title: Mitigation of acrylamide by l-asparaginase from Bacillus subtilis KDPS1 and analysis of degradation products by HPLC and HPTLC.
abstract: The use of bacterial l-asparaginase (LA) is one of the alternative approaches for acrylamide reduction in food stuffs as it catalyzes the conversion of l-asparagine to l-aspartic acid and ammonia. In present investigation, purification of extracellular LA from isolate of Bacillus subtilis sp. strain KDPS-1 was carried out by solid state fermentation process. The effects of solid substrates, initial moisture content, moistening agents, temperature, and incubation time on LA production was studied, and the highest asparaginase activity (47 IU/ml) was achieved in the medium having orange peel as substrate. The enzyme was purified to homogeneity by diethylaminoethyl (DEAE) cellulose ion exchange chromatography; with 84.89 % yield and 12.11 fold purity. LA showed stimulant activity against β-mercaptoethanol and was greatly inhibited by Zn(2+) and Hg(2+) metal ions. Reduction of acrylamide in fried potatoes was detected by high performance liquid chromatography, which showed clear degradation of acrylamide by height and area (%) in the chromatograms of standard sample to that of the test sample. Hydrolysates analysis by high performance thin layer chromatography confirmed the test sample to be LA.
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra University, Rajkot, 360005 India ; Max Planck Institute of Developmental Biology, Tubingen, Germany.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Sanghvi
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra University, Rajkot, 360005 India.
authors.forename: Kapil
authors.initials: K
authors.lastname: Bhimani
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra University, Rajkot, 360005 India.
authors.forename: Devendra
authors.initials: D
authors.lastname: Vaishnav
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra University, Rajkot, 360005 India.
authors.forename: Tejas
authors.initials: T
authors.lastname: Oza
authors.affiliation: Department of Biochemistry, Saurashtra University, Rajkot, India.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Dave
authors.affiliation: B. N. Patel Institute of Paramedical Sciences, Bhalej Road, Anand, India.
authors.forename: Prashant
authors.initials: P
authors.lastname: Kunjadia
authors.affiliation: Department of Pharmaceutical Sciences, Saurashtra University, Rajkot, 360005 India.
authors.forename: Navin
authors.initials: N
authors.lastname: Sheth
keywords: Acrylamide; HPLC; HPTLC; Solid state fermentation; l-Asparaginase