Candida rugosa

Kumar and co-workers reported that N-arylaziridine-2-carboxylates could be resolved with the aid of lipase from Candida rugosa [78]. Moderate to high enantiose-... 

In this case study, an enzymatic hydrolysis reaction, the racemic ibuprofen ester, i.e. (R)-and (S)-ibuprofen esters in equimolar mixture, undergoes a kinetic resolution in a biphasic enzymatic membrane reactor (EMR). In kinetic resolution, the two enantiomers react at different rates lipase originated from Candida rugosa shows a greater stereopreference towards the (S)-enantiomer. The membrane module consisted of multiple bundles of polymeric hydrophilic hollow fibre. The membrane separated the two immiscible phases, i.e. organic in the shell side and aqueous in the lumen. Racemic substrate in the organic phase reacted with immobilised enzyme on the membrane where the hydrolysis reaction took place, and the product (S)-ibuprofen acid was extracted into the aqueous phase. 

Fig. 5.13. Lipase-catalysed hydrolysis of racemic ibuprofen ester. CRL Candida rugosa lipase.

Lipases from C. antarctica and P. cepacia showed higher enantioselectivity in the two ionic liquids l-ethyl-3-methylimidazolium tetrafluoroborate and l-butyl-3-methylimidazolium hexafluoroborate than in THE and toluene, in the kinetic resolution of several secondary alcohols [49]. Similarly, with lipases from Pseudomonas species and Alcaligenes species, increased enantioselectivity was observed in the resolution of 1 -phenylethanol in several ionic liquids as compared to methyl tert-butyl ether [50]. Another study has demonstrated that lipase from Candida rugosa is at least 100% more selective in l-butyl-3-methylimidazolium hexafluoroborate and l-octyl-3-nonylimidazolium hexafluorophosphate than in n-hexane, in the resolution of racemic 2-chloro-propanoic acid [51]. 

Very recently Page and coworkers have reported the D KR of sec-amines using a low catalyst loading of an Ir complex for the racemization, and Candida rugosa lipase for the enzymatic resolution [38]. 

Scheme 7.26 Kinetic resolution and DKR of 1-methyl tetrahydro-isoquinoline using Candida rugosa lipase.

The outstanding influence of the anionic component on the activity and selectivity of enzymes was demonstrated in the Candida rugosa lipase-catalyzed kinetic resolution of ibuprofen, a nonsteroidal antiinflammatory drug with sales of USD 183 million in 2006 (Scheme 5.15) [63]. 

We initially tested Candida antarctica lipase using imidazolium salt as solvent because CAL was found to be the best enzyme to resolve our model substrate 5-phenyl-l-penten-3-ol (la) the acylation rate was strongly dependent on the anionic part of the solvents. The best results were recorded when [bmim][BF4] was employed as the solvent, and the reaction rate was nearly equal to that of the reference reaction in diisopropyl ether. The second choice of solvent was [bmim][PFg]. On the contrary, a significant drop in the reaction rate was obtained when the reaction was carried out in TFA salt or OTf salt. From these results, we concluded that BF4 salt and PFg salt were suitable solvents for the present lipase-catalyzed reaction. Acylation of la was accomplished by these four enzymes Candida antarctica lipase, lipase QL from Alcaligenes, Lipase PS from Burkholderia cepacia and Candida rugosa lipase. In contrast, no reaction took place when PPL or PLE was used as catalyst in this solvent system. These results were established in March 2000 but we encountered a serious problem in that the results were significantly dependent on the lot of the ILs that we prepared ourselves. The problem was very serious because sometimes the reaction did not proceed at all. So we attempted to purify the ILs and established a very successful procedure (Fig. 3) the salt was first washed with a mixed solvent of hexane and ethyl acetate (2 1 or 4 1), treated with activated charcoal and passed into activated alumina neutral type I as an acetone solution. It was evaporated and dried under reduced... 

Finally, 2-keto-4-hydroxyalkanephosphonates 56 were resolved either by CAL-B-promoted acetylation or by lipase from Candida rugosa (CRL)-mediated hydrolysis of the corresponding 0-butyrates 57 (Scheme 4)7 ... 

Chloroform 2 OS-Phosphate buffer pH 7.2 (1/1) Candida rugosa lipase... 

Candida rugosa Physical adsorption on Ethyl laurate... 

Lipase 1 from Candida rugosa Esterification of 1-butanol and oleic acid Oil-deionized water (96.6/3.4) 91.4 103... 

Lipase 1 from Candida rugosa Transesterification 2-ethyl-l-hexanol and rapeseed oil Oil-water (97/3) 99.6 103... 

Membrane reactor Ethyl laurate hydrolysis Candida rugosa lipase 116... 

Membrane reactor Synthesis of monoglycerides Lipase from Candida rugosa 75... 

Hydroxy-isobutyric acid 2 Captopril Treatment for hypertension Hydroxylation Candida rugosa Single-stage fermentation [4]... 

Crawshaw, W.M., MacDonald, N.R. and Duncan, G. (2005), Outbreak of Candida rugosa mastitis in a dairy herd after intramammary antibiotic treatment . Veterinary Record, 156, 812-813. 

Dyal, A., Loos, K., Noto, M., Chang, S., Spagnoli, C., Shafi, K.V.P.M., Ulman, A., Cowman, M. and Gross, R.A. (2003) Activity of Candida rugosa lipase immobilized on y-Fe203 magnetic nanoparticles. Journal of the American Chemical Society, 125, 1684—1685. 

Layered phosphate/phosphonate and phosphonate materials, obtained by substitution of the phosphate moiety by phosphonate groups, display interesting tunable hydrophilic/organophilic properties for adsorption processes. When Candida rugosa lipase (CRL) is simply equilibrated with zirconium phosphate and phosphonate [135,136], immobilization was demonstrated to take place at the surface of the microcrystals. However, because lipase exhibits a strong hydrophobic character, its uptake by zirconium phosphate and phosphonate was much more related to the hydrophobic/hydrophilic character of the supports than to the surface area properties. A higher uptake is observed for zirconium-phenylphosphonate (78 %)... 

More recently, ionic liquids have also been used as additives. Lee and coworkers explain that ionic liquids may ad as a template during the sol-gel process redudng the shrinkage of the matrix by pore filling and behave as a stabilizer to proted Candida rugosa lipases [184]. 

R. J. Kazlauskas, A. N. E. Weissfloch, A. T. Rappaport, L. A. Cuccia, A Rule to Predict Which Enantiomer of a Secondary Alcohol Reacts Faster in Reactions Catalyzed by Cholesterol Esterase, Lipase from Pseudomonas cepacia, and Lipase From Candida rugosa, J. Org. Chem. 1991, 56, 2655 - 2665. 

CCL - now known as lipase from Candida rugosa (CRL)) but not porcine pancreatic lipase (PPL) (Figure 1.12). 

Dalmau, E., Montesinos, J.L., Lotti, M. and Casas, C., Effect of different carbon sources on lipase production by Candida rugosa. Enzyme Microb. Technol., 2000, 26, 657-663. 

Brocca, S., Schmidt-Dannert, C., Lotti, M., Alberghina, L. and Schmid, R.D., Design, total synthesis, and functional overexpression of the Candida rugosa lipl gene coding for a major industrial lipase. Protein Sci., 1998, 7, 1415-1422. 

Enzymatic Resolution of 1-Methyl-tetrahydroisoquinoline using Candida rugosa Lipase... 

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