Candida cylindracea lipase production

The group of Williams has reported a DKR of halo a-bromo [33a] and a-chloro esters [33b]. In the latter case, the KR is catalyzed by commercially available cross-linked enzyme crystals derived from Candida cylindracea lipase. The racemization takes place through halide SN2 displacement. The DKR is possible because the racemization of the substrates is faster than that of the products (carboxylates). For the ester, the empty ( =0) orbital is able to stabilize the SN2 transition state by accepting electron density. However, the carboxylate is more electron-rich and therefore less able to facilitate an SN2 reaction. Racemization is performed by the use of a chloride source. The best results where obtained by using a resin-bound phosphonium choride (Scheme 5.17). 

Biocatalysts such as Candida cylindracea lipase enzymes have recently been used in enantioseleetive hydrolyses of racemic esters by Fowler, Maefarlane and Roberts.15 The (R,R) diastereoisomer was the main product (91 +/- 0.5 %) as determined by NMR spectroscopic analysis of the products. These authors point out the difficulty of measuring such diastereoisomer ratios by using NMR methods, however, their results show a clear selectivity towards the (R,R) diastereoisomer. 

Lara, P. V., and Park, E. Y. 2004. Potential application of waste activated bleaching earth on the production of fatty acid alkyl esters using Candida cylindracea lipase in organic solvent system. Enzyme and Microb. Technol., 34, 270-277. 

Nakano, H., Kitahata, S., Shimada, Y, Nakamura, M., Tominaga, Y, and Takenishi, S. 1995. Esterification of glycosides by a mono- and diacylglycrol lipase from Penicillium camembertii and comparison of the products with Candida cylindracea lipase. J. Ferment. Bioeng. 80 24—29. 

Moreover, the enantioselective synthesis of ibuprofen esters catalyzed by Candida cylindracea lipase was also studied in a similar system [115]. The enanti-oselectivity displayed by the lipase was much higher in AOT/isooctane microemulsions than in isooctane. The use of lipases to prepare large quantities of an optically enriched product has also been demonstrated in CTAB-based microemulsions [89]. Resolution of ( )-menthol by C. cylindracea lipase-catalyzed esterification with propionic anhydride in a nonionic reverse micellar system in an ultraiiltration manbrane reactor has been recently described [117]. 

Dynamic resolution reactions couple the in situ racemization of a starting material with a kinetic resolution, thereby providing a route for the conversion of both enantiomers of starting material into one enantiomer of product. However, there is an important caveat the reaction product must be essentially inert to racemization. In control studies, 2-phenylpropionic acid (224 R = H) has been shown to racemize only slowly when treated with DABCO (l,4-diazobicyclo[2.2.2]octane) in water-DMSO (1 19) at 40 °C, whereas its phenyl ester (224 R = Ph) treated similarly was almost completely racemized in 7h. This work led to a procedure employing Candida cylindracea lipase (CCL) in which D,L-2-phenylpropanoic acid underwent a promisingly efficient dynamic kinetic resolution (Scheme 40). ... 

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. 

In an enzymatic resolution approach, chiral (+)-tra .s-diol (60) was prepared by the stereoselective acetylation of racemic diol with lipases from Candida cylindraceae and P. cepacia. Both enzymes catalyzed the acetylation of the undesired enantiomer of racemic diol to yield monoacetylated product and unreacted desired (+)-trans-diol (60). A reaction yield of 40% and an e.e. of >90% were obtained using each lipase [104],... 

Kim, B. S., and Hou, C. T. 2006. Production of lipase by high cell density fed-batch culture of Candida cylindracea. Bioproc. Biosys. Eng., 29, 59-64. 

Lipase (Candida rugosa formerly Candida cylindracea) Produced as an off-white to tan powder by controlled fermentation using Candida rugosa. Soluble in water but practically insoluble in alcohol, in chloroform, and in ether. Major active principle lipase. Typical applications hydrolysis of lipids, manufacture of dairy products and confectionery goods, development of flavor in processed foods. 

Optically pure inositol intermediates are very useful for synthesising D-myo-inositol 1,4,5-trisphosphate, which was found to be a second messenger. Ozaki et al [73] studied the resolution of racemic di-O-cylohexylidene-myo-inositol derivatives 17 and 18 by lipase-catalyzed esterification in organic solvents. Lipase from Candida cylindracea exclusively acetylates the hydroxyl group at C-4 or at C-5 of the D-enantiomer of 17 or 18, respectively. Around 100% e.e s of monoacetate products and unreacted starting compounds were obtained. The efficiency of the resolution is affected by the solvent, the most hydrophobic solvents ethyl ether and benzene being more effective than the water miscible solvents such as acetone, THF, and dioxane. 

The possible application of enzyme-assisted reactions for production of lower value nonspecialty lipids such as margarine hardstocks and cooking oils has been reported (50). When nonspecific lipases, such as those from Candida cylindraceae and C. antarctica, are used as biocatalysts for interesterification of oil blends, the TAG... 

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