 September 2008, Trinity College, Dublin, Ireland.
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Thirty-two microbial strains from activated sludge and farmyard waste
were isolated on feather-meal agar. Five of these isolates demonstrated
significant proteolytic activity raising prospects of biotechnological
application. The keratinolytic activity was assessed by casein-agar plate
assay and further confirmed by keratinase assay using keratin azure as
substrate. One of the five isolates (a Gram-positive cocci bacterium)
produced the highest keratinase activity within 24h at a cell density of
5.2 x 108 CFU/mL under the optimized conditions of pH 10, 37°C and
substrate concentration of 0.4 g/L. The crude extract gave an activity
of 14 U/mL. Addition of carbon (starch) and nitrogen (yeast extract
and peptone) sources caused a decline in enzymatic activity. The
culture supernatants was purified by ultra filtration, concentrated by
Amicon Ultra-15 centrifugal filter device and further purified by
HiTrap affinity chromatography column. The resulting fraction has a
purification factor of 108.7 and specific activity of 2000 U/mg. The
molecular weight of the enzyme was determined by Matrix-assisted
laser desorption/ionization (MALDI) to be 27.9 KDa.
|Item Type:||Conference or Workshop Item (Poster)|
|Research Areas:||A. > School of Science and Technology > Natural Sciences|
|Depositing User:||Devika Mohan|
|Date Deposited:||02 Feb 2010 07:02|
|Last Modified:||06 Oct 2015 14:54|
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