IMMOBILIZED LIPASE - Toyobo

LIP-301TOYOBO ENZYMES(Industrial Grade)IMMOBILIZED LIPASEfrom Pseudomonas sp.PREPARATION and SPECIFICATIONThe enzyme is a preparation of a Pseudomonas sp. lipase (TOYOBO lipoprotein lipase GradeLPL-311) immobilized on Hyflo Super-Cel.Appearance: Light brown powder (immobilized on Hyflo Super-Cel)Activity: 0.5U/mg-solid or moreStabilizers: SugarsAPPLICATIONS 3,11This enzyme is useful for enzymatic ester synthesis, transesterification, acidolysis and alcholysis inorganic solvents or solvent free substrate.REFERENCES1) Biosci.Biotech.Biochem., 56 (7), 11181123 (1992)2) Agric.Biol.Chem., 53 (7), 18791884 (1989)3) J.Bull.Inst.Chem.Res., Kyoto Univ. 70 (3), 333337 (1992)4) Tetrahedron Letters, 31, No.25, 36033604 (1990)5) Chemistry Letters, 741744 (1993)6) Tetrahedron Letters, 35 No.43, 79978000 (1994)7) Tetrahedron:Asymmetry 6, No.7, 16171620 (1995)8) Tetrahedron 51 No.34, 93399352 (1995)9) Biosci. Biotech. Biochem., 59 (11), 21782180 (1995)10) Macromolecules 28, No.21, 70467050 (1995)11) Journal of Fermentation and Bioengineering 81 No.3, 206211 (1996)267

LIP-301ASSAYPrinciple:OOn-C 7 H 15 C-OCH 2 OF 3 C 2 H 5 OHimmobilized lipasen-C 7 H 15 C-OC 2 H 5 OF 3 CH 2 OHTFEO ethanol EOUnit definition:One unit causes the formation of the one micromole of ethyloctanoate (EO) per minute under the conditions describedbelow.Procedure1. Pipette 10.0ml of dry diisopropyl ether containing 50mM 2,2,2-trifluoroetyl octanoate (TFEO) and 0.2Methanol into a test tube with a magnetic stirrer bar and a stopper. Equilibrate at 25.5 for about 10minwith stirring.2. Add about 20mg of immobilized lipase powder and stir constantly.3. Take an aliquot of the assay mixture at a specific time interval and analyse with capillary gaschromatography (DB-5, J&W Scientific, 160). The retention time of TFEO and the product EO are 2.14 and2.54 min, respectively.4. A plot natural logarithm of the reaction conversion (C) vs reaction time gives a straight line In(1C)k 0 t, and the initial rate (V 0 ) of the reaction is calculated from the slope of the line.CalculationActivity can be calculated by using the following formulaInitial rate (V 0 ) k 0 50Weight activity (U/mg)V 0 10Wsk 0Slope of the lineIn(1C)k 0 t50 Concentration of TFEO (mM)Ws Sample weight (mg)10 Total volume (ml)268

LIP-301Illustrations of Stereoselective AcylationScheme1. Stereoselective Acylation of Binaphthol 2)OHOHOR-COimmobilized lipaseiPr 2 O 4.5mlacetone 0.5mlOOCROHOHOHCH 3CHO()402: binaphthol1: a R CH 3d nC 7 H 15b nC 3 H 73: binaphthol monoestersc nC 5 H 11Scheme2. Stereoselective Acylation of Mandelonitrile 3)OHOO-C CH 3 OHimmobilized lipaseCN CH 3 C-O CN CNO()1: 1-cyano-1-phenylmethanol (mandelonitrile)2: 1-cyano-1-phenylmethanol acetate3: isopropenyl acetateScheme3. Stereoselective Acylation of Tricarbonyl (o-trimethylsilylbenzyl alcohol) chromium 4)OH immobilized lipaseOHO AcSiMe 3OSiMe 3SiMe 3Cr(CO) 3 Cr(CO) 3Cr(CO) 3O(R.S)1: tricarbonyl (o-trimethylsilylbenzyl alcohol) chromium2: tricarbonyl (o-trimethylsilylbenzyl alcohol) chromium acetate269

LIP-301 7 15 2 3 2 5 7 15 2 5 3 2 2 P 0 0 0 0 immobilized lipase0 0 0 270