Purification a d Characterization of from Aspergthus oryzae

tribute to the formation of the quality and function of miso. Lipases catalyze the hydrolysis of glycerides of soybeans and the synthesis of esters during miso fermentation,2) so that the fiavors and tastes are formed. However, there have been only a few reports about lipase from A. oryzae.3'`} Therefore, it is necessary to investigate the properties of these lipolytic enzymes and elucidate their roles to identify the mechanism of the change in lipids during mtso ferrnentation. A. or),zae produces at Ieast two extracellular lipolytic enzymes, Ll and L2, with diiierent substrate specificities.S) Both of these enzymes have been purified and characterized.6J) Ll, a cutinase-like enzyme, has a high specificity toward dimercaptobutyrate, and L2 hydrolyzes monoacylglycerols and diacylglycerols, but not triacylglycerols. Cloning and nucleotide sequencing of the genes coding for the enzymes has been done.8・9) We are interested in their substrate specificities, but triacylglycerol lipase, a third enzyme, seems to be the most important enzyme in its action during mtso fermentation. Triacylglycerol is the main lipid in soybeans. However, triacylglycerol ipase isolated from A. or "zae has not yet been reported. In a proj ect done to obtain enough of the purified protein for further study, we selected A. oryzae RIB128 as the most productive strain tested.iO) We also selected a suitable medium and culture conditiens for production of the lipase.'O) In this paper, we describe the purification and characterization of triacylglycerol lipase, designated L3.