Lipase Activity Microbial

Lawrence, R. C. 1967. Microbial lipases and related esterases. Part II. Estimation of lipase activity. Characterization of lipases. Recent work concerning their effect on dairy products. Dairy Sci. Abstr. 29, 59-70. 

Thomson, C. A., Delaquis, P.J., and Mazza, G. 1999. Detection and measurement of microbial lipase activity A review. Crit. Rev. Food Sci. Nutr. 39 165-187. 

The use of extracellular lipases of microbial origin to catalyze the stereoselective hydrolysis of esters of 3-acylthio-2-methylpropionic acid in an aqueous system has been demonstrated to produce optically active 3-acylthio-2-methyl-propionic acid [41-43], The synthesis of the chiral side chain of captopril by the lipase-catalyzed enantioselective hydrolysis of the thioester bond of racemic 3-acetylthio-2-methylpropionic acid (15) to yield 5 -(-)-(15) has been demonstrated [44], Among various lipases evaluated, lipase from Rhizopus oryzae ATCC 24563 (heat-dried cells), BMS lipase (extracellular lipase derived from the fermentation of Pseudomonas sp. SC 13856), and lipase PS-30 from Pseudomonas cepacia in an organic solvent system (l,l,2-trichloro-l,2,2-tri-fluoroethane or toluene) catalyzed the hydrolysis of thioester bond of undesired enantiomer of racemic (15) to yield desired S-(-) (15), R-(+)-3-mercapto-2-methylpropionic acid (16) and acetic acid (17) (Fig. 8A). The reaction yield of... 

Veeraragavan, K. 1990. A simple and sensitive method for the estimation of microbial lipase activity. Anal. Biochem. 186, 301-305. 

Application and Principle This procedure is used to determine the lipase activity in preparations derived from microbial sources and animal pancreatic tissues. The assay is based on the potentiometric measurement of the rate at which the preparations will catalyze the hydrolysis of tributyrin. 

Form Supplied in usually a white or brownish powder, but also immobilized on an appropriate support. Lipases from microbial sources are virtually homogeneous in terms of hydrolytic activity, while mammalian and plant lipase preparations contain several interfering enzymes including proteases and esterases. 

The microbial lipolytic activity is determined by comparing the rate at which a solution of microbial lipase powder hydrolyzes a substrate of olive oil emulsion with the rate at which a solution of microbial lipase F.I.P. standard hydrolyzes the same substrate under the same conditions. It should be clearly stated that a lipase of microbial origin is involved to prevent confusion with the assay for pancreatic lipase. 

Schoevaait R, Wolbers MW, Golubovic M et al. (2004) Preparation, optimization and structures of cross-Unked enzyme aggregates (CLEAs). Biotechnol Bioeng 87 754-762 Schulz T, Plesis J, Schmid RD (2000) Stereoselectivity of Pseudomonas cepacia lipase toward secondtuy alcohols a quantitative model. Protein Sci 9 1053-1062 Secundo F, Cturea, G, Tarabiono C et al. (2006) The lid is a stmctural and functional determinant of lipase activity and selectivity. J Mol Catal B Enzym 39 166-170 Seitz EW (1974) Industrial application of microbial lipases a review. JAOCS 51(2) 12-16 Sharma R, Chisti Y, Baneijee UC (2(X)1) Production, purification, characterization and application of lipases. Botechnol Adv 19 627-662... 

Johnstone B (1990) A throw away answer. Far Eastern Econ Rev 147 62-63 Joseph B, Ramteke PW, Thomas G (2008) Cold active microbial lipases some hot issues and recent developments. Biotechnol Adv 26 457-470 Jung K, Hany R, Rentsch D, Stomi T, Egh T, Witholt B (2000) Characterization of new bacterial copolyesters containing 3-hydroxyoxoalkanoates and acetoxy-3-hydroxyalkanoates. Macromolecules 33 8571-8575... 

Carbonyls are quite important to the flavor of cheeses. The TNO-CIVO compilation of volatiles in foods lists 29 carbonyls as having been identified in Cheddar cheese at that time. Carbonyls (methyl ketones) may arise in fermented prodncts iiutiaUy via lipase activity of the starter cnlture. Dairy products contain a significant quantity of a-keto acids which are readily hydrolyzed from the triglyceride by microbial lipases and then decarboxylated to form odd carbon nnmber methyl ketones. 

The mechanism of lipase activity is described under section 3.7.1.1. Lipase from microbial sources (e. g. Candida lipolytica) is utilized for enhancement of aromas in cheesemaking. Limited hydrolysis of milk fat is also of interest in the production of chocolate milk. It enhances the milk character of the flavor. 

Wina E, Muetzel S, Hoffinann E, Makkar HOS, Becker K (2005) Saponins containing methanol extract of Sapindus rarak affect microbial fermentation, microbial activity and microbial community structure in vitro. Anim Feed Sd Technol 121 159-174 Wright DE (1961) Bloat in cattle. XX. Lipases activity of rumen microorganisms. NZ J Agric Res 4 216-223... 

Persson, M., Costes, D., Wehtje, E. and Adlercreutz, P. (2002) Effects of solvent, water activity and temperature on lipase and hydroxynitrile lyase enantioselectivity. Enzyme and Microbial Technology, 30, 916-923. 

The stability of the ester surfactants against enzymatic hydrolysis by two different microbial Upases, Mucor miehei lipase (MML) and Candida antarc-tica lipase B (CALB) added separately to the surfactant solutions, was also investigated, see Fig. 5 [19]. It is obvious that hydrolysis of the unsubstituted surfactant is much faster with both CALB and MML than that of the substituted surfactants, i.e., increased steric hindrance near the ester bond leads to decreased hydrolysis rate. Since the specificity of the enzyme against its substrate is determined by the structure of the active site, it can be concluded, as expected, that the straight chain surfactant most easily fits into the active site of both enzymes. 

Lipases, which are noted for their tolerance of organic solvents, were obvious candidates for biocatalysis in ionic liquids. Indeed, stable microbial lipases, such as CaLB [8, 54, 55, 56] and Pseudomonas cepacia lipase (PcL) [28, 55, 57] were cat-alytically active in the ionic liquids of the l-alkyl-3-methylimidazolium and 1-alkylpyridinium families, in combination with anions such as [BF4], [PF6], [TfO] and [ Tf2N]. Early results were not always consistent, which may be caused by impurities that result from the preparation of the ionic liquid. Lipase-mediated transesterification reactions (Figure 10.3) in these ionic liquids proceeded with an efficiency comparable to that in tert-butyl alcohol [8], dioxane [57], or toluene... 

The cutinase from Fusarium solani pisii maintained its transesterification activity in [BMIm][BF4], [OMIm][PF6] and [BMIm][PF6] (in order of increasing activity) at aw=0.2 [59]. Candida rugosa lipase (CrL), which is generally much less tolerant of anhydrous media than other microbial lipases, has successfully been used in anhydrous as well as water-saturated ionic liquids [60, 61, 62, 63, 64, 65]. 

If a liquid form of the enzyme is added, such as lipases in broth samples from microbial cultures, calculate specific activity on a normalized basis by substituting a ml enzyme sample added (instead of mg protein) using the equation above. 

Enantioselective enzymatic transesterifications have been used as a complementary method to enantioselective enzymatic ester hydrolyses. The first example of this particular type of biotransformation is the synthesis of the optically active 2-acetoxy-l-silacyclohexane (5 )-78 (Scheme 19). This compound was obtained by an enantioselective transesterification of the racemic l-silacyclohexan-2-ol rac-43 with triacetin (acetate source) in isooctane, catalyzed by a crude lipase preparation from Candida cylindracea (CCL, E.C. 3.1.1.3)62. After terminating the reaction at 52% conversion (relative to total amount of substrate rac-43), the product (S)-78 was separated from the nonreacted substrate by column chromatography on silica gel and isolated in 92% yield (relative to total amount of converted rac-43) with an enantiomeric purity of 95% ee. The remaining l-silacyclohexan-2-ol (/ )-43 was obtained in 76% yield (relative to total amount of nonconverted rac-43) with an enantiomeric purity of 96% ee. Repeated recrystallization of (R)-43 led to an improvement of enantiomeric purity by up to >98% ee. Compound (R)-43 has already earlier been prepared by an enantioselective microbial reduction of the l-silacyclohexan-2-one 42 (see Scheme 8)53. The l-silacyclohexan-2-ol (R)-43 is the antipode of compound (.S j-43 which was obtained by a kinetic enzymatic resolution of the racemic 2-acetoxy-l-silacyclohexane rac-78 (see Scheme 15)62. For further enantioselective enzymatic transesterifications of racemic organosilicon substrates, with a carbon atom as the center of chirality, see References 64 and 70-72. 

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