Presence Of Candida Rugosa Lipase Research Articles

Enzymatic synthesis of hydrophilic phytosterol polyol esters and assessment of their bioaccessibility and uptake using an in vitro digestion/Caco-2 cell model

A novel enzyme-catalyzed method was developed for the synthesis of phytosterol polyol esters from β-sitosterol and polyols (sorbitol, mannitol and xylitol) by two-step transesterification using divinyl adipate (DVA) as a link. A high conversion (exceeding 94%) of β-sitosterol with a vinyl group was achieved, in the presence of Candida rugosa lipase (CRL), at low temperature (35 °C) within 30 min. Subsequently, the maximum conversion of phytosterol polyol esters (>94%) was obtained using alkaline protease from Bacillus subtilis at 65 °C. Phytosterol polyol esters had enhanced thermal stability (up to an above 355 °C) and excellent water solubility (4.6–7.9 mM at 35 °C). Moreover, obvious increases in the bioaccessibility (41.5–63.6%) and intestinal uptake (5.2–6.5%) were observed using a simulated gastrointestinal digestion/Caco-2 cell model. These results highlighted the key role of hydrophilic structural modifications on physicochemical properties and absorption of phytosterols.

G_Fe3O4/Ag supporting Candida rugosa lipase for the “green” synthesis of pomegranate seed oil derived liquid wax esters

Wax esters were obtained through hydrolysis/esterification of pomegranate seed oil (PSO) with cetyl alcohol in the presence of Candida rugosa lipase (CRL) immobilized on G_Fe3O4/Ag@OA (oleic acid-coated magnetic nano-hybrid) catalyst. Lipase was immobilized successfully on the as-produced nanoparticles with an efficiency, depending on the operating conditions, up to ∼94%. The immobilized CRL shows very high activity on the respect of the free counterpart, which can be attributed to the lipase conformational structure modification related to interfacial activation. The immobilized lipase was used for the wax esters synthesis from PSO in a solvent-free system. A conversion of ∼83% was observed after 12 h alcohol/FFA molar ratio 1:1, it increases up to ∼95% with an alcohol/FFA molar ratio of 2:1. After the first three cycles of use, no significant reduction of the lipase activity was observed. Analyses of wax esters properties compared with jojoba oil evidence the feasibility of the produced wax ester for different applications. Iodine number of ∼209 (gI2/100 g oil) was obtained indicating a high degree of anti-oxidant and moisturizing properties.

Lipase‐Catalyzed tert‐Butoxycarbonylation of Alcohols Using Boc2O

AbstractLipase‐catalyzed tert‐butoxycarbonylation of alcohols using di‐tert‐butyl dicarbonate (Boc2O) was developed. Aspergillus niger lipase, Burkholderia cepacia lipase, Pseudomonas fluorescens lipase, Candida rugosa lipase, and Candida antarctica lipase B were examined. The reaction of benzyl alcohol with Boc2O proceeded smoothly in the presence of Candida rugosa lipase in hexane to produce benzyl tert‐butyl carbonate in a good yield. Similarly, the tert‐butoxycarbonylation reactions of 4‐methylbenzyl alcohol, 4‐methoxybenzyl alcohol, 4‐chlorobenzyl alcohol, 4‐bromobenzyl alcohol, 4‐nitrobenzyl alcohol, 2‐phenylethanol, 3‐phenyl‐1‐propanol, cinnamyl alcohol, and 2‐naphthalene methanol using Boc2O in the presence of Candida rugosa lipase in hexane resulted in very good to‐excellent yields of the corresponding tert‐butoxycarbonylated alcohols.

Two phase enzymatic membrane reactor for the production of biodiesel from crude Eruca sativa oil

In the present study, a novel two-phase enzymatic membrane bioreactor (TP-EMR) was developed for the enzymatic biodiesel production from crude Eruca sativa oil. Commercial poly acrylonitrile (PAN) ultrafiltration membrane was examined with a molecular weight cut offs (MWCO) of 30 and 100 kilo Dalton (kDa). Using enzymatic membrane bioreactor with hydrophilic PAN membrane increased the yield of biodiesel production. In this regards, PAN membrane with a MWCO of 100 kDa gave the better results. The inactivation and inhibition effects of methanol and produced glycerol were reduced with the use of membrane bioreactor. It was found that by methanol feeding via aqueous phase, methanol has been penetrated across membrane pores to the organic phase. Afterwards, methanol has reacted with oil at the interface of the membrane at the presence of Candida rugosa lipase (CRL) as a biocatalyst. Produced glycerol, which surrounded the CRL, could be removed from organic phase and conveyed to the aqueous phase to reduce the inhibitory effect of glycerol on the activity of CRL. The yield of about 100% from Eruca sativa oil was achieved in TP-EMR with PAN 100 and organic phase flow rate of 40 ml/min along with 40% initial water content in organic phase.

Efficient lipase-catalysed route for the kinetic resolution of salsolidine and its ß-carboline analogue

Racemic 1-methyl-6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline 1 and 1-methyl-1,2,3,4-tetrahydro-ß-carboline 3 were resolved through lipase-catalysed asymmetric acylation on the secondary amino group. High enantioselectivities (E >200) were observed when the acylation of racemic 1 was performed with phenyl allyl carbonate in the presence of Candida rugosa lipase in toluene at 40 °C or with Candida antarctica lipase B in tert-butyl methyl ether at 50 °C. Excellent enantioselectivity (E >200) characterised the CAL-B-catalysed acylation of racemic 3 with phenyl allyl carbonate in the presence of triethylamine in tert-butyl methyl ether at 50 °C. The product (R)-carbamates (ee >97%) were hydrolysed into the corresponding (R)-enantiomers of the free amines 1 and 3 (ee = 99%) with the use of Pd2(dba)3·CHCl3 catalyst.

ChemInform Abstract: Preparation of Unsymmetrical Dialkyl Acetylenedicarboxylates and Related Esters by Enzymatic Transesterification.

AbstractSymmetrical acetylene dicarboxylates undergo an unexpected highly selective mono‐transesterification with various alcohols in the presence of Candida rugosa lipase.

Preparation of unsymmetrical dialkyl acetylenedicarboxylates and related esters by enzymatic transesterification

An unexpected highly selective mono-transesterification of symmetrical acetylenedicarboxylates with various alcohols occurred in the presence of Candida rugosa lipase. This reaction allows an efficient preparation of unsymmetrical acetylenedicarboxylates and related α,β-acetylenic esters.

Selective biocatalytic aminolysis of (±)-epichlorohydrin: Synthesis and ICAM-1 inhibitory activity of ( S)-(+)-3-arylamino-1-chloropropan-2-ols

Regio- and enantioselective synthesis of ( S)-(+)-3-arylamino-1-chloropropan-2-ols has been achieved by the epoxide ring opening of (±)-epichlorohydrin with different aromatic amines in the presence of Candida rugosa lipase. Activities of seven model ( S)-(+)-3-arylamino-1-chloropropan-2-ols, out of 10 compounds synthesized, have been evaluated for the inhibition of tumor necrosis factor-α TNF-α) induced expression of intercellular adhesion molecule-1 (ICAM-1), which is one of the factors responsible for the modulation of inflammation in biological systems; ( S)-(+)-1-chloro-3-(2′-chlorophenylamino)-propan-2-ol has been found to exhibit highest activity, that is, 86% inhibition of TNF-α induced expression of ICAM-1 at a concentration of 40 μg/ml.

Upgrading of Farmed Salmon Oil Through Lipase-Catalyzed Hydrolysis

Upgrading of farmed salmon oil obtained from by-products was carried out by lipase-catalyzed hydrolysis to increase omega 3 polyunsaturated fatty acids (PUFA) content. The lipases tested were from Penicillium camembertii (PC), Rhizomucor javanicus (RJ), Rhizopus niveus (RN), Rhizopus delemar (RD), Burkholderia cepacia (BC), Rhizopus oryzae (RO), Candida rugosa (CR) and Rhizomucor miehei (RM). The lipases from PC, RJ and RN had lower hydrolysis degrees (HDs) compared to the rest of the lipases. The lipase from CR had the highest HD after 24 h (91.89%). Moreover, CR lipase was the most effective one in concentrating omega 3 PUFA. The final value was increased from 13.77% to 27.81% (wt%). The changes in omega 3 PUFA content were significantly different among the lipases although the HD values were similar at the end of the reactions, which was believed to be caused by the substrate specificities of the lipases. The investigation of the relationship between HD and hydrolysis resistant value (HRV) for eicosapentaenoic acid (EPA, 20:5), docosahexaenoic acid (DHA, 22:6) and oleic acid (OA, 18:1) revealed that the fatty acid (FA) selectivity of the lipases were significantly different. CR lipase had the highest preference for hydrolyzing OA selectively over EPA and DHA. The reaction conditions, i.e. presence of surfactants, sonication, buffer-to-oil ratio, enzyme load, did not affect the selectivity. Investigation of the reaction conditions revealed that it was possible to obtain ~2.15-fold of the original omega 3 PUFA content by hydrolysis of salmon oil in the presence of CR lipase (4%, based on oil weight) with a bufferto- oil ratio of 2:1 (v/v) at 37oC for 4 h.

Optimization of process variables in castor oil hydrolysis by Candida rugosa lipase with buffer as dispersion medium

In this study, castor oil is hydrolyzed in presence of Candida rugosa lipase, while in the buffer (aqueous) phase as a dispersion medium. The following conditions were used to optimize the process: speed of agitation, initial pH of buffer phase, temperature, and ratio of buffer phase volume to oil weight. The optimal conditions are 1,100 rpm, pH 6.5, temperature 35°C, and 3:1 buffer phase volume to oil weight ratio. Under these described conditions, the reusability of lipase was tested and it was found that nearly 80% of original hydrolysis percentage was achieved after the first recycle.

Effect of alkaloids on the activity and selectivity of Candida rugosa lipase in the kinetic resolution of 2-hydroxymethyl-1-phenylthioferrocene with planar chirality

The use of the o-(4-chlorobenzoyl) hydroquinine as an additive in the enzymatic transesterification in the presence of Candida rugosa lipase (CRL) of a planar chiral molecule, such as 2-hydroxymethyl-1-phenylthioferrocene, shows a large enhancement of the reactivity and selectivity of this lipase towards this primary alcohol. An E-value of 143 could be reached at 53% conversion with vinyl acetate as the acylating agent in toluene.

A two steps enzymatic procedure to obtain sterol esters, tocopherols and fatty acid ethyl esters from soybean oil deodorizer distillate

Soybean oil deodorizer distillate (SODD) was enzymatically modified to obtain a product mixture comprised mainly of sterol esters, tocopherols, and fatty acid ethyl esters. Firstly, the original SODD was mixed with oleic acid to reduce its melting point from 65–70 to 30–35 °C and also to produce a reaction mixture with a ratio of free fatty acids (FFA) to sterols close to 2 to improve the progress of sterols esterification. Two enzymatic steps were used in order to separate sterols esterification and ethyl esterification in time and space. The first enzymatic step (in the presence of Candida rugosa lipase) allowed to efficiently transform more than 90% of the original sterols in a short period of time (5 h). The second enzymatic step (in the presence of Novozym 435) converted more than 95% of the FFA in less than 3 h. In addition, the stability of both biocatalysts has been evaluated and both bioprocesses have been scaled-up reutilizing the same batch of lipase up to 8 and 3 times for the first and the second enzymatic step, respectively. The final product obtained is intended to be used as starting material for the purification of sterol esters, tocopherols, and fatty acid ethyl esters via supercritical fluid extraction.

Candida rugosa lipase mediated multigram synthesis of acid part of S(+)-atliprofen, a new NSAID and molecular modeling studies aimed at predicting selectivity of the enzyme

An efficient procedure to prepare S-4-(3-thienyl)phenyl-α-methylacetic acid, an intermediate of a recently approved non-steroidal anti-inflammatory cyclooxygenase inhibitor atliprofen by enantioselective hydrolysis of the corresponding esters in presence of candida rugosa lipase is reported. The methyl and butyl esters of the racemic acid 2 were synthesized and subjected to enantioselective hydrolysis by the lipase to give S-4-(3-thienyl)phenyl-α-methylacetic acid upto 97.86% ee. The observed enantioselectivity during the hydrolysis of the substrate by the lipase was rationalized by molecular modeling studies. The methyl esters of both R and S-enantiomers of 4-(3-thienyl)phenyl-α-methylacetic acid, naproxen and ketoprofen were taken for the modeling studies. The results of the modeling studies are in conformity with the experimental observations.

The importance of ester and alkoxy type functionalities for the chemo- and enantio-recognition of substrates by hydrolysis with Candida rugosa lipase

Racemic esters of 1-phenyl- and 1-(pyridyl)ethyl acetates 1a–c (R = Me, Ph) were subjected to hydrolysis in water in the presence of Candida rugosa lipase (CRL). 1-Pyridylethyl acetates (1, R = Me) are hydrolyzed by crude CRL (C-CRL) and isopropanol (isopropyl alcohol) treated CRL (PT-CRL) at very low rates, and the enantiorecognition was disappointing. By using 1-pyridylethyl benzoates (1, R = Ph) the results differed greatly: hydrolysis occurred much faster, and the enantiorecognition was good for 3- and 4-pyridyl derivatives and excellent for 2-pyridyl derivative. Analogous results were obtained by submitting the 1-phenylethanol esters 4 to enzymatic hydrolysis under the same experimental conditions. The hydrolysis of methyl o-acetoxybenzoates 7 (R = Me) gave quantitatively the deacetylated methyl o-hydroxybenzoates 9 (R = Me) by using either C-CRL or PT-CRL. A complete reversed selectivity is observed in the hydrolysis of phenyl o-acetoxybenzoates 7 (R = Ph) catalyzed by PT-CRL. Molecular modeling studies, aimed at probing the substrate specificity and the enantioselectivity of enzyme in terms of its three-dimensional structure is reported.

Enantioselectivity of alcohol-treated Candidarugosa lipase in the kinetic resolution of racemic methyl 2-aryloxypropionates in water and aqueous organic media

Racemic methyl 2-aryloxypropionates (±)- 1 were subjected to hydrolysis in water and in a series of two-phase aqueous organic media in the presence of Candida rugosa lipase (CRL). The biocatalytic material used was the enzyme of commercial CRL purified by treatment with different alcohols. The purification of CRL and the reaction medium play an important role in the enantioselection of racemates (±)- 1. While it is not possible to use the same protocol for all substrates, by combining the different ways of purifying the enzyme with the various reaction media, it is possible to achieve high enantioselectivities of racemic esters.

Improving the enantioselectivity of Candida rugosa lipase in the kinetic resolution of racemic methyl 2-(2,4-dichlorophenoxy)propionate

Racemic methyl 2-(2,4-dichlorophenoxy)propionate (±)- 1, was subjected to hydrolysis in water and in a series of two-phase aqueous organic media in the presence of Candida rugosa lipase (CRL). The biocatalytic material used was the commercial preparation and enzyme purified by using different procedures. The (+)-R- and (−)-S-2-(2,4-dichlorophenoxy)propionic acids ( 3) were obtained in excellent yield and high enantiomeric excess when the hydrolysis of (±)- 1 was performed in water/benzene in the presence of 2-propanol treated CRL. The kinetic resolution of (±)- 1 was scaled-up.

Enzymatic desymmetrization of meso(anti-anti)-2,4-dimethyl-1,3,5-pentanetriol

Abstract The stereoselective acetylation of meso-3-( tert -butyldimethylsiloxy)-2,4-dimethyl-1,5-pentanediol by vinyl acetate in the presence of Candida rugosa lipase in organic medium gave the corresponding (2R, 3R, 4S) mono ester in high enantiomeric purity (ee = 97%).