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Production, Purification and Application of Naringinase from soil isolates of Aspergillus Sp

Start Date: Tuesday, Jan 01,2013

End Date: Thursday, May 30,2013

School: School of Biotechnology

Thematic Area: Biotech

Project Incharge:Ajith Madhavan
Co-Project Incharge:Anjali M, Krishnendhu P Pray, Ramzyshihab ,S Pushpanjali, Acharya ,Sreerag V
Production, Purification and Application of Naringinase from soil isolates of Aspergillus Sp

Aspergillus sp. has the ability to produce Naringinase enzyme in a selective media under specific conditions. Naringinase has α-L-rhamnosidase and β-D-glucosidase activities. This hydrolytic enzymatic complex has wide occurrence in nature and has been reported in plants, yeasts, fungi and bacteria. Naringinase is commercially attractive due to its potential usefulness in pharmaceutical and food industries. It is of particular interest in the biotransformation of steroids, antibiotics and mainly glycoside hydrolysis. The glycoside, naringin containing terminal α-rhamnose and β-glucose which can act as substrates for naringinase.Naringin (4’,5,7’-trihydroxyflavanone-7-rhamnoglucoside) can be hydrolysed by α-L-rhamnosidase activity of naringinase to rhamnose and prunin (4’,5,7’-trihydroxyflavanone-7-glucoside) which can be further hydrolysed into glucose and naringenin (4’,5,7’-trihyroxyflavanone)by the β-D-glucosidase component of naringinase.
Here,Aspergillusnigergroup strains isolated from 25 different soil samples collected from various location of Kollam and Karunagapally, all the 25 samples were considered to be potential sources of Naringinase enzyme after the qualitative assay of the same in SSMM. All of the listed isolates were inoculated into the Naringin containing production media and incubated for 7 days under standard condition. The production media was checked for the enzyme activity using TLC and Davis method. TLC and HPLC method is qualitative in nature but Davis method is quantitative. Before performing HPLC, TLC was done with all the crude extract samples and the isolates which showed positive results were purified by Ammonium sulphate precipitation followed by dialysis. 5 isolates of 25 samples showed the enzyme yield which was used in the application of Naringinase. The study aimed to isolate the Aspergillus sp. which can yield higher amount of Naringinase enzyme. Using Davis method, the maximum activity of 0.122μg/min was obtained. The enzyme showed hydrolysis of Tannic acid to Gallic acid.

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