Purification and Characterization of a Novel Lipolytic Enzyme from Aspergillus oryzae

概要

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Aspergillus oryzae produced at least two kinds of extracellular lipolytic enzymes when cultured in a submerged culture medium containing 3% soybean oli. The enzymes, L1 and L2,exhibited higher substrate specificity towed dimercaptobutyrate and olive oil, respectively. The lipolytic enzyme (L1) was purified from the culture filtrate to homogeneity by ammonium sulfate and acetone precipitation, deithylaminoethyl (DEAE)-cellulose column chromatography, palmitoylated-gauze column chromatography, DEAE-Sephadex A-50 column chromatography and gel filtration, and was a monomeric protein with a molecular mass of 24 kDa estimated by gel filtraton and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme showed little prefernce for triacxylglycerols, and also exhibited high activities on ethyl esters of various fatty acids. The enzyme cleaved all the ester bonds of triolein. The optimum pH values when olive oil and tributyrin were used as substrates were 7.5 and 10.0,respecitively. One fragment derived from the enzyme cleaved off by cyanogen bromide had an N-terminal sequence of Asn-Gly-Ale-Ile-Lys-Arg-Leu-Ser-Ala-Asp-Val-Gln-Asp-Lys-Ile-Lys-Gly-Val-Val. Enzymatic properties and amino acid sequences of the lipoliytc enzyme (L1) from A. oryzae and cutinases from other fungi are compared and discussed.
社団法人日本生物工学会の論文
1994-12-25