Production and Characterization of Lipase from Novel Bacterium Stenotrophomonas maltophilia RSP-09

Order of Publishing in Issue: 
10
Volume :9
Issue :1
February, 2015
Page No: 
75-87
Authors: 
Varanasi Venkata Naga Yaswanth, Jamal Ahmad, Buddana Sudheer Kumar, Reddy Shetty Shravani and Reddy Shetty Prakasham[*]
Address: 
Bio Engineering and Environmental Sciences, Indian Institute of Chemical Technology, Hyderabad, India
Email-ID: 
prakashamr@gmail.com

Lipase producing strains play key role in the enzymological remediation of oil polluted soils. Diversity of cultivable lipase producing bacteria in the industrial localities of Hyderabad was studied with samples collected from different areas. Among isolated bacterial strains, the highest lipase producing strain was identified both biochemically and 16S rRNA ribotyping. The 16S rRNA studies revealed that the strain belongs to Xanthomonadaceae family and identified as Stenotrophomonas maltophilia. An overall 3-fold enhanced lipase production (175μmoles/ml) was achieved after improved conditions of production medium. Among used nutrient sources sesame oil and tryptone were found to be the most suitable substrates for maximum enzyme production. The maximum enzyme production was observed at pH 8.0 and at rotation speed of 150 rpm. Na+ ions induced the enzyme production where as Hg2+ ions inhibited the enzyme production. 80 % Ammonium Sulphate precipitation showed maximum lipase activity and the molecular weight of produced lipase was identified 45 kDa based on SDS-PAGE. Optimum enzyme activity was noticed at pH (6.0) and at 50 oC. The observed Km for this enzyme is 0.727 and showed Vmax of 156.6μmoles/ml indicating lower Km than reported lipases that confirmed higher affinity of the enzyme for its substrate tributyrin. Arrhenius plot analysis for energy of activation denoted 11.07 Kjmol-1 K-1 for this lipase.

Keywords: 
Lipases, 16S rRNA, Stenotrophomonas maltophilia, Optimization, Ammonium sulphate precipitation, Enzyme kinetics, Activation energy.
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