PURIFICATION AND CHARACTERIZATION OF LIPASE FROM MICROBIAL ISOLATES AND ITS APPLICATION IN FORMULATION OF FUNCTIONAL FOOD PRODUCTS
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Date
2019-04
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UHF,NAUNI
Abstract
ABSTRACT
Enzymes are considered as nature’s catalysts. are a class of enzymes which catalyze the
hydrolysis of long chain triglycerides into polar lipids. Soil contaminated with oil from different sites of
Himachal Pradeshbeing a highly probablesource for lipolytic microorganisms was utilized for isolation of
lipase producing microorganisms. In total, 53 lipolytic bacteria have been screened on tributyrin agar
medium. Among them hyper lipolytic strains C6 and G7 were selected and identified asBacillus siamensis
C6MF446911 and Paenibacillus sp.G7. Cultural conditions and process parameters viz. media types,
pH, temperature, inoculum size, incubation time, substrate concentration, divalent ions and surfactants etc.
were optimized firstly through classical onevariable at a time (OVAT)followed by statistical optimization
by employing central composite design of response surface methodology. The enzymes obtained from
both the strains were purified to homogeneity by following a sequential purification approach. B.
siamensis C6 lipase was purified to a final purification fold of 1.91 and had a molecular weight of 32kDA
whereas P. sp.G7 lipase was purified to a final purification fold of 2.01 and had a molecular weight of 66
kDa. Lipase activity was found to be maximum at 40oC and pH 8.0 for B. siamensis C6 and at 35 oC and
pH 8.0 for P.sp.G7. Lipase from both the strains was quite thermostable with retention of more than 50%
activity after incubation of 120 min at 30-45 oC. Shelf life of lipase from both the strains showed that the
enzyme was very efficient qualitatively as well as quantitatively. Press cake of Brassica nigra showed
maximum stability for both isolates i.e. 1213.06 IU/mg specific activity for B. siamensis C6 lipase and
1236.84 IU/mg specific activity for P. sp.G7 lipase. Chitosan beads and κ-carrageenan chips at a
concentration of 1.5% and 4 % respectively, were found to be most efficient for immobilization of purified
lipase of P. sp.G7. Purified P. sp.G7 lipase showed a considerably good stability in κ-carrageenan chips
followed by immobilized chitosan beads (1.5%) giving an outstanding immobilization efficiency of 80.99
and 76.96% at the end of 20thcycle. Applicability of purified lipase from P. sp.G7 (20 ppm) in order to
enhance product quality and earn a natural and clean label status to bread which is highly sought after by
health conscious consumers of modern age. In addition pseudocereal grains. Therefore incorporation of
these grains in bread dough formulation enhances significantly the nutritional quality of baked product.
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