Abstract
A total of 20 bacterial isolates were obtained by screening using tributyrin agar medium. Among them the isolate which exhibited greater clearance zone and higher lipase activity was subsequently screened using spirit blue agar and rhodamine B agar medium. Based on morphological, biochemical and 16S rRNA sequence analysis, the potent isolate was identified as Staphylococcus aureus . The lipase production of the isolate was increased by improving the conditions of production medium. Maximum lipase production (8.11 U/ml) was achieved when 2% punnakka oil was utilized as sole carbon source at pH 7.0 and 37°C after 2 days of incubation. Addition of 3% tryptone as nitrogen source and 0.01% MgCl 2 resulted in a significant increase of lipase production (10.73 U/ml). The lipase production was slightly enhanced in the presence of 20% n-propanol and highly stable in the presence of n-butanol, toluene and n-hexane. The study resulted in isolation and production of inducible, mesophilic and solvent tolerant lipase with industrial potential. Keywords: Staphylococcus sp., tributyrin agar, 16S rRNA , medium optimization, solvent-tolerant lipase. African Journal of Biotechnology , Vol 13(28) 2858-2866
Highlights
Lipases catalyze the hydrolysis of ester linkages of triglycerides at water-oil interface (Gupta et al, 2011)
Microbiological media such as potato dextrose agar, tributyrin agar, spirit blue agar, rhodamine B and sodium deoxycholate were purchased from HiMedia Laboratories, Mumbai, India
Several methods have been proposed for screening of lipase production but there is still more scope for finding lipases with novel and specific properties through screening using tributyrin agar and plate assay (Rohit et al, 2001)
Summary
Lipases (triacylglycerol acylhydrolases, EC 3.1.1.3) catalyze the hydrolysis of ester linkages of triglycerides at water-oil interface (Gupta et al, 2011). Apart from hydrolysis, some lipases can catalyze reverse reactions including synthesis of esters by esterification, transesterification and interesterification (Franken et al, 2011). Existence of lipase producing microorganisms have been found in diverse habitats such as industrial wastes, vegetable oil processing units, dairies, soil contaminated with oil, compost heap, hot springs etc ( Sztajer et al, 1988; Wang et al, 1995). Bacterial lipases are mostly extracellular and are greatly influenced by nutritional and physico-chemical parameters such as temperature, pH, nitrogen and carbon sources
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