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Novel Approach of Alkaline Protease Mediated Biodegradation Analysis of Mustard Oil Driven Emulsified Bovine Serum Albumin Nanospheres for Controlled Release of Entrapped Pennisetum glaucum (Pearl Millet) Amylase

Kirti Rani, Shreya Goyal and Chanchal Chauhan

Amylase is known for starch saccharification and catalyses the starch hydrolysis producing glucose and limit dextrin which has wide range of applications in fermentation, textile, leather, detergents, food and paper industries. In the present work, amylase was extracted from Pennisetum glaucum (pearl millet) was entrapped in bovine serum albumin (BSA) nanoparticles in which emulsification was done by mustard oil and covalently coupled by using well known cross linking agents, glutaraldehyde. Bovine serum albumin is considered the most biodegradable, biocompatible and non-toxic biological matrix material for the preparation of enzyme bound nanospheres to enhance thermal stability, storage stability and suitability of bound enzymes as compared to free enzymes toward persisting ambient conditions. Formation of nanoparticles was confirmed by Dynamic Light scattering (DLS) and determination of exact size was observed by Scanning Electron Microscopy (SEM). Thermal stability of entrapped Pennisetum glaucum amylase at 70ºC and storage stability at 4ºC was enhanced for 3 hours 30 minutes and 12 months respectively as compared to free enzyme which was observed for 50 minutes and 1 day only. Studied optimal temperature of entrapped amylase was found to be 70ºC and it was significantly higher as compared to free enzyme (40ºC) which increased the industrial efficacy and catalytic viability of amylase after entrapment. Their biodegradation was carried out by using different units of alkaline protease (10U, 15U, 20U, 25U, 30U, 35U, 40U, 45U, 50U) to study sustained and controlled release of entrapped Pennisetum glaucum. Characterization of kinetic parameters were also performed for Pennisetum glaucum extracted amylase and entrapped amylase for their comparative study for their optimal pH, incubation time, substrate concentration, CaCl2 concentration and temperature for maximal enzyme activity which was tested by dinitrosalicylic acid method.