ISOLATION, PARTIAL PURIFICATION AND CHARACTERIZATION OF A NOVEL THERMOSTABLE LIPASE FROM SERRATIA MARCESCENS SCL1
Journal Title: European Journal of Biomedical and Pharmaceutical Sciences - Year 2019, Vol 6, Issue 8
Abstract
Thermostable lipases have become challenge for wide application in modern industrialization. The present study aims in the isolation, partial purification and characterization of a thermostable lipase producing bacterial strain, Serratia marcescens scl1 from medicinal waste. The bacteria isolated on Tributyrin agar plates was characterized by different biochemical tests and confirmed by 16s-rDNA sequencing. The growth parameters of bacteria showed best growth at 350C and pH 7.0 with 1% olive oil as carbon source after 48 hours of incubation. The bacterial growth curve was analyzed and lipase partially purified taking 24 hour old cultures. The lipase production analyzed with Tween 20 and Tween 80 agar plates and quantitative assay of lipase activity carried out using 2.0 mM pNPP as substrate in 50.0mM Tris-HCl, measuring absorbance at 410nm. The optimum temperature and pH of extracellular lipase produced by the bacteria occurred at 750C and pH 8.0. The substrate saturation kinetics showed maximum at 1.3mM [S] and activity 5.43±0.05 X10-2 unit/ml. The protein concentration determined is 240 μg/ml and specific activity of the enzyme is 22.58 unit/mg. The results indicate the isolate are able to produce low cost high profile enzyme which can satisfy the requirements of future goal to modern industrialization.
Authors and Affiliations
Dr. Sibani Sen Chakraborty
Keywords
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