PRODUCTION AND PARTIAL-PURIFICATION OF A LIPASE FROM PSEUDOMONAS-PUTIDA 3SK

Abstract

Extracellular lipase was produced by the solvent-resistant strain Pseudomonas putida 3SK during the late logarithmic growth phase. The enzyme was purified 21-fold with 5.3% recovery by ion exchange chromatography and gel filtration chromatography. The molecular weight of the purified enzyme by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was shown to be 45 kDa. The lipase had an optimum pH range of 8.0-9.0 and was stable within the pH range 4.0-10.0. The optimal temperature was 37-degrees-C and it was stable up to 75-degrees-C. When the effects of organic solvents were tested, isooctane was most suitable for enzymatic fat hydrolysis in the two-phase system, Enzyme activity was inhibited by mercury ions and sodium dodecyl sulfate, whereas calcium ion and taurocholic acid stimulated its activity.