Purification and Characterization of a New Lipase fromFusariumsp. YM-30

Abstract

The extracellular lipase from Fusarium sp. YM-30 was purified by a procedure involving ultrafiltration, ammonium sulfate precipitation, and DEAE-Toyopearl 650M, CM-Toyopearl 650M, and Butyl-Toyopearl 650M column chromatographies. The purified lipase was homogeneous with 12kDa of molecular mass by SDS-PAGE, and had high specificities for mono- and diacylglycerols, but low toward triacylglycerols. The enzyme had maximum activity at pH 7.0 to 8.0 and 37 degrees C, and hydrolyzed digalactosyl diglyceride too.

Production, purification, characterization, and applications of lipases

“…roqueforti IAM7268. The lipase activity was not affected by Ca 2 + , Mg 2 + , Mn 2 + , Na + , K + , Cu 2 + , EDTA, p-chloro mercuribenzoic acid, and iodoacetate (Mase et al, 1995 (Lin et al, 1996). Metal chelators (EDTA, o-phenanthrolin) did not significantly affect the alkaline lipase activity (Lin et al, 1996).…”

“…roqueforti IAM 7268 was purified to homogeneity by a procedure involving ethanol precipitation, ammonium sulfate precipitation, and three chromatographic steps on different matrices (DEAE-Toyopearl 650 M, Phenyl Toyopearl 650 M, Toyopearl HW-60). The molecular mass of purified lipase was 25 kDa by electrophoresis (Mase et al, 1995). The enzyme had a high specificity towards short-chain fatty acid esters (Mase et al, 1995).…”

“…The molecular mass of purified lipase was 25 kDa by electrophoresis (Mase et al, 1995). The enzyme had a high specificity towards short-chain fatty acid esters (Mase et al, 1995). A Pichia burtonii lipase was purified to homogeneity by a combination of DEAE-Sephadex A-50 ion exchange chromatography, Sephadex G-100 gel filtration, and isoelectric focusing (Sugihara et al, 1995).…”

“…Effect of metal ions and chelating agents on lipase activity Chartrain et al (1993) observed that an extracellular lipase of P. aeruginosa MB5001 was strongly inhibited by 1 mM ZnSO 4 (94% inhibition) but was stimulated by adding 10 mM CaCl 2 (1.24-fold stimulation) and 200 mM taurocholic acid (1.6-fold stimulation). Mase et al (1995) studied the effect of metal ions (1 mM concentration) on a purified lipase of Pe. roqueforti IAM7268.…”

Cloning, Characterization, and Expression of cDNA Encoding a Lipase fromKurtzmanomycessp. I-11

“…9,20) The standard reaction mixture was composed of 0.95 ml of substrate (2.63 mM p-nitrophenyl laurate (Sigma, USA) in 50 mM acetate-Na buŠer, pH 5.6, containing 4z Triton X-100) and 50 ml of lipase Since p-nitrophenyl laurate is unstable at basic pHs, the pH-activity of lipase was measured bỳ`T riglyceride G Test Wako'' (Wako Pure Chemical Industries, Japan) using 1-monooleyl glycerol (Sigma, Approx.99z, USA) as substrate in various buŠers. 21,22) The standard assay mixture contained 5.6 nmol of substrate, 200 ml of n-hexane, 800 ml of 100 mM buŠer, and 100 ml of lipase solution. After reaction at 379 C for 30 min with mixing every 3 min, 1 ml of CHCl3 was added to stop the reaction and the glycerol released into the water layer was measured with a Triglyceride G Test Wako (Wako Pure Chemical Industries, Japan).…”