Purification and Characterization of a Novel Solvent-Tolerant Lipase from Fusarium heterosporum

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A microorganism producing a solvent-tolerant lipase was identified as Fusarium (F.) heterosporum. The lipase was purified from the culture filtrate to homogeneity as judged by disc-PAGE and SDS-PAGE. The purification included SP-Sephadex chromatography, gel filtration and isoelectric focusing, and the recovery yield was 38%. The lipase was a monomeric protein with a molecular weight of 31 kDa estimated by SDS-PAGE, and a pI of 7.0. The optimum pH at 40℃ and optimum temperature at pH 5.6 were 5.5-6.0 and 45-50℃, respectively, when olive oil was used as the substrate. The lipase was stable over a pH range of 4-10 at 30℃ for 4 h, and up to 40℃ at pH 5.6 for 30 min. Furthermore, the enzyme was not inactivated even after incubation at 30℃ in 50% solvent such as dimethylsulfoxide (DMSO), hexane, benzene and ether for 20 h. The activity did not decrease in a reaction with stirring in a mixture containing 50% DMSO or dimethylformamide. The lipase preferably reacted on middle-chain fatty acid triglycerides (6≤C≤12), and cleaved only 1,3-ester bonds of triolei. The enzyme had an N-terminal sequence of Ala-Val-Thr-Val-Thr-Thr-Gln-Asp-Leu-Ser, which has not previously been found in any other protein. We compared the properties of lipases from F. heterosporum and another strain F. oxysporum.
社団法人日本生物工学会の論文
1993-05-25