NEHA LUHACH, NISHA SETHI, SANGITA YADAV, SUBHASH CHANDER, SWETA KUMARI AND ASHA GUPTA*
Department of Environmental Science and Engineering, Guru Jambheshwar University of Science and Technology,
Hisar-125 001 (Haryana), India
*(e-mail: guptaasha.phd@gmail.com; Mobile: 85128 65309)
(Received: January 10, 2023; Accepted: February 21, 2023)
ABSTRACT
Lipases enzyme leads to the fat hydrolysis into fatty acid and glycerol at the interface of lipid-water
along with the reactions leading to bioconversion in micro and non-aqueous surroundings. In this research, the conditions of culture required for the extracellular production of lipase by the native strain of bacteria isolated from the waste water and sludge samples collected from the sites near district Hisar, India were optimized. The optimization of lipase production was done using FCCCD of RSM. All optimized conditions temperature (55°C), pH (9), inoculum volume [3% (v/v)], agitation rate (200 rpm) and concentration of inducer oil [3% (v/v)] resulted in maximum lipase production of 256.20 µ/ml. The LipBL-WII(c) lipase produced by Bacillus licheniformis WII(c) strain was then subjected to sequential method in order to produce complete purification fold of 13.0%, recovery 30.4%, specific activity 760 µ/mg and a m.w. of 41 kDa. The pH and temperature optima for the LipBL-WII(c) were 8 and 55°C, respectively. Moreover, the lipase showed good steadiness amongst various surfactants, metal ions and organic solvents. The values for Km and Vmax were observed to be 1 mM and 6.66 mmoles, respectively. The purified lipase was used in the production of biodiesel. LipBL-WII(c) can be utilized for catalyzing the production of biodiesel using olive oil via transesterification reactions due to the higher lipolytic activity in thermophilic conditions and maximum stability in the existence of organic solvents.
Key words : Lipase, Bacillus sp., biodiesel, transesterification