Support For Enzyme Immobilization Research Articles

Lipase B from Candida antarctica Immobilized on Epoxy-functionalized Hollow Silica Microspheres: Efficient Biocatalysts for Enantiomer Selective Acylation of Alcohols and Amines

Hollow silica microspheres with promising physical properties (MAT540TM) as support for enzyme immobilization and biocatalyst were investigated in this study. The amine-functionalized MAT540TM was activated by six bisepoxides inclosing different spacers and used as epoxy-functionalized carrier for immobilization of lipase B from Candida antarctica (CaLB). The novel, covalently fixed CaLB biocatalysts were compared in kinetic resolution (KR) of racemic 1-phenyethanol rac-1 and five racemic amines rac-3a-e using shaken flasks and continuous-flow packed-bed microreactors. Mechanic stability, re-usability and the effect of temperature (0–90 °C) on productivity and enantiomer selectivity of the covalently immobilized CaLB were investigated. The best performing CaLB biocatalyst showed good mechanic stability after 24 h operation time in continuous-flow mode at 60 °C and provided in KRs of racemic 1-phenyethanol rac-1 with vinyl acetate and of five racemic amines with isopropyl 2-ethoxyacetate as acylating agent the non-reacted (S)-alcohol [(S)-1] or (S)-amines [(S)-3a-e] and the forming (R)-ester [(R)-2] or (R)-amide [(R)-4a-e] in good yields with high enantiomeric excess (ee > 99 %, for all).

Polymer Nanofibrous Material for Enzyme Immobilization

NanospiderTMmethod allows preparation of nanofiber sheets with desired specific mass and fiber diameter between 100-600 nm. The fiber diameter can be modified by solvent system, concentration of polymer solution, and external conditions of the electrospinning process, in particular by air humidity and temperature. We have long experience with continual electrospinning from free surface of the polymer solution and in this case we focus on polymer blend polyamide 6/chitosan nanofiber form. Chitosan is an excellent material providing biocompatibility and numerous primary amine groups. On the other hand, polyamide 6 is well known and easy-to-prepare stable polymer guaranteeing sufficient mechanical properties. Blend nanofibers containing synthetic and natural polymers are usually difficult to continuously prepare. Natural polymer bring certain inhomogeneity due to varying chemical and physical properties of each brew. We compared chitosan from four different suppliers and finally fabricated homogenous and long-term stable layers. Successfully prepared nanofiber sheets were used as a support for enzyme immobilization. We showed that our layers can be used as a carrier for numerous biomolecules.

A novel amperometric bienzymatic biosensor based on alcohol oxidase coupled PVC reaction cell and nanomaterials modified working electrode for rapid quantification of alcohol

A new amperometric sensor has been fabricated for sensitive and rapid quantification of ethanol. The biosensor assembly was prepared by covalently immobilizing alcohol oxidase (AOX) from Pichia pastoris onto chemically modified surface of polyvinylchloride (PVC) beaker with glutaraldehyde as a coupling agent followed by immobilization of horseradish peroxidase (HRP), silver nanoparticles (AgNPs), chitosan (CHIT), carboxylated multi-walled carbon nanotubes (c-MWCNTs) and nafion (Nf) nanocomposite onto the surface of Au electrode (working electrode). Owing to properties such as chemical inertness, light weight, weather resistance, corrosion resistance, toughness and cost-effectiveness, PVC membrane has attracted a growing interest as a support for enzyme immobilization in the development of biosensors. The amperometric biosensor displayed optimum response within 8 s at pH 7.5 and 35°C temperature. A linear response to alcohol in the range of 0.01mM–50 mM and 0.0001 µM as a minimum limit of detection was displayed by the proposed biosensor with excellent storage stability (190 days) at 4°C. The sensitivity of the sensor was found to be 155 µA mM−1 cm−2. A good correlation (R2 = 0.99) was found between alcohol level in commercial samples as evaluated by standard ethanol assay kit and the current biosensor which validates its performance.

Synergistic Degradation of Dye Wastewaters Using Binary or Ternary Oxide Systems with Immobilized Laccase

In recent years, groundwater contamination caused by dyes has become an important problem. They enter into wastewater as a result of the textile, automotive, or cosmetics industries. For this reason, new methods are being sought, which would aid at the removal of dye impurities with high efficiency and also would be relatively cheap. In the presented study synthesized TiO2-ZrO2 (with TiO2:ZrO2 molar ratio of 8:2) and TiO2-ZrO2-SiO2 (with TiO2:ZrO2:SiO2 molar ratio of 8:1:1) oxide materials were used as supports for enzyme immobilization. Effective synthesis of the carriers was confirmed by results of scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), low-temperature nitrogen sorption and Fourier transform infrared spectroscopy (FTIR). The materials achieve high immobilization efficiency of the laccases from Trametes versicolor (83% and 96% for TiO2-ZrO2-laccase and TiO2-ZrO2-SiO2-laccase, respectively). The effect of selected dye concentrations, pH, temperature, and reusability were also tested. The obtained results showed that after removal of textile dyes, such as Alizarin Red S (ARS), Remazol Brilliant Blue R (RBBR), and Reactive Black 5 (RB5), under optimal process conditions, which were pH 5 and 25 °C, from dye solution of 5 mg/L degradation efficiency reached 100%, 91%, and 77%, respectively, suggesting synergistic mechanism of degradation by simultaneous sorption and catalytic action. Finally, reduction of chemical oxygen demand (COD) of the solution after treatment indicated lower mixture toxicity and effective dye degradation.

Palm oil fatty acids and carotenoids extraction with lipase immobilized in magnetic nanoparticles

Magnetite nanoparticles have unique properties including superparamagnetism and low toxicity. They have been used as supports for immobilization of enzymes because of the advantage including easy separation and effective recycle under external magnetic field. Therefore, the present study aimed at developing a new enzymatic biocatalyst from the immobilization of Yarrowia lipolytica’s (IMUFRJ 50682) lipase on magnetic nanoparticles for its use in free fatty acids and carotenoids extraction from palm oil (Elaeis guineenses). The lipase production conducted in a 4 L benchtop bioreactor generated a crude enzyme extract with hydrolytic p-nitrophenyl laurate activity of 58 U/ml after 24 h. Lipase was immobilized by adsorption on magnetite nanoparticles (Fe3O4). The crude enzymatic extract, both free and immobilized, were used in the hydrolysis of palm oil. Temperature, reaction time and substrate ratio (water/palm oil) were evaluated in a central composite experimental design. The initial concentration of fatty acids and carotenoids present in palm oil obtained via traditional extraction (using organic solvents) were 82.45% and 1892.3 mg/kg, respectively. Using crude lipase extract immobilized in nanoparticles, after 120 min, temperature of 24°C and water/oil ratio of 2, it was possible to obtain similar values, with the advantage of easily removing the catalyst from reaction media and reusing it.

Biocompatible magnetic nanoparticles grafted by poly(carboxybetaine acrylamide) for enzyme immobilization

Herein, the zwitterionic material poly (carboxybetaine acrylamide) was grafted onto iron oxide to obtain biocompatible magnetic nanoparticles Fe3O4-pCBAA which were employed to immobilize enzymes. The nanocomplxes Fe3O4-pCBAA were characterized using scanning electron microscopy (SEM), dynamic light scattering (DLS), zeta potential, Fourier transform-infrared (FT-IR) spectra and energy dispersive X-ray spectrometry (EDX). The urease as a model enzyme was immobilized with the novel supports and the properties of immobilized urease were further investigated in comparison with the free urease counterpart. The immobilized urease exhibited excellent thermodynamic and chemical stability. Particularly, 60% of initial activity was remained after being stored at 70 °C for 2 h while the free urease only remained 30%. Besides, the relative activity of immobilized enzyme was 1.7 times that of free ones after disposed in ethanol and 2-propanol for 2 h, and 7 times in DMF. Moreover, immobilized urease retained >80% of its initial activity after 5 cycles. In addition, the immobilization carrier Fe3O4-pCBAA displayed famous biocompatibility, and the immobilized urease performed better in complex biological samples, which were >85% and <60% of its initial activity for the immobilized and dissociative urease, respectively, in 20% and 25% of serum. These results confirm that the nanoparticles Fe3O4-pCBAA are biofriendly and efficient supports for enzyme immobilization and potential for practical applications in bio-microenvironments.

Fabrication of AChE/SnO2-cMWCNTs/Cu Nanocomposite-Based Sensor Electrode for Detection of Methyl Parathion in Water

The work highlights inhibition-based Acetylcholinesterase (AChE) fabrication using composite nanomaterial comprising tin oxide nanoparticles (SnO2) and carboxylated multiwalled carbon nanotubes (cMWCNTs) for detection of pesticide methyl parathion (MP) in water samples. Working electrode AChE/SnO2-cMWCNTs/Cu exhibited high sensitivity with a linearity range of 1.0 μM to 160 μM and a minimum detection limit of 0.1 μM for MP in water. The fabricated electrode was found biocompatible and nontoxic which can be used to detect low concentrations of pesticide in water samples. The synergistic and facilitated electron transferring properties of SnO2-cMWCNTs/Cu made it an excellent support for immobilization of enzyme in sensing technology. The enzyme AChE was covalently immobilized with cMWCNTs using glutaraldehyde as crosslinking agent which has enhanced the storage stability and reusability of the method. The reusability attained was 30 times for 40 days when AChE/SnO2-cMWCNTs/Cu was stored at low temperature of 4°C. Developed sensor showed excellent analytical recovery of pesticide in water sample with negligible effect of interfering species. Also, AChE/SnO2-cMWCNTs/Cu was easily reactivated simply by varying pH of phosphate buffer. This method is fast, reliable, and accurate showing successful development of amperometric biosensor for detection of MP in water sample.

Synthesis of Chitosan-Functionalized Fibrous Membrane for Immobilization of Horseradish Peroxidase: Interfacial Property and Application for Catalytic Oxidation of P-Nitrophenol

A hydrophilic fibrous membrane consisting of a network of a non-crystalline hydrophobic polymer and an in situ crosslinked hydrophilic chitosan polymer is studied. This chitosan-functionalized membrane can be employed as a support for immobilization of horseradish peroxidases (HRPs). The immobilized HRPs (mass loading ~110 mg /g) show high stabilities over a wide pH range of 4~10 and temperature range of 25~60°C. In addition, the immobilized HRPs can function as a recyclable biocatalyst for catalytic oxidation of p-nitrophenol (PNP). After 10 cycles, the PNP removal efficiency is 70 percent. The results demonstrate that the chitosan-functionalized fibrous membrane is a promising support for immobilization of enzymes and has applications related to enzyme catalysis.

Effective Enzyme Immobilization onto a Magnetic Chitin Nanofiber Composite

In this study, a novel biocompatible magnetic chitin nanofiber composite (MCNC) was developed as a support for enzyme immobilization, and the enzyme-immobilizing ability was elucidated using chymotrypsin (CT) as a model enzyme. Chitin nanofibers (CNFs) were prepared via 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-mediated oxidation of chitin and then further modified with magnetic nanoparticles. Glutaraldehyde was used to cross-link the additional CT molecules and aggregate them onto the MCNCs. The CNFs were characterized by transmission electron microscopy, and Fourier transform infrared spectroscopy. The results showed that the CNFs were properly formed and that the CT molecules immobilized on the MCNCs presented excellent properties. After heating the composites at 60 °C for 3 h, the non-cross-linked and cross-linked immobilized CTs retained 51.6% and 70.7% of the initial activity, respectively, whereas the free CTs retained only 29.6% of the initial activity. In addition, non-cross-linked and cross-l...

Laccase Immobilization onto Magnetic β-Cyclodextrin-Modified Chitosan: Improved Enzyme Stability and Efficient Performance for Phenolic Compounds Elimination

Three types of improved Fe3O4 magnetic nanoparticles (MNPs), including poly(amidoisophthalicacid) coated magnetite nanoparticles (Fe@PA), cyclodextrin (CD) anchored Fe@PA (Fe@PA-CD), and chitosan (Cs) coated Fe@PA-CD (Fe@PACD-Cs) were successfully developed and characterized. Laccase immobilization onto MNPs was carried out via physical adsorption. The maximal and minimal loading capacity were obtained for Fe@PA and Fe@PA-CD-Cs, respectively. Fe@PA-CDCs-laccase exhibited around 100% of the maximum activity at pH 4 and maintained 70% of its initial activity within the temperature range of 15–55 °C; and Cs coated nanoparticles were more efficient than non-coated. Fe@PA-CD-Cs-laccase maintained 70% of its initial activity up to 12 d from the first day of storage at 25 °C whereas the free laccase, Fe@PA-laccase, and Fe@PA-CD-laccase kept 10%, 28%, and 33% of initial activity, respectively. Furthermore, bio-removal of phenolic compounds was performed by the free and immobilized enzyme. Fe@PA-CD-Cs-laccase showed maximal removal with 96.4% and 85.5% for phenol and bisphenol A, respectively. It seems that Fe@PA-CD-Cs could be an appropriate support for immobilization of other enzymes in various industrial application especially bioremoval of phenolic compounds.

Immobilization of the enzyme invertase in SBA-15 with surfaces functionalized by different organic compounds

The search for appropriate supports for enzyme immobilization has attracted increasing interest in the field of biocatalysis. Immobilized enzymes are catalysts of enormous industrial interest because they combine the advantages of heterogeneous catalysis with the high selectivity and mild operational conditions of enzymes. This study evaluated the use of different organic molecules as functionalizing agents anchored on the silanol groups present on the surface of mesoporous materials such as SBA-15. This modification aims to increase the interaction of the enzyme invertase to the support, allowing for a greater amount of this biocatalyst to be immobilized. The hexagonal mesoporous material SBA-15, which has pores in the range of 40–60 A, was synthesized and modified by the following organic compounds: trimethoxyphenylsilane, vinyltrimethoxysilane and 3-aminopropyltriethoxysilane (APTES). APTES showed the best results in increasing the anchoring of enzymes on the material’s surface, providing a specific amount of enzyme loaded onto the surface of approximately 80% and, thus, showing successful functionalization.

Development of laccase/graphene oxide membrane for enhanced synthetic dyes separation and degradation

Enzymatic degradation with laccase is a green option for removal of organic compounds from aqueous media. However, the development of high-performance support for immobilization of enzyme is constantly required for the advancement of this technology. In this study the laccase was facilely immobilized on graphene oxide (GO) nanosheets separation membrane, and the membrane’s performance as laccase immobilization carrier has been investigated. The results showed that the immobilized laccase on GO-PES membrane exhibited excellent stability towards temperature and pH in the long term and the flux of pure water of biocatalytic membrane was enhanced as well. Synthetic dye degradation was carried out with four representative dyes with different molecular size and structure. The biocatalytic membrane showed substantial improvement of these dyes degradation compared with the GO-PES membrane without laccase immobilized. The relative activity order of the immobilized laccase for the degradation of these dyes was Reactive Brilliant Blue (K-GR) > Reactive Brilliant Blue (KN-R) > Methyl Orange (MO) > Crystal Violet (CV). This order was discussed in terms of correlation between molecular size and structure of dye species. GO coating membranes with catalytic property of laccase offer a new potential approach towards synthetic dye degradation.

Pinewood nanobiochar: A unique carrier for the immobilization of crude laccase by covalent bonding

Nanotechnology-inspired biocatalytic systems attracted attention for many applications since nanosized supports for enzyme immobilization can improve efficiency-determining factors e.g. enhancing the surface area and loading capacity and reducing the mass transfer resistance. Among the nanomaterials, nanobiochar has unique features as a support for enzyme immobilization i.e. high surface to volume ratio, porous structure, and presence of functional groups on its surface. However, the performance of the immobilization is highly dependent on the immobilization conditions and the properties of the enzyme and the support material. In this research, crude laccase was covalently immobilized onto functionalized nanobiochar using a two-step method of diimide-activated amidation. The effect of different parameters was investigated. The optimal conditions were found to be 14 mg/mL of laccase concentration, 5 mg/mL of nanobiochar, 8.2 mM of cross-linker and 3 h of contact time. For investigating the pH, thermal, storage, and operational stability, the sample obtained from the optimized conditions was used. The results showed the higher stability of immobilized laccase against temperature and pH variation compared to free laccase. In addition, immobilized laccase maintained its catalytic performance up to seven cycles of utilization and showed more than 50% of initial activity after two months of room temperature storage.

Preparation of Polydopamine-Modified 3D Interconnected Macroporous Silica for Laccase Immobilization

Millimeter-sized polydopamine (PDA) modified three-dimensional interconnected macroporous silica (PDA/3DIMS) with high porosity, large pore volume and pore diameter was prepared and used as the support to immobilize commercial laccase (Lac) from Denilite II S. The skeleton of 3DIMS is constructed by silica nanofilm, so the PDA/3DIMS is constructed by PDA/SiO2/PDA sandwich nano-film. The immobilization conditions, incubation time, pH and enzyme concentration were optimized, and the properties of the immobilized laccase were investigated. The enzyme activity of Lac-PDA/3DIMS (295.9 U/g) was higher than that of Lac-3DIMS (222.2 U/g). The stability and reusability of the two immobilized laccases were both improved comparing with the free laccase. Lac-PDA/3DIMS exhibited higher retained activity (73.6%) than Lac-3DIMS (31.8%) after reused for ten times. Lac-PDA/3DIMS retained more than 84.0% initial activity after storage for a month, however, the free laccase and Lac-3DIMS were 7.1% and 48.6%, separately. These advantages revealed that PDA thin layer plays an important role to improve the enzymatic activity of immobilized laccase, and the PDA/3DIMS material is a very promising support for enzyme immobilization.

Effect of enzymatic hydrolysis on developing support of polyamide woven fabric for enzyme immobilization

Polyamide fiber has been considered as a suitable support for enzyme immobilization because of its low cost, chemical and mechanical properties and ready availability in a number of different forms. In particular, polyamide fabric has highly specific surface and good elasticity. The present study aims to develop an immobilization support from polyamide fabric and to establish the optimum immobilization conditions for laccase. For this, the enzymatic hydrolysis process was introduced to the hydrolysis of polyamide, creating amino groups that immobilize enzyme molecules. When polyamide fabric was hydrolyzed by bromelain during enzymatic hydrolysis, the highest immobilization yield (68 ± 0.7%) and relative activity (95 ± 0.52%) of immobilized laccase were obtained. For successful enzyme immobilization, the optimal glutaraldehyde crosslinking conditions were pH of 9.0 with 10% (v/v) of glutaraldehyde concentration for 240 min at 45℃. The most favorable immobilization conditions were as follows: pH of 6.0 with 35% (owf) of laccase concentration for 600 min at 4℃. Under the optimum treatment conditions, the pH and thermal stability of immobilized laccase were improved. After 20 days of storage, the immobilized laccase on enzymatic hydrolyzed polyamide fabric retained approximately 30% of its initial activity. Furthermore, the immobilized laccase indicated potential reuse over 10 use cycles. The structural changes of polyamide fabric according to treatment processes were demonstrated by Fourier transform infrared spectroscopy. The changes of surface morphology were measured by scanning electron microscopy according to the multi treatment steps.

An ideal enzyme immobilization carrier: a hierarchically porous cellulose monolith fabricated by phase separation method

Abstract Cellulose monolith with a hierarchically porous morphology was utilized as a novel solid support for enzyme immobilization. After a series of modifications, succinimidyl carbonate (SC)-activated cellulose monolith (SCCL monolith) was obtained and it was employed to immobilize a model enzyme (horseradish peroxidase, HRP) through covalent bonding. The HRP immobilization capacity on SCCL monolith was calculated as 21.0 mg/g. The thermal stability measurement illustrated that the immobilized HRP exhibited a largely improved thermal resistance compared to its free counterpart. The reusability of the immobilized HRP was investigated, and it could be reused at least 10 cycles without significant activity loss. Therefore, cellulose monolith is found to be an ideal solid support for enzyme immobilization.

EFFECT OF REACTION CONDITIONS ON THE SYNTHESIS OF CYCLODEXTRIN (CD) BY USING IMMOBILIZED ENZYME

The production of cyclodextrin (CD) over the years has been increasing due to the numerous applications in industries such as in food, cosmetic, pharmaceutical and agricultural industries. However, cyclodextrin glucanotransferase (CGTase) which involved in the enzymatic reaction on the production of CD is unstable and easily denatured at extreme conditions resulted in low CD production. Hence, the enzyme immobilization technique is introduced to overcome these problems and subsequently increase the production of CD. In the present study, the CGTase was immobilized on hollow fiber membrane to increase the production of CD during the reaction. The effect of reaction conditions (types of starch, concentration of starch, temperature and pH) of the immobilized enzyme on the production of CD were investigated. Among the three types of starch tested, the soluble potato starch was the most suitable substrate for the production of CD with 4.13 mg/mL. In addition, by using 3% (w/v) of the soluble potato starch, the production of CD was 5.22 mg/mL . The optimal reaction temperature and pH were found to be at 40°C and pH 6 with 5.21 mg/mL and 4.62 mg/ml of CD, respectively. The immobilized enzyme exhibited a 1.3-3-fold increase in CD production compared to the free enzyme. Therefore, the hollow fiber membrane is suitable to be used as a support for enzyme immobilization with the high production of CD.

Biogenic porous silica and silicon sourced from Mexican Giant Horsetail (Equisetum myriochaetum) and their application as supports for enzyme immobilization.

Porous silica-based materials are attractive for biomedical applications due to their biocompatibility and biodegradable character. In addition, inorganic supports such as porous silicon are being developed due to integrated circuit chip compatibility and tunable properties leading to a wide range of multidisciplinary applications. In this contribution, biosilica extracted from a rarely studied plant material (Equisetum Myriochaetum), its conversion to silicon and the potential for both materials to be used as supports for enzyme immobilization are investigated. E. myriochaetum was subject to conventional acid digestion to extract biogenic silica with a% yield remarkably higher (up to 3 times) than for other Equisetum sp. (i.e. E. Arvense). The surface area of the isolated silica was ∼400 m2/g, suitable for biotechnological applications. Biogenic silicon was obtained by magnesiothermic reduction. The materials were characterized by SEM-EDX, XRD, FT-IR, ICP-OES, TGA and BET analysis and did not contain significant levels of class 1 heavy elements (such as Pb, Cd, Hg and As). Two commercial peroxidases, horseradish peroxidase (HRP) and Coprinus cinereus peroxidase (CiP) were immobilized onto the biogenic materials using three different functionalization routes: (A) carbodiimide, (B) amine + glutaraldehyde and (C) amine + carbodiimide. Although both biogenic silica and porous silicon could be used as supports differences in behaviour were observed for the two enzymes. For HRP, loading onto biogenic silica via the glutaraldehyde immobilization technique (route B) was most effective. The loading of CiP showed a much higher peroxidase activity onto porous silicon than silica functionalized by the carbodiimide method (route A). From the properties of the extracted materials obtained from Equisetum Myriochaetum and the immobilization results observed, these materials appear to be promising for industrial and biomedical applications.

Preparation and characterization of alkaline phosphatase, horseradish peroxidase, and glucose oxidase conjugates with carboxylated carbon nano-onions

ABSTRACTCarbon nanomaterials have emerged as suitable supports for enzyme immobilization and stabilization due to their inherently large surface area, high electrical conductivity, chemical stability, and mechanical strength. In this paper, carbon nano-onions (CNOs) were used as supports to immobilize alkaline phosphatase, horseradish peroxidase, and glucose oxidase. CNOs were first functionalized by oxidation to generate carboxylic groups on the surface followed by the covalent linking of using a soluble carbodiimide as coupling agent. The CNO–enzyme conjugates were characterized by transmission electron microscopy and Raman spectroscopy. Thermogravimetric analysis revealed a specific enzyme load of ∼0.5 mg of protein per milligram of CNO. The immobilized enzymes showed enhanced storage stability without altering the optimum pH and temperatures. These properties make the prepared nanobiocatalyst of potential interest in biosensing and other biotechnological applications.

Preparation and Characterization of Cellulose Triacetate as Support for Lecitase Ultra Immobilization.

The use of polymers as supports for enzyme immobilization is a strategy that enables to remove the enzymes from a chemical reaction and improve their efficiency in catalytic processes. In this work, cellulose triacetate (CTA) was used for physical adsorption of phospholipase Lecitase ultra (LU). CTA is more hydrophobic than cellulose, shows good performance in the lipases immobilization being a good candidate for immobilization of phospholipases. We investigated the immobilization of LU in CTA, the stability of the immobilized enzyme (CTA-LU) and the performance of CTA-LU using soybean oil as a substrate. LU was efficiently immobilized in CTA reaching 97.1% in 60 min of contact with an enzymatic activity of 975.8 U·g−1. The CTA-LU system presents good thermal stability, being superior of the free enzyme and increase of the catalytic activity in the whole range of pH values. The difference observed for immobilized enzyme compared to free one occurs because of the interaction between the enzyme and the polymer, which stabilizes the enzyme. The CTA-LU system was used in the transesterification of soybean oil with methanol, with the production of fatty acid methyl esters. The results showed that CTA-LU is a promising system for enzymatic reactions.