Enzyme Carrier Research Articles

Biochar as an Enzyme Immobilization Support and Its Application for Dye Degradation

Wastewaters generated by the textile industry often contain significant amounts of harmful (carcinogenic and mutagenic) cationic dyes, whose efficient removal is of crucial importance. This study investigates the laccase immobilization on biochar obtained from sour cherry stones (SCS-B), as a cost effective adsorbent, and evaluates its application for brilliant green (BG) degradation. The successful immobilization of laccase on biochar was achieved via adsorption and confirmed using scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX) and Fourier transform infrared spectroscopy (FTIR). An immobilization efficiency of 66% was achieved using 0.274 U/mL of laccase at pH 5 and a temperature of 40 °C. The adsorption kinetics of laccase followed a pseudo-second-order model, indicating that chemical adsorption plays a significant role in the immobilization process. The BG degradation by immobilized system was further optimized by evaluating effects of pH, temperature, dye concentration, and contact time. More than 92% of BG (50 mg/L) was removed within 4 h at pH 5 and temperature of 30 °C. These findings suggest that SCS-B can effectively be used as an enzyme carrier and be further utilized for the removal of emerging pollutants, positioning it as a sustainable solution for wastewater treatment.

Dispersion of halloysite nanotube/lipase nanohybrids as nanofillers into polyvinyl alcohol-sodium alginate cryogel: Characterization and bio-catalytic activity analysis

The purpose of this study is to formulate and characterize the cryogels containing halloysite nanotube (HNT)/lipase nanohybrid (NH-cryogel) in comparison to pure cryogels as well as cryogels containing lipase (lipase-cryogel). The cryogels were synthesized using polyvinyl alcohol (PVA) and sodium alginate (SA). The products are tested to explore the influence of the HNT/lipase nanohybride (NH) as nanofillers on the cryogel properties using methods such as swelling degree, water uptake measurement, TGA, XRD, FESEM and FTIR. Additionally, the effects of cryogels on the stability and biocatalytic activities of lipase and NH, were studied and compared to the free lipase to evaluate their potential applications as enzyme carriers. The addition of nanofillers into the cryogel improved is thermal stability. The results implied that NH-cryogel had better enzyme activity than lipase-cryogel and free lipase at different temperatures and pH values. The NH-cryogel residual activity was 85.5 % after ten cycles of reuse while lipase-cryogel showed lower residual activity (60.3 %). Furthermore, the NH-cryogel retained 81.1 % of its residual activity while this was 51.0 % for lipase-cryogel after thirty days of storage. Therefore, the presented results in this study provide a pathway to show that produced nano-composite cryogels could be useful substances for food and pharmaceutical industries applications.

Hydrophobic Ionic Liquid Gel Microspheres as Bi‐Component Carriers with a Liquid Phase to Immobilize Enzymes for Enhanced Performance

AbstractThis study focuses on incorporating liquid molecules, different from the bulk solution, into the immobilized enzyme carrier to regulate the distribution effect and diffusion‐limiting impact of the carrier's microenvironment for substrates, which generally is difficult to achieve due to the instability of the materials with liquid inclusions. A freestanding liquid‐holding particle carrier, with bi‐component hydrophobic ionic liquid gel microspheres with poly (glycidyl methacrylate) as the network and 1‐butyl‐3‐methylimidazolium hexafluorophosphate as the dispersing medium, is proposed, which is stable in air aqueous solution, and can extract proteins and organic small molecules into its interior due to the mobility of its dispersing medium. Horseradish peroxidase is covalently immobilized into the microspheres, forming a liquid compartment enzyme microreactor. The microreactor exhibits superior stability, enzymatic activity, and catalytic performance for Basic Orange II degradation compared to free enzyme and liquid‐free immobilized enzymes. This is attributed to the biocompatibility of the ionic liquid, its role in substrate enrichment in its interior, and its rapid mass transfer capability. This contribution shows the effectiveness of regulating the carrier's microenvironment with liquid molecules, offering fresh perspectives and strategies for enzyme technology.

Natural shellac-based microcapsules as lipase carriers for recyclable efficient Pickering interfacial biocatalysis

Enzyme is an important class of catalyst. However, the efficiency of enzyme-catalyzed reactions is constrained by the limited contact between the enzyme and its substrate. In this study, to overcome this challenge, lipase-loaded microcapsules were prepared from natural shellac and nanoparticles using the emulsion template method. These microcapsules can perform dual roles as stabilizers and enzyme carriers to construct a water-in-oil Pickering interfacial biocatalytic system. The results showed that the hydrolytic conversion of the microcapsules could reach 90% within 20 min, which was significantly higher than that of the traditional biphasic system. The catalytic activity was influenced by the oil-to-water volume ratio and the microcapsule content. The microcapsules remained highly catalytic efficiency even after storage for three months or seven cycles of reuse. These microcapsules were prepared without the use of any cross-linkers or harsh solvents. This green and efficient catalytic system has great application prospects in the food industry.

Optimizing Continuous‐Flow Biocatalysis with 3D‐Printing and Inline IR Monitoring

AbstractEnzymatic biocatalysis typically generates less waste, uses less water, and minimizes energy consumption compared to traditional chemical methods. Efficient, cell‐free biosynthesis relies on the reuse of its valuable biocatalysts. Immobilization of enzymes on solid supports, such as enzyme carrier resins (ECRs), offers a reliable and widely deployed approach to maximize enzyme turnover in cell‐free biosynthesis. We focus on two major bottlenecks associated with optimizing cell‐free biocatalysis. First, we apply our lab's 3D‐printed labware to screen ECRs in 96‐well mini‐reactors to optimize enzyme immobilization conditions. Second, we introduce inline infrared spectroscopy to monitor bioreactor output and maximize enzyme productivity. Urease provides a model system for examining immobilization conditions and continuous assessment of biocatalyst performance. As required for the high substrate concentrations to improve process efficiency and minimize waste, urease was studied in unusually high concentrations of its substrate – molar concentrations of urea. The optimized reactor processed 3.24 L of 4.00 M urea at an average volumetric productivity of 13 g ⋅ L−1 ⋅ h−1 over 18 h and achieved an estimated productivity number of >17.4 kg urea processed per g of immobilized urease Type‐IX. This workflow can be generalized to most biocatalytic processes and could accelerate adoption of cell‐free biosynthesis for greater chemical sustainability.

Colorimetric Immunoassays with Boronic Acid-Decorated, Peroxidase-like Metal-Organic Frameworks as the Carriers of Antibodies and Enzymes.

The sensitivity of immunoassays is generally limited by the low signal reporter/recognition element ratio. Nanomaterials serving as the carriers can enhance the loading number of signal reporters, thus improving the detection sensitivity. However, the general immobilization strategies, including direct physical adsorption and covalent coupling, may cause the random orientation and conformational change in proteins, partially or completely suppressing the enzymatic activity and the molecular recognition ability. In this work, we proposed a strategy to load recognition elements of antibodies and enzyme labels using boronic acid-modified metal-organic frameworks (MOFs) as the nanocarriers for signal amplification. The conjugation strategy was proposed based on the boronate ester interactions between the carbohydrate moieties in antibodies and enzymes and the boronic acid moieties on MOFs. Both enzymes and MOFs could catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by H2O2, therefore achieving dual signal amplification. To indicate the feasibility and sensitivity of the strategy, colorimetric immunoassays of prostate specific antigen (PSA) were performed with boronic acid-modified Cu-MOFs as peroxidase mimics to catalyze TMB oxidation and nanocarriers to load antibody and enzyme (horseradish peroxidase, HRP). According to the change in the absorbance intensity of the oxidized TMB (oxTMB), PSA at the concentration range of 1~250 pg/mL could be readily determined. In addition, this work presented a site-specific and oriented conjugation strategy for the modification of nanolabels with recognition elements and signal reporters, which should be valuable for the design of novel biosensors with high sensitivity and selectivity.

Nanosponges: A Targeted Drug Delivery System

The latest advances in nanotechnology have resulted in the development of a targeted drug delivery system. However, in order to efficiently target a molecule to a specific place with a drug delivery system, a specialized drug delivery system is required. The discovery of nanosponge has been a big step toward overcoming challenges such as drug toxicity, low bioavailability, and predictable drug release because they can accept both hydrophilic and hydrophobic drugs. Nanosponges, a newly developed colloidal system, have the potential to address concerns such as medicine toxicity, lower bioavailability, and drug release over a large region because they can be adjusted to operate with both hydrophilic and hydrophobic pharmaceuticals. Nanosponges are small structures with a three-dimensional network and porous hollow. Nanosponges are small structures with a three-dimensional network and porous hollow. They can be easily created by crosslinking cyclodextrins with various chemicals. Because of Cyclodextrin's excellent biocompatibility, stability, and safety, a number of Cyclodextrin-based drug delivery systems have been rapidly developed. The nanosponge drug delivery system has a wide range of applications, including cancer, autoimmune illnesses, theranostic uses, increased bioavailability, and stability. This review delves into the benefits and downsides, preparation procedures, factors influencing their preparation, characterisation techniques, applications, and the most recent advancements in nanosponges. Nanosponges can also act as an efficient carrier of enzymes, proteins, vaccines, and antibodies. The current review focuses on the method of preparation, characterisation, and possible application in drug delivery systems.

Preparation and characterization of AChE immobilized magnetic bio-nanocomposites (Fe3O4@Cht/Au) for pesticide detection

Free enzymes cause difficulties in many applications due to their insufficient stability, loss of activity in a short time, and most importantly, although they are costly, they are used only once in reactions, lose their effect and cannot be recovered from the environment. Magnetic nanoparticles coated with biocompatible polymeric material are potential candidates for promising enzyme carriers due to their multifunctional pore surfaces, easy removal from the environment provided by the magnetization, ability to main stability under various harsh conditions. This study prepared a biosensor candidate based on the inhibiting acetylcholinesterase enzyme by organophosphate pesticides from chitosan-coated magnetic nanoparticles doped with gold. Transmission electron microscopy, scanning electron microscopy, X-ray diffraction diffractometry, and Fourier transform infrared spectroscopy analysis confirmed the structure of synthesized nanocomposites. Magnetic characteristics of the nanocomposites were assessed using VSM. Bio-nanocomposite (Fe3O4@Cht/Au/AChE) was used to determine environmental pollutants qualitatively. Remediation of organophosphate-containing wastewater is an essential issue for environmental sustainability. In this work, Dichlorvos and Chlorpyrifos were selected as organic pollutants to assess the enzymatic activity of immobilized Fe3O4@Cht/Au/AChE. Optimum conditions for AChE enzyme were immobilized nanostructures (Fe3O4@Cht/Au/AChE) were determined. The optimum pH for the immobilized enzyme was found to be 8, and the optimum temperature was found to be 60 °C. Retained immobilized enzyme activity is found to be around 50% for the 20th reuse. In the presence of 150 µL pesticide, retained immobilized enzyme activity is found to be around 25%. Method validation was performed for pesticides. When using immobilized AChE, the LOD (limit of detection)-LOQ (limit of quantitation) values for Dichlorovos and Chlorpyrifos was obtained in the range of 0.0087–0.029 nM and 0.0014–0.0046 nM, respectively. The relative standard deviation (RSD%) values, which are indicators of precision, were found to be below 2%.

Heteropolysaccharide-Based Coacervate Microdroplets as Enzyme Carriers for Detection of Cholesterol

Heteropolysaccharide-Based Coacervate Microdroplets as Enzyme Carriers for Detection of Cholesterol

Magnetic biocatalytic nanoreactors based on graphene oxide with graded reduction degrees for the enzymatic synthesis of phytosterol esters

As we know, the carriers of enzymatic nanoreactors with different amphipathy have significant influence on the interfacial enzyme activities and its performance. In this study, the magnetic graphene oxide (GO) enzyme carriers were constructed by assembling graphene oxide with different reduction degrees (rGO(x)) and polyethyleneimine (PEI) modified nano-flowery Fe3O4 into composite materials through electrostatic interactions, and x here represents the reduction reaction hours of GO. The SEM, TEM, contact angles and BET tests qualified the ideal enzyme carrier, PEI-Fe3O4-rGO(12), by adsorption-crosslinking method, the Candida rugosa lipase (CRL) was immobilized to obtain the lipase nanoreactor, PEI-Fe3O4-rGO(12)@CRL. The nano-sheet and nano-flower structures provided a larger specific surface area and suitable pore size, and the CRL had higher protein load on the carrier material (163.11 mg/g). In addition, using hydrolyzed p-nitrophenol palmitate as the model, the PEI-Fe3O4-rGO(12)@CRL can still show 76.5 % residual activity after 8 repeated uses. At the same time, the immobilized lipase showed good catalytic performance in the biosynthesis of phytosterol esters. The optimal esterification conditions were determined by single factor experiment (isooctane, sterol concentration 150 μmol/mL, alkyd molar ratio 1:2, enzyme dosage 120 mg, 26 h, 55 °C, the esterification rate was 76.24 %). Therefore, the optimal biocatalyst PEI-Fe3O4-rGO(12)@CRL may has good performance in the biosynthesis of other novel foods.

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.

Evaluation of free and immobilized cellulase on chitosan-modified magnetic nanoparticles for saccharification of sorghum residue.

Enzymatic hydrolysis plays a pivotal role in transforming lignocellulosic biomass. Addressing alternate techniques to optimize the utilization of cellulolytic enzymes is one strategy to improve its efficiency and lower process costs. Cellulases are highly specific and environmentally benign biocatalysts that break down intricate polysaccharides intosimple formsofsugars. In contrast to the most difficult and time-consuming enzyme immobilization processes, in this research, we studied simple, mild, and successful techniques for immobilization of pure cellulase on magnetic nanocomposites using glutaraldehyde as a linker and used in the application of sorghum residue biomass. Fe3O4 nanoparticles were coated with chitosan from the co-precipitation method, which served as an enzyme carrier. The nanoparticles were observed under XRD, Zeta Potential, FESEM, VSM, and FTIR. The size morphology results presented that the Cs@Fe3O4 have 42.2nm, while bare nanoparticles (Fe3O4) have 31.2nm in size. The pure cellulase reaches to 98.07% of loading efficiency and 71.67% of recovery activity at optimal conditions. Moreover, immobilized enzyme's pH stability, thermostability, and temperature tolerance were investigated at suitable conditions. The kinetic parameters of free and immobilized enzyme were estimated as Vmax; 29 ± 1.51 and 27.03 ± 2.02µmolmin-1mg-1, Km; 4.7 ± 0.49mM and 2.569 ± 0.522mM and Kcat; 0.13s-1, and 0.89s-1. Sorghum residue was subjected to 2% NaOH pre-treatment at 50℃. Pre-treated biomass contains cellulose of 64.8%, used as a raw material to evaluate the efficiency of reducing sugar during hydrolysis and saccharification of free and immobilized cellulase, which found maximum concentration of glucose 5.42g/L and 5.12g/L on 72h. Thus, our study verifies the use of immobilized pure cellulase to successfully hydrolyze raw material, which is a significant advancement in lignocellulosic biorefineries and the reusability of enzymes.

Functionalized magnetic nanoparticles for Cellic Ctec2 cellulase immobilization: Allowing reusability of enzyme in the conversion of cellulosic biomass

The method of enzyme immobilization can ameliorate the overall stability and restoration of enzymes, hence facilitating their broader application in several sectors. This investigation utilized cellulase as a hydrolytic enzyme. In order to enhance the stability and performance of the cellulase enzyme, the research employed immobilization technology to secure the Cellic Ctec2 cellulase to the synthesized Cs@Fe3O4 nanocomposites. Fe3O4 nanoparticles (NPs) were coated with chitosan obtained from co-precipitation method that served as enzyme carrier. The NPs (Cs@ Fe3O4) were observed under XRD; VSM (vibrating-sample magnetometer) shows saturation magnetizations (Ms), UV–vis, field emission scanning electron microscopy (FESEM), and Fourier transform infrared spectroscopy (FT-IR). Response surface approach was applied to optimize the conditions for immobilization of cellulase. The optimum immobilization of cellulase reaches to 99.1% of loading efficiency and 69.7% of recovery activity with 2.5% of glutaraldehyde concentration. Furthermore, under ideal circumstances the immobilized enzyme’s thermostability, pH stability, temperature tolerance and reusability, were studied with respect to free cellulase. Higher relative activity of cellulase enzyme was observed at pH 5 with 50 °C temperature than free enzyme. One percent CMC hydrolysis is considered for reusability of free and immobilized enzyme and releases 222 mg glucose/g substrate at 24 h, showing great quiescence in cellulosic biomass conversion. Immobilized cellulase demonstrated high reusability by retaining almost 61.2% up to the 5th cycles and 51.2% of activity-maintained 10th cycle of hydrolysis. Reusability of cellulase enzyme can attain a gradual decrease in relative activity as number of repeats of the cycle increases to 10 during hydrolysis and increases in glucose yield after hydrolysis.

Elucidation of enhanced cellulase immobilization onto synthetic magnetic nickel nanomaterials for lignocellulosic biomass hydrolysis

The immobilization of cellulase enzymes for the conversion of lignocellulosic biomass into sustainable biochemical products is essential for the stability and recovery of the enzymes. In this study, nickel nanoparticles (NiNPs) were synthesized and coated with 3-aminopropyl triethoxysilane (APTES) to serve as cellulase enzyme carriers. Cellulase enzyme immobilization on the prepared NiNPs was achieved using glutaraldehyde as a cross-linker. The physicochemical properties of the carrier were determined before and after enzyme immobilization using X-ray Powder Diffraction (XRD), Transmission Electron Microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), Vibrating-Sample Magnetometer (VSM), and Field Emission Scanning Electron Microscopy (FESEM). The impacts of different experimental factors on the performance of the cellulase enzymes immobilized on the synthesized NiNPs and commercial nickel nanoparticles were compared. The results showed comparable optimal cellulase enzyme immobilization conditions on both substrates in terms of the immobilization time, pH, and cellulase enzyme concentration. However, the recommended temperatures for cellulase enzyme immobilization on the synthesized and commercial NiNPs were 50 °C and 40 °C, respectively. Under these optimum conditions, the immobilized cellulase enzyme on the synthesized NiNPs had an activity of about 99.1% (in comparison to the activity of free cellulase enzyme), while the activity of the enzyme upon immobilization on commercial NiNPs was about 93%. The particle size of the NiNPs was found to be crucial for enzyme immobilization efficiency and its magnetic strength. Therefore, cellulase enzyme immobilization on tunable NiNPs could be a sustainable and eco-friendly approach towards high recovery of cellulose from lignocellulosic materials.

Nanomaterials for co‐immobilization of multiple enzymes

AbstractIn order to co‐immobilize multiple enzymes, a wide range of nanomaterials has been designed to achieve synergistic enzyme activity and enhance catalytic efficiency. Nanomaterials, as carriers for enzyme co‐immobilization, possess various advantages such as tunable morphology and size, high specific surface area, and abundant chemically active sites. They can significantly enhance enzyme stability, activity, and catalytic efficiency. We overview the commonly used methods and strategies of enzyme co‐immobilization. This review further summarizes the latest research advances in nanomaterials for enzyme co‐immobilization applications over the past 5 years. Meanwhile, the advantages and challenges of these nanomaterials used for enzyme co‐immobilization as well as some potential future directions are also discussed.

Effects of Enzyme Treatment and Carrier Agents on Chemical and Physical Properties of Almond Protein-Based Product

There has been an emerging trend towards the research of plant-based proteins over the past few years; however, there has been limited conductive observation of plant-based proteins from almond by-products, as well as the potential of using Flavourzyme and different carrier agents to obtain an instant protein-based powder. The purpose of this study was to investigate the effects of Flavourzyme at various concentrations (0, 0.5, 1, 1.5, 2, and 2.5%) and incubation times (30, 60, 90, 120, and 180 min), as well as carrier agents at different ratios with a 20% total concentration on the physicochemical properties of almond protein-based products using freeze-drying (FD). The results showed a higher protein content (p-value ≤ 0.05) using 1.5% Flavourzyme for 120 min compared to the other levels. There was no significant difference (p-value > 0.05) in the protein retention rate after freeze-drying of the samples. However, the addition of Maltodextrin (MA), Gum Arabic (GA), and Inulin (IN) encapsulants improved the physical and functional characteristics of freeze-dried almond protein-based powder (FDAP). However, the solubility of the powder is moderate. Water-holding-capacity (WHC) and Oil- Holding Capacity (OHC) are inversely proportional, where GA-coated powder is the most hydrophobic (17.72 ± 0.87 mL oil/g) and IN-encapsulated powder is the most hydrophilic (15.00 ± 0.87 mL water/g). In conclusion, IN could be a potential encapsulant for almond protein-based powder, because powder produced from IN or MA: IN is acceptable in terms of physical parameters, and IN can also enhance fiber content in the final product.

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.

Covalently crosslinked coacervates: immobilization and stabilization of proteins with enhanced enzymatic activity.

Coacervates represent models for membrane-free protocells and thus provide a simple route to synthetic cellular-like systems that provide selective encapsulation of solutes. Here, we demonstrate a simple and versatile post-coacervation crosslink method using the thiol-ene click reaction in aqueous media to prepare covalently crosslinked coacervates. The crosslinking of the coacervate enables stability at extreme pH where the uncrosslinked coacervate fully disassembles. The crosslinking also enhances the hydrophobicity within the coacervate environment to increase the encapsulation efficiency of bovine serum albumin (BSA), as compared to the uncrosslinked coacervate. Additionally, the crosslinked coacervate increases the stabilization of BSA at low pH. These crosslinked coacervates can act as carriers for enzymes. The enzymatic activity of alkaline phosphatase (ALP) is enhanced within the crosslinked coacervate compared to the ALP in aqueous solution. The post-coacervation crosslink approach allows the utilization of coacervates for encapsulation of biologicals under conditions where the coacervate would generally disassemble. We demonstrate that these crosslinked coacervates enable the protection of encapsulated protein against denaturation at extreme pH and enhance the enzymatic activity with encapsulation. This click approach to stabilization of coacervates should be broadly applicable to other systems for a variety of biologics and environmentally sensitive molecules.

Synthesis of microfibrillated cellulose from solid residue of seaweed processing industry and its applications in alginate-based hydrogels for papain enzyme carriers

Synthesis of microfibrillated cellulose from solid residue of seaweed processing industry and its applications in alginate-based hydrogels for papain enzyme carriers

A lipase-conjugated carbon nanotube fiber-optic SPR sensor for sensitive and specific detection of tributyrin.

As a low-density lipoprotein, tributyrin plays an essential role in food safety and human health. In this study, a novel lipase-conjugated carbon nanotube (CNT) surface plasmon resonance (SPR) fiber-optic sensor is used to specifically detect tributyrin for the first time. In this work, CNTs can be used as an amplifying material to significantly increase the sensitivity of SPR sensors due to their high refractive index and large surface area. CNTs can also be used as an enzyme carrier to provide abundant carboxyl groups for the specific binding of lipases. Covering the surface of the sensor with CNTs can not only enhance the performance of the sensor, but also provide sufficient detection sites for subsequent biomass detection, reduce the functionalization steps, and simplify the sensor preparation process. The experimental results demonstrate that the refractive index sensitivity of the traditional multimode fiber (MMF)-single mode fiber (SMF)-MMF transmissive optical fiber sensor is 1705 nm RIU-1. After covering the sensor with CNTs, the sensitivity is 2077 nm RIU-1, and the sensitivity has been improved very well. In addition, there are abundant functional groups on CNTs, which can provide abundant binding sites. Conjugating lipase on carbon nanotubes helps to achieve linear detection in the range of 0.5 mM to 4 mM tributyrin, with a sensitivity of 4.45 nm mM-1 and a detection limit of 0.34 mM, which is below the 2.26 mM detection standard and meets food safety monitoring requirements. Compared with other sensors, the optical fiber biosensor proposed in this study expands the concentration detection range of tributyrin. Furthermore, the sensor also has good stability, anti-interference performance and specificity. Therefore, the sensor proposed in this paper has good application prospects in the fields of food safety and biomedicine.