Abstract
Yarrowia lipolytica is capable of producing lipase, an enzyme widely used in industry for, under conditions microaquosas, catalyzing the synthesis of esters. The extracellular lipase produced by this yeast has been fully characterized and has wide range of application, but the intracellular and cell-bound fractions are still little reported production and characterization. The application of intracellular and cell-bound lipases in industrial processes can reduce the purification cost, facilitating the use of this enzyme. Therefore, the objective of this study was to evaluate the production of intracellular lipases induced by different carbon sources. We investigated the intracellular lipase production by Yarrowia lipolytica strain IMUFRJ 50682 using glucose, glycerol, crude glycerol, olive oil and frying oil. The methodology for the intracellular and cell-bound lipase extraction was standardized. Cell growth, concentration of glucose and glycerol was accompanied. Measures of enzymatic activity were carried out using spectrophotometric method. Y. lipolytica cells presenting in the form of hyphae were more sensitive to the two extraction processes used and the extraction with ultrasound possible to obtain activity values to cell bound fraction up to 9 times higher than those obtained with the mechanical extraction. Glycerol and glycerin were not effective for induction of enzymatic fractions. Glucose induced activity in extracellular fraction. Olive oil and waste frying oil were the best inducers for intracellular fractions. Frying oil was a good inductor to obtain intracellular lipases and the values of hydrolytic activity were similar to values obtained with the olive oil. In presence of frying oil, the hydrolytic activity was 12,19 U/g, in 24h of growth and when olive oil was used like inductor, the enzyme activity to the cell bound fraction reaching activity of 12,14 U/g in 24h of the experiment. Therefore, the intracellular fractions of Yarrowia lipolytica lipase can be induced efficiently and represent an interesting alternative for developing technology to the use of biomass in industrial processes.
