PURIFICATION AND CHARACTERIZATION OF A NEW ORGANIC SOLVENT TOLERANT LIPASE FROM CLADOSPORIUM CLADOSPORIOIDES SP. STRAIN NK-LF36N. KALYANIA , N. SARASWATHYB, S. BALAJIB AND P. RAMALINGAM BAbstract An extracellular lipase from Cladosporium cladosporioides was purified and characterized. The gelfiltration chromatography of crude extract yields purified lipase having the specific activity of 335.33 U/mg which was found to be 156.7-fold increase in yield with 51.75% recovery. Both the SDS-PAGE and Native- PAGE, showed a single band indicating enzyme molecular size of 62 kDa. The optimum pH and temperature of the enzyme was 6.0 and 30°C respectively. The enzyme retained 85% of the activity at 40°C and pH 7.5 respectively. Ca2+ and Mg2+ ions stimulated lipase activity. The enzyme retained 105% activity in n-butanol and more than 80% of the activity in toluene and n-hexane. The enzyme was specific for the substrate p-nitrophenyl palmitate, showing a low Km value of 0.82 mM and a Vmax value of 12.5 mM min-1. The lipase from NK-LF36 was thus characterized as mesophilic and solvent-tolerance proposes that it can be a potential catalyst in biodiesel production.
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