Enzyme immobilisation technology is an effective means to improve sugar ester production through the employment of biocatalysts. In the present study, immobilisation of Candida rugosa (CRL) lipase onto amino-activated mica is performed via covalent bonding (namely Amino-CRL) and the cross-linking of lipases into nano-reactors through physical adsorption (namely NER-CRL). Free and immobilised lipases were tested for their esterification activities. Specific activities for Amino-CRL and NER-CRL increased by 2.4 and 2.6-fold, respectively, upon immobilisation. Extending this work, immobilised lipases have novel capabilities in the synthesis of sugar esters. The optimised conditions for sugar fatty acid ester syntheses are 48 h at 2:1 of molar ratio of lactose sugar to capric acid at 55 �C. Furthermore, a high operational stability with half-lives of over 13 and 10 runs was achieved for NER-CRL and Amino-CRL, respectively, indicating the efficiency of the immobilisation process. � 2011 Elsevier Ltd. All rights reserved.

The utilization of natural mica as a biocatalyst support in kinetic investigations is first described in this study. The formation of lactose caprate from lactose sugar and capric acid, using free lipase (free-CRL) and lipase immobilized on nanoporous mica (NER-CRL) as a biocatalyst, was evaluated through a kinetic study. The apparent kinetic parameters, Km and Vmax, were determined by means of the Michaelis-Menten kinetic model. The Ping-Pong Bi-Bi mechanism with single substrate inhibition was adopted as it best explains the experimental findings. The kinetic results show lower Km values with NER-CRL than with free-CRL, indicating the higher affinity of NER-CRL towards both substrates at the maximum reaction velocity (Vmax,app > Vmax). The kinetic parameters deduced from this model were used to simulate reaction rate data which were in close agreement with the experimental values.

The kinetics of wax ester synthesis from oleic acid and oleyl alcohol using immobilized lipase from Candida antartica as catalyst was studied with different types of impeller (Rushton turbine and AL-hydrofoil) to create different mixing conditions in 2l stirred tank reactor. The effects of catalyst concentration, reaction temperature, and impeller tip speed on the synthesis were also evaluated. Rushton turbine impeller exhibited highest conversion rate at lower impeller tip speed as compared to AL-hydrofoil impeller. A second-order reversible kinetic model from single progress curve for the prediction of fractional conversion at given reaction time was proposed and the corresponding kinetic parameter values were calculated by non-linear regression method. The results from the simulation using the proposed model showed satisfactory agreement with the experimental data. Activation energy shows a value of 21.77 Kcal/mol. The thermodynamic parameters of the process, enthalpy and entropy, were 21.15 Kcal/mol and 52.07 cal/mol.K, respectively. � 2010 by Japan Oil Chemists' Society.

Amino acid surfactants are high-value surfactants which have excellent emulsifying characteristics and minimal toxicity to the living body. Enzymatic synthesis of palm kernel amino acid surfactant was optimized by response surface methodology (RSM) using palm kernel olein (PKO) and l(+)-lysine catalyzed by Lipozyme RM IM. The reaction was performed in batch mode stirred tank reactor (STR) with one multi-bladed impeller. A central composite rotatable design (CCRD) was employed to evaluate the interactive effects of various parameters. The parameters were temperature (A): (40.00-70.00 �C), impeller speed (B): (100.00-400.00 rpm), substrates ratio (C): (1.00-4.00 mmol) and enzyme amount (D): (5.00-8.00 g). The optimum condition derived via RSM at fixed reaction time of 24 h was temperature; 47.50 �C, impeller speed; 323.96 rpm, substrates ratio; 3.25 mmol and enzyme amount; 7.25 g. The experimental yield was 89.03% under the optimum condition, which compared well with the maximum predicted value of 93.77%. � 2009 Elsevier B.V. All rights reserved.

Optically active (-)-menthyl butyrate was synthesized by enantioselective esterification of racemic (±)-menthol and butyric anhydride using lipase from Candida rugosa immobilized onto epoxy-activated supports of Eupergit C and Eupergit C 250 L through physical adsorption method. The effects of various temperature, storage condition, stability in organic solvent and lipase recyclability were investigated for their influence on the enzymatic enantioselective formation of (-)-menthyl butyrate. The immobilized lipases retained high catalytic activity of up to 31% yield and 100% enantiomeric excess of the desired product, and showed better stability compared to the native lipase. They also exhibited about 50% retained activity even after incubation at higher temperatures, storage at room temperature and after long incubation in hexane. Immobilized lipases also showed considerably efficient reusability.

Mica was modified either by acid treatment, grafting with aminopropyl-, octyl-, vinyl-, mercapto- and glycidoxy-triethoxysilanes, and activation of pre-treated support with glutaraldehyde (Glu). The derivatives were characterized by X-ray diffraction (XRD), infra-red spectroscopy (FTIR), surface area and porosity analysis, scanning electron microscopy coupled with energy dispersive X-ray (SEM-EDX) and transmission electron microscopy (TEM) techniques. The modified micas were used for immobilization of lipase from Candida rugosa (CRL). Activity of the lipase was determined by esterification and exhibited the improved activity than the free enzyme following the order; Amino-CRL > Glu-Amino-CRL > Octyl-CRL > Vinyl-CRL > Glycidoxy-CRL > Mercapto-CRL > Mica-CRL. Lipase immobilized mica showed enhanced protein loading (up to 8.22 mg protein/g support) and immobilization (up to 78%) compared to the free lipase and unmodified mica. � 2009 Elsevier B.V. All rights reserved.