Cold-active lipase from Psychrobacter alimentarius ILMKVIT and its application in selective enrichment of ω-3 polyunsaturated fatty acids in flax seed oil.

Abstract

Lipases are one of the ubiquitous enzymes that belong to the hydrolases family and have a wide variety of applications. Cold-active lipases are of major attraction as they can act in lower temperatures and low water conditions because of their inherent greater flexibility. One of the novel applications of lipase is the enrichment of ω-3 polyunsaturated fatty acids (PUFA) in plant and fish oils. This study is aimed at the isolation and identification of cold-active lipase producing bacterium from marine sources, preliminary optimization of medium constituents and conditions, purification of lipase using chromatographic techniques, biochemical characterization, and ultimately the exploration of its application in the enrichment of ω-3 PUFA in flax seed oil. Psychrobacter alimentarius ILMKVIT was identified as the potential cold-active lipase producing bacterium based on its lipolytic activity in rhodamine B agar, titrimetric, and p-nitrophenyl palmitate (p-NPP) assays. One factor at a time (OFAT) analysis, revealed, an incubation time of 4.5 days, alkaline pH of 9, the temperature of 25 °C, peptone, and yeast extract as nitrogen sources, olive oil as inducer sources, 1% inoculum size, and NaCl as mineral sources as optimum production medium constituents and conditions for lipase production. Lipase purification was achieved by ion exchange and gel-filtration chromatography with a 9.27% yield and 37.51-fold purification. Biochemical characterization reported that the lipase is cold-active, alkaline, enhanced by Fe³⁺ metal ions, and tolerant to organic solvents, detergents, and inhibitors. P. alimentarius ILMKVIT lipase-hydrolysis followed by urea complexation of flax seed oil resulted in the enrichment of ω-3 PUFA, especially α-linolenic acid (ALA). Hence, the novel cold-active lipase from P. alimentarius ILMKVIT could be used to enrich ω-3 PUFA in flax seed oil and developed further as a prominent nutrient supplement for health benefits.