Carrier For Immobilization Research Articles

Enhancing Osmotic Energy Harvesting Through Supramolecular Design of Oxygen‐Functionalized MXene with Biomimetic Ion Channels

AbstractReverse electrodialysis (RED) technology, relying on ion‐selective permeability membranes (ISPM), offers a direct means of harnessing osmotic energy from the salinity difference between seawater and freshwater. Critical technical challenges include limitations in ISPM's immobile charge carriers, transmembrane ionic internal resistance, and durability in water. Drawing inspiration from the subunit structure of the epithelial sodium channel (ENaC), an ISPM assembled using a supramolecular engineering strategy is introduced. This innovative approach enables the synergistic action of multiple molecular building blocks to mimic the distribution of ENaC subunit structures, strategically planning the placement of immobile charge carriers on nanofluidic channels. The design incorporates nano‐confined oxygen‐rich functionalized MXene (O‐MXene) and high‐strength polymer backbone aramid fibers to construct 3D nanofluidic channels with a high surface charge density. Enhanced by a bonding network of sodium carboxymethylcellulose, the ISPM achieved an exceptional output power density of 21.7 W m−2 and 46.0% energy conversion efficiency in a RED half‐cell system using natural seawater and river water, surpassing current technologies. This research not only advances RED technology but also provides biomimetic customization concepts for ISPM design across various applications, including flow batteries, fuel cells, and related fields.

Hydroxyapatite: An Eco-Friendly Material for Enzyme Immobilization.

Biocatalysis has emerged in the last decade as a valuable and eco-friendly tool in chemical synthesis, allowing in several instances to reduce or eliminate the use of hazardous reagents, environmentally dangerous solvents and harsh reaction conditions. Enzymes are indeed able to catalyse chemical transformations on non-natural substrates under mild reaction conditions, still maintaining their high chemo-, regio-, and stereoselectivity. Enzyme immobilization, i. e. the grafting of enzymes on solid supports, can be viewed as an enabling technology, as it allows a better control of the reaction and the recycling of the biocatalyst, thus rendering economically viable the use of expensive enzymes also on a large scale. To pursue a sustainable approach, the supports for enzyme immobilization should be eco-friendly and possibly renewable. This review highlights the use of hydroxyapatite (HAP), an inorganic biomaterial able to confer strength and stiffness to the bone tissue in animals, as carrier for enzyme immobilization. HAP is a cheap, non-toxic and biocompatible material, with high surface area and protein affinity. Different enzyme classes, immobilization strategies, and the use of diverse HAP-based supports will be discussed, underlining the immobilization conditions and the properties of the obtained biocatalysts.

Nanodiamonds and natural deep eutectic solvents as potential carriers for lipase

This study investigates the use of nanodiamonds (ND) as a promising carrier for enzyme immobilization and compares the effectiveness of immobilized and native enzymes. Three different enzyme types were tested, of which Rhizopus niveus lipase (RNL) exhibited the highest relative activity, up to 350 %. Under optimized conditions (1 h, pH 7.0, 40 °C), the immobilized ND-RNL showed a maximum specific activity of 0.765 U mg−1, significantly higher than native RNL (0.505 U mg−1). This study highlights a notable enhancement in immobilized lipase; furthermore, the enzyme can be recycled in the presence of a natural deep eutectic solvent (NADES), retaining 76 % of its initial activity. This aids in preserving the native conformation of the protein throughout the reusability process. A test on brine shrimp revealed that even at low concentrations, ND-RNL had minimal toxicity, indicating its low cytotoxicity. The in silico molecular dynamics simulations performed in this study offer valuable insights into the mechanism of interactions between RNL and ND, demonstrating that RNL immobilization onto NDs enhances its efficiency and stability. All told, these findings highlight the immense potential of ND-immobilized RNL as an excellent candidate for biological applications and showcase the promise of further research in this field.

Multivariate mesoporous MOFs with regulatable hydrophilic/hydrophobic surfaces as a versatile platform for enzyme immobilization

Zeolitic imidazole frameworks materials-8 (ZIF-8), a member of the metal-organic framework (MOFs) series, have been extensively used as a host matrix for enzyme immobilization. However, the microporous structure and hydrophobicity, as well as the protonation of the precursor 2-methylimidazole (2-MeIm) of ZIF-8, remain considerable challenges in maintaining the activity of immobilized enzymes. Here, novel multivariate mesoporous MOFs (mMOFs) with regulatable hydrophilic/hydrophobic surfaces were designed by the multivariate competitive strategy and pore modification engineering. 3-Methyl-1H-1,2,4-triazole (3-MTZ) and 5-methyltetrazole (5-MTA) were employed to partially replace 2-MeIm. These were then combined with zinc sulfate heptahydrate (soft templates) to yield mMOFs in a methanol solution. As a proof-of-concept application, we used mMOFs as carriers for enzyme immobilization and investigated the properties of the immobilized enzymes. Benefiting from their mesoporous structure, hydrophilic surface, and improved microenvironment, multivariate mMOFs exhibit a strong ability to stabilize enzyme conformation and increase enzyme activity compared with traditional ZIF-8. Our study offers an avenue for the controllable preparation of well-designed MOF structures, which will further broaden the application opportunities of MOF materials for enzyme immobilization.

Tannic acid mediated hollow biocatalytic zeolitic imidazolate frameworks as a “three-in-one” label for enhancing the sensitivity of fluorescence immunosensor

Benefiting from the special structure feature, zeolitic imidazolate frameworks (ZIFs) have emerged as a carrier for enzyme immobilization. However, the sensitivity of the immunosensor with ZIFs needs further enhancement. Herein, a “three-in-one” label mediated by tannic acid (TA) was designed, demonstrating high enzyme activity, cascade signal amplification capability, and high coupling effectivity with antibodies. The acid property of TA was used as an etchant to etch ZIFs in situ for the generation of hollow ZIFs, which retained the conformational freedom of glucose oxidase for preserving their excellent catalytic activity. The TA on the surface of ZIFs coordinated Fe3+ ions through its phenol groups, forming metal polyphenol network films, which possessed peroxidase-like activity for cascade signal amplification. Meanwhile, the large amount of catechol groups of TA improved the coupling efficiency of antibodies. Based on the proposed “three-in-one” label, a novel fluorescence immunosensor was developed for the sensitive detection of Escherichia coli O157:H7. The limit of detection of the fluorescence immunosensor was 1.35 × 103 CFU mL−1 and 12.51-fold lower than that of traditional immunoassay. This proposed strategy provides a convenient method for preparing hollow ZIFs, facilitating the applications of ZIFs in the fields of biosensors.

Green biocatalyst for decolorization of azo dyes from industrial wastewater: Coriolopsis trogii 2SMKN laccase immobilized on recycled brewer's spent grain.

This study presents an innovative approach for the reuse and recycling of waste material, brewer's spent grain (BSG) for creating a novel green biocatalyst. The same BSG was utilized in several consecutive steps: initially, it served as a substrate for the cultivation and production of laccase by a novel isolated fungal strain, Coriolopsis trogii 2SMKN, then, it was reused as a carrier for laccase immobilization, aiding in the process of azo dye decolorization and finally, reused as recycled BSG for the second successful laccase immobilization for six guaiacol oxidation, contributing to a zero-waste strategy. The novel fungal strain produced laccase with a maximum activity of 171.4 U/g after 6 days of solid-state fermentation using BSG as a substrate. The obtained laccase exhibited excellent performance in the decolorization of azo dyes, both as a free and immobilized, at high temperatures, without addition of harmful mediators, achieving maximum decolorization efficiencies of 99.0%, 71.2%, and 61.0% for Orange G (OG), Congo Red, and Eriochrome Black T (EBT), respectively. The immobilized laccase on BSG was successfully reused across five cycles of azo dye decolorization process. Notably, new green biocatalyst outperformed commercial laccase from Aspergillus spp. in the decolorization of OG and EBT. GC-MS and LC-MS revealed azo-dye degradation products and decomposition pathway. This analysis was complemented by antimicrobial and phytotoxicity tests, which confirmed the non-toxic nature of the degradation products, indicating the potential for safe environmental disposal.

Switchable CO2-Responsive Janus Nanoparticle for Lipase Catalysis in Pickering Emulsion.

The use of convertible immobilized enzyme carriers is crucial for biphasic catalytic reactions conducted in Pickering emulsions. However, the intense mechanical forces during the conversion process lead to enzyme leakage, affecting the stability of the immobilized enzymes. In this study, a CO2-responsive switchable Janus (CrSJ) nanoparticle (NP) was developed using silica NP, with one side featuring aldehyde groups and the other side adsorbing N,N-dimethyldodecylamine. A switchable Pickering emulsion catalytic system for biphasic interface reactions was prepared by covalently immobilizing lipase onto the CrSJ NPs. The CO2-responsive nature of the CrSJ NPs allowed for rapid conversion of the Pickering emulsion, and covalent immobilization substantially reduced lipase leakage while enhancing the stability of the immobilization during the conversion process. Impressively, after repeated transformations, the Pickering emulsion still maintains its original structure. Following 10 consecutive cycles of esterification and hydrolysis reactions, the immobilized enzyme's activity remains at 77.7 and 79.5% of its initial activity, respectively. The Km of the CrSJ catalytic system showed no significant change compared to the free enzyme, while its Vmax values were 1.2 and 1.6 times that of the free enzyme in esterification and hydrolysis reactions, respectively.

Tuning Mechanical Characteristics and Permeability of Alginate Hydrogel by Polyvinyl Alcohol and Deep Eutectic Solvent Addition.

Alginate-based hydrogels are widely utilized for various applications, including enzyme immobilization and the development of drug delivery systems, owing to their advantageous characteristics, such as low toxicity, high availability and cost-effectiveness. However, the broad applicability of alginate hydrogels is hindered by their limited mechanical and chemical stability, as well as their poor permeability to hydrophobic molecules. In this study, we addressed the mechanical properties and chemical resistance of alginate hydrogels in a high-pKa environment by the copolymerization of alginate with polyvinyl alcohol (PVA). The addition of PVA resulted in a threefold improvement in the shear modulus of the copolymeric hydrogel, as well as enhanced chemical resistance to (S)-α-methylbenzylamine, a model molecule with a high pKa value. Furthermore, we addressed the permeability challenge by introducing a betaine-propylene glycol deep eutectic solvent (DES) into the PVA-alginate copolymer. This led to an increased permeability for ethyl 3-oxobutanoate, a model molecule used for bioreduction to chiral alcohols. Moreover, the addition of the DES resulted in a notable improvement of the shear modulus of the resulting hydrogel. This dual effect highlights the role of the DES in achieving the desired improvement of the hydrogel as an immobilization carrier.

Lipase in-situ immobilized in covalent organic framework: Enzymatic properties and application in the preparation of 1, 3-dioleoyl-2-palmitoylglycerol

In this study, the critical importance of designing an appropriate immobilized carrier and method for free lipase to ensure exceptional biological catalytic activity and stability was emphasized. Covalent organic frameworks (COF-1) were synthesized as a novel porous carrier with an azine structure (-CN-NC-) through the condensation of hydrazine hydrate and benzene-1,3,5-tricarbaldehyde at room temperature. Simultaneously, Rhizomucor miehei lipase (RML) was immobilized within the COF-1 carrier using an in-situ aqueous phase method. Characterization of the carrier and RML@COF-1 and evaluation of the lipase properties of RML and RML@COF-1 through p-Nitrophenyl palmitate hydrolysis were conducted. Additionally, application in the synthesis of 1, 3-dioleoyl-2-palmitoylglycerol (OPO) was explored. The results showed that RML@COF-1 exhibited a high enzymatic loading of 285.4 mg/g. Under 60℃ conditions, the activity of RML@COF-1 was 2.31 times higher than that of free RML, and RML@COF-1 retained 77.25% of its original activity after 10 cycles of repeated use, indicating its excellent thermal stability and repeatability. Under the optimal conditions (10%, 1:8 PPP/OA, 45℃, 5 h), the yield of OPO reached 47.35%, showcasing the promising application prospects of the novel immobilized enzyme synthesized via in-situ aqueous phase synthesis for OPO preparation.

Analysis of the feasibility of using biopolymers of different viscosities as immobilization carriers for laccase in synthetic dye removal

The main aim of the study was to assess the feasibility of using biopolymers of different viscosities (high, medium and low viscosity) as immobilization carriers for laccase in synthetic dye removal. The following dye solutions were decolorized: indigo carmine (IC, anionic dye), methylene blue (MB, cationic dye), and their mixture in a molar mass ratio MB/IC=0.69, using biopolymers of different viscosities as laccase immobilization carriers. Toxicity tests were also carried out to assess the toxicity of the post-decolorization samples. Decolorization tests showed that the main decolorization mechanism depends on the dye class. The removal of IC (max. total removal efficiency 72.15%) was mainly by biocatalysis. The mechanism of the MB decolorization process was mainly by sorption on alginate beads, and the efficiency of enzymatic removal was low. However, the highest efficiency of MB decolorization (45.80%) was obtained for beads prepared using the high viscosity alginate when decolorization occurred by both sorption and biocatalysis. The results of mixture decolorization tests differ from the results obtained for single dyes.The results showed differences in the efficiency of the dye sorption process depending on the alginate used for immobilization. Moreover, the varying mechanisms of dye removal from the dye mixture were confirmed by toxicity tests. The occurrence of both biocatalysis and sorption promotes reduced toxicity

Preparation of microbial agent immobilized composites for Cr(VI) removal from wastewater

ABSTRACT Because of its extreme toxicity and health risks, hexavalent chromium [Cr(VI)] has been identified as a major environmental contaminant. Bioreduction is considered as one of effective techniques for cleaning up Cr(VI)-contaminated sites, but the remediation efficiency needs to be enhanced. Here, a novel immobilized microbial agent was produced by immobilizing Bacillus cereus ZY-2009 with sodium alginate (SA) using polyvinyl alcohol (PVA) and activated carbon (AC). To evaluate the decrease of Cr(VI) by immobilized bacterial agents, batch tests were conducted with varying immobilization conditions, immobilization carriers, and dosages of medication. The removal of Cr(VI) by the agent prepared by the composite immobilization method was better than that by the adsorption and encapsulation methods. The optimal preparation conditions were the fraction of magnetic PVA was 5.00%, the fraction of SA was 4.00%, the fraction of CaCl2 was 4.00%, and the calcification time was 12 h. The experimental results indicated that PVA/SA/AC agents accelerated the reduction rate of Cr(VI). The removal rate of Cr(VI) by immobilized cells (90.5%) under ideal conditions was substantially higher than that of free cells (11.0%). This novel agent had a large specific surface area and a rich pore structure, accounting for its high reduction rate. The results suggest that the PVA/SA/AC immobilized Bacillus cereus ZY-2009 agent has great potential to remove Cr(VI) from wastewater treatment systems.

The remediation of polycyclic aromatic hydrocarbon contaminated soil by immobilized microorganisms using distiller's grains.

Polycyclic aromatic hydrocarbons are a persistent organic pollutant, and their biodegradation in the soil is often limited due to the limited degradation ability of indigenous bacteria and the low activity of exogenous PAH degrading bacteria. Immobilized microbial technology can protect microorganisms from the impact of harsh environments, and distiller's grains have the potential as carriers for microbial immobilization. This study aims to use distiller's grains as a microbial carrier, investigate the feasibility of immobilized microorganisms using distiller's grains for remediation of PAH contaminated soil; explore the relationship between soil nutrient content, consumption, and PAH degradation rate; and reveal the mechanism of bioremediation from the perspective of soil enzyme activity and microbial community composition. The results showed that after 72 days of remediation, the removal rates of phenanthrene and pyrene in the treatment of immobilized microorganisms in distiller grains reached 91.78% and 58.59%, respectively. Distiller grains can serve as a carrier for microorganisms, providing them with shelter and nutrients to enhance their chance of survival. Additionally, they can regulate the composition of soil particles and improve aeration, thereby increasing the efficiency of PAH degradation in soil.

Coupling chemical modification and immobilization of cellulase improves thermal stability and low-melting mixture solvent tolerance for in situ saccharification of bagasse

In situ saccharification of lignocellulosic biomass in low-melting mixture solvents (LoMMSs) is a promising approach for biorefinery. In this study, chemical modification of cellulase coupled with immobilization was designed to improve thermal stability and LoMMS tolerance of cellulase for in situ saccharification. Cellulase was firstly chemically modified with succinic anhydride and ethylenediamine respectively to form succinylated cellulase (S-Ce) and aminated cellulase (A-Ce), then the modified cellulases were immobilized with amino-functionalized metal-organic frameworks to establish ZIF-8/A immobilized A-Ce and S-Ce. Circular dichroism, fluorescence measurements, X-ray diffraction, fourier transform infrared spectra and scanning electron microscopy were applied to characterize the modified cellulase or immobilization carrier. ZIF-8/A immobilized A-Ce presented desired thermal stability and LoMMS tolerance. Reducing sugar yields via ZIF-8/A immobilized A-Ce during in situ saccharification of bagasse maintained stably till Bet/AA concentration reached 50%, and declined relatively slowly compared with free or other immobilized cellulase when Bet/AA concentration further increased to 90%. In situ saccharification of bagasse via ZIF-8/A immobilized A-Ce in concentrated Bet/AA (50%) can be operated at a wide temperature range (50–60 °C). The corresponding reducing sugar yield with ZIF-8/A immobilized A-Ce was 58% higher than that with free cellulase in 50% Bet/AA at 60 °C.

Mesocellular Silica Foam as Immobilization Carrier for Production of Statin Precursors.

The employment of 2-deoxyribose-5-phosphate aldolase (DERA) stands as a prevalent biocatalytic route for synthesizing statin side chains. The main problem with this pathway is the low stability of the enzyme. In this study, mesocellular silica foam (MCF) with different pore sizes was used as a carrier for the covalent immobilization of DERA. Different functionalizing and activating agents were tested and kinetic modeling was subsequently performed. The use of succinic anhydride as an activating agent resulted in an enzyme hyperactivation of approx. 140%, and the stability almost doubled compared to that of the free enzyme. It was also shown that the pore size of MCF has a decisive influence on the stability of the DERA enzyme.

PVA-TiO2 Nanocomposite Hydrogel as Immobilization Carrier for Gas-to-Liquid Wastewater Treatment

This study investigates the development of polyvinyl alcohol (PVA) gel matrices for biomass immobilization in wastewater treatment. The PVA hydrogels were prepared through a freezing–thawing (F-T) cross-linking process and reinforced with high surface area nanoparticles to improve their mechanical stability and porosity. The PVA/nanocomposite hydrogels were prepared using two different nanoparticle materials: iron oxide (Fe3O2) and titanium oxide (TiO2). The effects of the metal oxide nanoparticle type and content on the pore structure, hydrogel bonding, and mechanical and viscoelastic properties of the cross-linked hydrogel composites were investigated. The most durable PVA/nanoparticles matrix was then tested in the bioreactor for the biological treatment of wastewater. Morphological analysis showed that the reinforcement of PVA gel with Fe2O3 and TiO2 nanoparticles resulted in a compact nanocomposite hydrogel with regular pore distribution. The FTIR analysis highlighted the formation of bonds between nanoparticles and hydrogel, which caused more interaction within the polymeric matrix. Furthermore, the mechanical strength and Young’s modulus of the hydrogel composites were found to depend on the type and content of the nanoparticles. The most remarkable improvement in the mechanical strength of the PVA/nanoparticles composites was obtained by incorporating 0.1 wt% TiO2 and 1.0 wt% Fe2O3 nanoparticles. However, TiO2 showed more influence on the mechanical strength, with more than 900% improvement in Young’s modulus for TiO2-reinforced PVA hydrogel. Furthermore, incorporating TiO2 nanoparticles enhanced hydrogel stability but did not affect the biodegradation of organic pollutants in wastewater. These results suggest that the PVA-TiO2 hydrogel has the potential to be used as an effective carrier for biomass immobilization and wastewater treatment.

Enhancement of enzyme activity by laser-induced energy propulsion of upconverting nanoparticles under near-infrared light: A comprehensive methodology for in vitro and in vivo applications

If the appropriate immobilization method and carrier support are not selected, partial decreases in the activity of enzymes may occur after immobilization. Herein, to overcome this challenge, an excitation mechanism that enables energy transfer was proposed. Modified upconverting nanoparticles (UCNPs) were constructed and the important role of near-infrared (NIR) excitation in enhancing the catalytic activity of the enzyme was demonstrated. For this purpose, UCNPs were first synthesized via the hydrothermal method, functionalized with isocyanate groups, and then, PEG-L-ASNase was immobilized via covalent binding. UCNPs with and without PEG-L-ASNase were extensively characterized by different methods. These supports had immobilization yield and activity efficiency of >96 % and 78 %, respectively. Moreover, immobilized enzymes exhibited improved pH, thermal, and storage stability. In addition, they retained >65 % of their initial activity even after 20 catalytic cycles. Biochemical and histological findings did not indicate a trend of toxicity in rats due to UCNPs. Most importantly, PEG-L-ASNase activity was triggered approximately 5- and 2-fold under in vitro and in vivo conditions, respectively. Overall, it is anticipated that this pioneering work will shed new light on the realistic and promising usage of NIR-excited UCNPs for the immobilization of enzymes in expensive and extensive applications.

Application potential of modified waste-lignin as microbial immobilization carriers for improve soil fertility

The immobilized microorganism technology is used to load the functional microorganism on a suitable carrier to maintain its biological activity, resist the impact of adverse environment. In this study, chemical modification of lignin side chain phenolic hydroxyl groups was carried out to increase the content of amide functional groups and enhance their adsorption and fixation effects on microorganisms due to the numerous active sites in the structure of discarded lignin, and studied its impact on microbial activity and crop growth. The results show that the modified lignin could be effectively combined with functional microorganisms, and the proposed bio-composite material was an effective method for cell protection. In addition, it was shown that functional microorganisms maintained high biological activity by immobilization of modified lignin, which was 34.3–41.0% higher than that of CK. Moreover, the technology could increase the total nitrogen content and available phosphorus content in soil by 70.8% and 64.4%, respectively, compared with CK. Therefore, it could increase eventually wheat growth indices and biomass. The completed research opens new perspectives for the effective immobilization of microorganisms and utilization of wasted lignin of significant economic importance.

Preparation of Ordered Macroporous ZIF-8-Derived Magnetic Carbon Materials and Its Application for Lipase Immobilization

Metal–organic framework materials (MOFs) and their derivatives are considered ideal immobilization carrier materials because of their large specific surface area, high porosity and excellent structural designability. Among them, ZIF-8 has great potential for immobilization of enzymes due to mild synthesis conditions, and good biocompatibility. However, conventional ZIF-8 crystals have poor separation and recovery efficiency due to their small pore size and poor acid stability, greatly limiting their application in enzyme immobilization and further application. Although the carbonization of ZIF-8 by pyrolysis has been shown to be one of the approaches that can enhance its chemical stability, this still does not effectively solve the problem of the difficulty of recycling. Herein, we developed a strategy of pre-carbonization immersion (immersion in aqueous FeSO4 solution before carbonization) to synthesize ordered macroporous ZIF-8-derived carbon materials with stable ferromagnetism (denoted as CZ-x-M-y, where x denotes the carbonization temperature and y denotes the concentration of the impregnated FeSO4 solution) and used them to immobilize lipases for biodiesel production. XRD analysis showed that the magnetic properties in the materials came from Fe3C species. We found that the magnetic carbon materials obtained by carbonization at 600 °C showed the best immobilization effect, where CZ-600-M-0.3 (using 0.3 mol·L−1 FeSO4 aqueous solution to soak ZIF-8 and carbonized at 600 °C) had the highest enzyme loading of 183.04 mg·g−1, which was 49.7% higher than that of the non-magnetic CZ-600. In addition, CZ-600-M-0.5 maintained the highest enzyme activity, which was 81.9% of the initial activity, after five batches of reuse. The stable magnetic support materials reported in this study have promising potential for the industrial application of immobilized lipase.

Pros and Cons in Various Immobilization Techniques and Carriers for Enzymes.

In recent years, enzyme immobilization technology has been developed, and studies on immobilized enzyme materials have become very prominent. With the immobilization technique, enzymes and compatible carrier materials are combined or enzyme crystals/aggregates are used in a carrier-free fashion, by physical, chemical, or biochemical methods. As a kind of biocatalyst, immobilized enzymes can catalyze certain chemical reactions with high selectivity and high efficiency under relatively mild reaction conditions and eliminate pollution to the environment. Considering the current status and applications of immobilized enzyme technology and materials emerging in the last 5years, this mini-review introduces the advantages and disadvantages of various enzyme immobilization techniques with carriers as well as the pros and cons of different materials for immobilization. The future prospects of immobilization technology and carrier materials are outlined, aiming to provide a reference for further research and applications of sustainable technology.

LIPASE FROM Rhizopus oryzae IMMOBILIZED ON CORN COB POWDER: A NEW APPROACH FOR DIETETIC TRIGLYCERIDE SYNTHESIS IN A FIXED BED REACTOR

This work explores the possibility of using corn cob powder as a carrier for immobilization by physical adsorption of lipase from Rhizopus oryzae used in acidolysis reaction of grape seed oil with capric acid (C10) to synthesize dietary triglycerides (TAGs) formed from medium-long-medium chain (MLM) fatty acids in a fixed bed reactor. First, enzymatic loading was investigated to improve hydrolytic activity and the immobilized biocatalyst was characterized in terms of biochemical parameters and thermal stability. An enzyme load of 30 mg mL-1 was quite adequate, as about 90% of the immobilization yield was achieved. The central composite rotatable design (CCRD) experimental design revealed that, for the immobilized enzyme, the optimal hydrolytic activity was obtained at pH 7.75 and 41 °C. Stirred tank reactions were optimized in CCRD statistical analysis revealing the most desirable conditions for molar ratio of 1:9.36 (oil:acid), 45 °C and 15% biocatalyst, obtaining a maximum ID value of 86.93 ± 2.25%. In a fixed bed reactor, the synthesis showed an average ID of 51.13 ± 0.77, demonstrating the potential of this configuration. It is concluded that corn cob powder as an immobilization carrier is a promising alternative for the enzymatic synthesis of MLM triacylglycerols.