Crystallization and characterization of monoacylglycerol and diacylglycerol lipase from Penicillium camembertii

Abstract

A new lipase from Penicillium camembertii U-150, which is specific for monoacylglycerols and diacylglycerols, but not triacylglycerols, was purified as four active components using concanavalin-A -Sepharose column chromatography, crystallized in the form of needles, and its properties investigated. No significant difference was observed in substrate specificity, but molecular mass and other enzymatic properties, such as pH, heat stability and optimum pH and temperature, were clearly different between the unadsorbed and the three adsorbed components on concanavalinA -Sepharose ; the three adsorbed components were similar to each other and more stable than the unadsorbed component. On the other hand, after enzymatic removal of carbohydrates from the three adsorbed components, their enzymatic properties became similar to those of the unadsorbed component. The carbohydrates of this lipase contribute to the stability of the enzyme, but not to its enzyme activity. The amino acid compositions of the four components did not differ from each other, and tryptic mapping of the deglycosylated components and amino acid composition of the tryptic fragments were identical. The carbohydrate compositions of four intact components were, however, different from each other. All four components have the same polypeptide backbone and multiple forms of this lipase are due to the differences in composition of the carbohydrates bound in this lipase.

Efficient Expression of Mono- and Diacylglycerol Lipase Gene fromPenicillium camembertiiU-150 inAspergillus oryzaeunder the Control of Its Own Promoter

“…As reported previously, 18) the glycosylated form of native MDGL, which is a main component, has an apparent molecular mass of 39 kDa (Fig. 3, lane 1) and the unglycosylated form has that of 37 kDa on SDS-PAGE (lane S).…”

Potencial de biocatálise enantiosseletiva de lipases microbianas

“…Em relação as cepas de Penicillium, vários trabalhos foram encontrados envolvendo produção, caracterização e purificação de lipases de diferentes espécies, como P. citrinum 36 , P. cyclopium 37 , P. simplicissimum 38 , P. caseicolum 39 , P restrictum 40 , P. expansum 41 , P. chrysogenum 42 , P. roqueforti 43 , P. camembertii 44 , P. abeanum 45 , entre outras espécies 46 . Entretanto, parece que este é o primeiro relato sobre a produção de lipase pelo fungo Penicillium solitum.…”

Purification and Characterization of a Lipase fromAspergillus oryzae

“…The third type, mono-and diacylglycerol lipase from Penicillium camembertii U-ISO, was reported by Yamaguchi et al 17) and Isobe et at. 20) It hydrolyzed monoacylglycerols and diacylglycerols, but not triacylglycerols (C 2 -C 18)' Judging from the substrate specificity, the lipase from A. oryzae was different from the enzyme from rat adipose tissue in acting on both mono-and diacylglycerols, also from the enzyme from P. cyclopium in not acting on triacylglycerols, and had a similar enzyme activity toward mono-, di-, triacylglycerols to the lipase from P. camembertii.…”