Properties Of Candida Rugosa Lipase Research Articles

Improved enzymatic properties of Candida rugosa lipase immobilized on ZnO nanowires/macroporous SiO2 microwave absorbing supports

ZnO nanowires/macroporous SiO2 composites were used as a new type of microwave absorbing support to immobilize Candida rugosa lipase (CRL). Under microwave irradiation the immobilized lipase was used to catalyze esterification of phytosterol and oleic acid for the synthesis of phytosterol esters. A great improvement of the catalytic activity was found in the systems in which composited ZnO nanowires were present. The microwave absorbing properties of ZnO nanowires have been proved to be helpful for the synergistic effect between microwave irradiation and enzyme catalysis. The immobilized CRL showed higher thermal stability at 55°C under microwave irradiation than conventional heating. A maximum molar conversion of 95.4% under microwave irradiation was achieved under the optimal condition: 0.1M phytosterol, 0.2M oleic acid, water activity (aw) of 0.11, reaction temperature at 50°C and reaction time of 1h.

Chemical modification with functionalized ionic liquids: a novel method to improve the enzymatic properties of Candida rugosa lipase

Chemical modification of lysine residues in Candida rugosa lipase (CRL) was carried out using five different functional ionic liquids, and about 15.4-25.0 % of the primary amino groups of lysine were modified. Enzymatic properties of the native and modified CRLs were investigated in olive oil hydrolysis reaction. Improved thermal stability, catalytic activity in organic solvents, and adaptability to temperature and pH changes were achieved compared with the native enzyme. CRL modified by [choline][H2PO4] showed the best results, bearing a maximum improvement of 16.7 % in terms of relative activity, 5.2-fold increase in thermostability (after incubation at 45 °C for 5 h), and 2.3-fold increase in activity in strong polar organic solvent (80 % dimethyl sulfoxide) compared with the native enzyme. The results of ultraviolet, circular dichroism and fluorescence spectroscopy suggested that the change of the secondary and tertiary structures of CRL caused by the chemical modification resulted in the enhancement of enzymatic performance. The modification of CRL with functional ionic liquids was proved to be a novel and efficient method for improving the enzymatic properties of CRL.

Preparation of New Calix[4]arene-Immobilized Biopolymers for Enhancing Catalytic Properties of Candida rugosa Lipase by Sol–Gel Encapsulation

The article describes preparation of new calixarene biopolymers consisting of the immobilization of convenience calixarene derivative onto cellulose and chitosan biopolymers, and the encapsulation of these calixarene biopolymers with Candida rugosa lipase within a chemical inert sol-gel supported by polycondensation with tetraethoxysilane and octyltriethoxysilane. The catalytic properties of immobilized lipase were evaluated into model reactions employing the hydrolysis of p-nitrophenylpalmitate and the enantioselective hydrolysis of naproxen methyl esters from racemic prodrugs in aqueous buffer solution/isooctane reaction system. The resolution studies using sol-gel support have observed more improvement in the enantioselectivity of naproxen E = 300 with Cel-Calix-E than with encapsulated lipase without calixarene-based materials. Furthermore, the encapsulated lipase (Cel-Calix-E) was still retained about 39 % of their conversion ratios after the fifth reuse in the enantioselective reaction.

Effect of ionic liquids, organic solvents and supercritical CO2 pretreatment on the conformation and catalytic properties of Candida rugosa lipase

Abstract The objective of this work is to assess the structure and activity of Candida rugosa lipase (CRL) pretreated with seventeen ionic liquids (ILs), five organic solvents and super-critical carbon dioxide (SC-CO 2 ). The results revealed that anion selection of ILs showed generally much greater effects on CRL esterification activity than cation choice, and CRL pretreated by ILs with strong water miscible properties showed very low esterification activity. The highest CRL activity treated with ILs [Hmim][PF6] was obtained with the value of 45078.0 U/g-protein. Furthermore, the CRL activities pretreated with five conventional organic solvents were also examined and the values increased with the log P decrease of organic solvents when log P was lower than 2.0. Finally, the CRL activities were respectively 1.2- and 1.3-fold higher over the untreated ones after pretreatment with sub- and super-critical CO 2 under the pressures of 6 MPa and 15 MPa at 40 °C for 20 min. Further analyses via FT-IR demonstrated that the high activity of CRL pretreated with ILs, organic solvents and SC-CO 2 was probably caused by the changes of CRL secondary structure. In conclusion, the results in this work will be helpful for us to choose the suitable reaction medium in CRL biocatalysis and biotransformation reactions.

Immobilization of Candida rugosa lipase on poly(3-hydroxybutyrate-co-hydroxyvalerate): a new eco-friendly support

The overall objective of this study is to evaluate the morphological [scanning electron microscopy (SEM)], physicochemical [differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), chemical composition analysis, Fourier-transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR)], and biochemical properties of Candida rugosa lipase (CRL) immobilized on a natural biopolymer poly(3-hydroxybutyrate-co-hydroxyvalerate) (PHBV) in aqueous solution. CRL was immobilized by physical adsorption with efficiency of 30%. Compared with free CRL enzyme, there were slight changes in immobilized CRL activity as a function of temperature (from 37°C to 45°C), but a similar optimal pH value of 7.0. Inactivation rate constants for immobilized CRL enzyme were 0.009 and 0.334 h⁻¹, and half-lives were 77 and 2 h at 40°C and 60°C, respectively. Kinetic parameters obtained for immobilized CRL include the Michaelis-Menten constant of K(m) = 213.18 mM and maximum reaction velocity of V(max) = 318.62 U/g. The operational stability of immobilized CRL was tested repeatedly, and after 12 cycles of reuse, the enzyme retained 50% activity. Based on our results, we propose that PHBV-immobilized CRL could serve as a promising biocatalyst in several industrial applications.

Improvement of catalytic properties of Candida Rugosa lipase by sol–gel encapsulation in the presence of magnetic calix[4]arene nanoparticles

Candida rugosa lipase (CRL) was encapsulated within a chemically inert sol-gel support prepared by polycondensation with tetraethoxysilane (TEOS) and octyltriethoxysilane (OTES) in the presence of N-methylglucamine based calix[4]arene magnetic nanoparticles. The results indicate that the magnetic calix[4]arene based encapsulated lipase particularly has shown high conversion and enantioselectivity. It has also been noticed that the magnetic calix[4]arene based encapsulated lipase has excellent enantioselectivity (E = 460) as compared to the free enzyme (E = 166) with an ee value of >98% for S-Naproxen.

Influence of the functional activating agent on the biochemical and kinetic properties of Candida rugosa lipase immobilized on chemically modified cellulignin

Candida rugosa lipase was immobilized by covalent binding on wood cellulignin ( Eucaliptus grandis) chemically modified with different activating agents as carbonyldiimidazole, glutaraldehyde and sodium metaperiodate. The resulted immobilized derivatives were evaluated in both aqueous (hydrolysis) and organic (ester synthesis) media. In aqueous media a comparative study between free and immobilized derivatives was provided in terms of pH, temperature and kinetic constants ( V max and K m) following the hydrolysis of p-nitrophenyl palmitate, in which new optima values were established. The experimental results suggested that functional activating agents render different interactions between enzyme and cellulignin, producing consequently alterations in the optimal reaction conditions. Different behavior was found when the immobilized derivatives were tested in organic media, under these conditions similar esterification activities were observed, independent on the agent used to active the immobilizing support. Reasons for this are discussed on the light of the interactions among the support, functional activating agent and lipase structure.