Burkholderia Cepacia Lipase Research Articles

Design for preparation of more active cross-linked enzyme aggregates of Burkholderia cepacia lipase using palm fiber residue.

A new design of cross-linked enzyme aggregates (CLEAs) of Burkholderia cepacia lipase (BCL) based mainly on the use of lignocellulosic residue of palm fiber as an additive was proposed. Different parameters for the preparation of active CLEAs in the hydrolysis of olive oil, such as precipitation agents, crosslinking agent concentration, additives, and coating agents were investigated. The highest activity yield (121.1 ± 0.1%) and volumetric activity (1578.1 ± 2.5 U/mL) were achieved for CLEAs prepared using the combination of a coating step with Triton® X-100 and polyethyleneimine plus the use of palm fiber as an additive. The variations of the secondary structures of BCL-CLEAs were analyzed by second-derivative infrared spectra, mainly indicating a reduction of the α-helix structure, which was responsible for the lipase activation in the supramolecular structure of the CLEAs. Thus, these results provided evidence of an innovative design of BCL-CLEAs as a sustainable and biocompatible opportunity for biotechnology applications.

Enantioselective synthesis of a chiral intermediate of himbacine analogs by Burkholderia cepacia lipase A.

The enantiomers of (4R/S)-4-hydroxy-N, N-diphenyl-2-pentynamide are key chiral synthons for the synthesis of thrombin receptor antagonists such as vorapaxar. In this paper, we report the enzymatic preparation of enantiomerically enriched (4R)-4-hydroxy-N, N-diphenyl-2-pentynamide using lipase A from Burkholderia cepacia ATCC 25416 as the catalyst. First, the lipase gene (lipA) and its chaperone gene (lipB) was cloned and expressed in Escherichia coli system. After purification, lipase A activation was performed with the assistance of foldase lipase B. Enzyme assay revealed that the activated lipase A showed the optimal catalytic activity at 60ºC and pH 7. The effects of various metals on the activity were investigated and results demonstrated that most of the metals inhibited the activity. To further improve the catalytic outcome, two-phase reaction was studied, and n-hexane proved to be a good organic solvent for the combination system. Using the optimize conditions, (4R)-4-hydroxy-N, N-diphenyl-2-pentynamide with 94.5% ee value and 48.93% conversion ratio was achieved. Our investigation on this lipase reveals lipase A as a promising biocatalyst for producing chiral propargyl alcohol for preparation of novel himbacine analogs.

Enzymatic catalysis: An environmentally friendly method to enhance the transesterification of microalgal oil with fusel oil for production of fatty acid esters with potential application as biolubricants

This work aimed at the enzymatic synthesis of fatty acid esters with potential application as biolubricants from microalgae oil and fusel oil using Burkholderia cepacia lipase immobilized on Nb2O5. The lipid-bearing Dunaliella salina biomass was grown in bubble column photobioreactors, yielding 100 g of biomass and 20 g of oil containing high content of saturated (37.8%) and monounsaturated fatty acids (41.2%) and a low content of polyunsaturated fatty acids (20.9%). The reaction assays were carried out in 50 mL glass reactor containing 10 g of medium composed by microbial oil and fusel oil components (ethyl, butyl, or isoamyl alcohols) and fusel oil as acyl acceptors at a fixed molar ratio of 1:8 (oil to alcohol) using iso-octane as solvent. Batch runs were performed at 45 °C under constant magnetic stirring (150 rpm) for a maximum of 120 h. Macaw palm oil was used as a control under the same reaction conditions. The biocatalyst was found to be efficient in converting the microalgal oil fatty acids into their corresponding alkyl esters attained yields ranging from 70.8 to 89.0% at 120 h reaction, depending on size of the alcohol chain. The kinematic viscosities of the esters produced confirmed the high transesterification yields, considering the viscosity of microalgae oil decreased from 60.1 to a range of 8.9–11.3 m2 s−1 (40 °C) after the enzymatic modification.

Continuous flow reactor based with an immobilized biocatalyst for the continuous enzymatic transesterification of crude coconut oil.

Here, we have assessed the use of one packed bed or two packed bed reactors in series in which Burkholderia cepacia lipase (BCL) was immobilized on protic ionic liquid (PIL)-modified silica and used as a biocatalyst for the transesterification of crude coconut oil. Reaction parameters including volumetric flow, temperature, and molar ratio were evaluated. The conversion of transesterification reaction products (ethyl esters) was determined using gas chromatography and the quantities of intermediate products (diglyceride and monoglyceride [MG]) were assessed using high-performance liquid chromatography. Packed bed reactors in series produced ethyl esters with the greatest efficiency, achieving 65.27% conversion after 96H at a volumetric flow rate of 0.50mLMin-1 at 40°C and a 1:9 molar ratio of oil to ethanol. Further, within the first 24H of the reaction, increased MG (54.5%) production was observed. Molecular docking analyses were performed to evaluate the catalytic step of coconut oil transesterification in the presence of BCL. Molecular docking analysis showed that triglycerides have a higher affinity energy (-5.7kcalmol-1 ) than the smallest MG (-6.0kcalmol-1 ), therefore, BCL catalyzes the conversion of triglycerides rather than MG, which is consistent with experimental results.

Enantiodivergent Synthesis of Axially Chiral Biphenyls from σ‐Symmetric 1,1'‐Biphenyl‐2,6‐diol Derivatives by Single Lipase‐Catalyzed Acylative and Hydrolytic Desymmetrization

The enzymatic acylative desymmetrization of σ‐symmetric 2'‐halo‐1,1'‐biphenyl‐2,6‐diols was achieved for the first time using commercially available Burkholderia cepacia lipase immobilized on diatomaceous earth to give (S)‐mono esters. The hydrolytic desymmetrization of the corresponding diacetates was also achieved using the same lipase to give (R)‐mono esters. Our results, therefore, demonstrate that a single lipase can conduct the enantiodivergent synthesis of axially chiral biphenyl compounds in high chemical and optical yields.

Hydrophobic immobilization of Burkholderia cepacia lipase onto octyl-silica for synthesis of flavors esters

Burkholderia cepacia lipase (BCL) was immobilized onto silica modified with octyl groups (OS) and the biocatalyst (BCL-OS) was evaluated as its performance in the synthesis in organic medium (synthesis of flavor esters as a model). The maximum support loading was 0.375 g enzyme /g support , yielding a biocatalyst with an activity of 1197 U/g support at pH 7.0 and 50 °C in the hydrolysis of olive oil. The biocatalyst BCL-OS showed to be 9-fold more stable than the free lipase at 60°C in buffer solution (absence of substrates), with an increase of half-life from 16 to 144 h. The physical-chemical characterization of silica, octyl silica, and BCL-OS biocatalyst allowed confirming the immobilization of BCL onto the modified silica. The biocatalyst had an excellent performance in the synthesis of flavor esters, yielding more than 85% esterification yield (based on acid consumption) for acetic and butyric acids as acyl donors, and ethanol, butanol and hexanol as acyl acceptors. The biocatalyst could be recycled by ten 5 h-cycles of butyl butyrate syntheses at 37°C in heptane, retaining around 80% of its initial activity. Therefore, these results indicate that the BCL immobilized onto silica modified with octyl groups is a promising biocatalyst for application in organic syntheses.

Influence of additives in the transesterification of crude coconut oil catalyzed by Burkholderia cepacia lipase immobilized onto agro-industrial waste fibers

In this study was verified the influence of different additives in the transesterification reaction of the crude coconut oil catalyzed with lipase from Burkholderia cepacia immobilized by physical adsorption onto palm fiber originated from agroindustrial wastes. Reactions was performed under the conditions: molar ratio 1:7 (oil:alcohol), 10 % biocatalyst immobilized for 96 h at 40 °C in the presence in different concentrations of additives (water, tert -butanol, molecular sieve and protic ionic liquids). The results showed that the use of all the additives in this study did not increase conversion to ethyl esters. The maximum conversion in the absence of the additives was 72 %, in the presence of water 45 ± 2 %, molecular sieve 53 ± 2 % and tert -butanol 59 ± 2 %, respectively, all in the lowest concentrations. For ionic liquids, those with higher alkyl chains had the highest conversions, but lower than in their absence.

Surfactant Imprinting Hyperactivated Immobilized Lipase as Efficient Biocatalyst for Biodiesel Production from Waste Cooking Oil

Enzymatic production of biodiesel from waste cooking oil (WCO) could contribute to resolving the problems of energy demand and environment pollutions.In the present work, Burkholderia cepacia lipase (BCL) was activated by surfactant imprinting, and subsequently immobilized in magnetic cross-linked enzyme aggregates (mCLEAs) with hydroxyapatite coated magnetic nanoparticles (HAP-coated MNPs). The maximum hyperactivation of BCL mCLEAs was observed in the pretreatment of BCL with 0.1 mM Triton X-100. The optimized Triton-activated BCL mCLEAs was used as a highly active and robust biocatalyst for biodiesel production from WCO, exhibiting significant increase in biodiesel yield and tolerance to methanol. The results indicated that surfactant imprinting integrating mCLEAs could fix BCL in their active (open) form, experiencing a boost in activity and allowing biodiesel production performed in solvent without further addition of water. A maximal biodiesel yield of 98% was achieved under optimized conditions with molar ratio of methanol-to-WCO 7:1 in one-time addition in hexane at 40 °C. Therefore, the present study displays a versatile method for lipase immobilization and shows great practical latency in renewable biodiesel production.

SYNTHESIS OF ACILGLYCERIDES WITH HIGH NUTRITIONAL VALUE BY AN ENZYMATIC TWO-STEP PROCESS

Acylglycerides are esters formed from glycerol and fatty acids. The structure of these glycerides affects their digestion and absorption. In this work, the synthesis of acylglycerides with high nutritional value by an enzymatic process in two stages is presented. In the first stage, the esterification of glycerol and capric acid catalyzed by Lipozyme RM IM was carried out. In this reaction, 73% conversion of capric acid and 71% selectivity to 1,3-dicaprin was achieved. The diglyceride was separated and purified efficiently by a simple liquid-liquid extraction procedure. The dicaprin was esterified with palmitic acid using Burkholderia cepacia lipase immobilized on chitosan as the catalyst. In this second stage conversions of 1,3-dicaprin with values between 48 and 87% and with high selectivity towards the esterification of the sn-2 position were achieved. The reaction product had between 76 and 90% of acylglycerides with high nutritional value, depending on the reaction conditions.

Enhanced Activity of Immobilized Lipase by Phosphonium-Based Ionic Liquids Used in the Support Preparation and Immobilization Process

Biocatalysis has significant advantages over chemocatalysis in the context of sustainability since environmentally compatible catalysts (enzymes) and mild reaction conditions are used. However, enzymes are labile macromolecules, and strategies such as the addition of cosolvents or additives or their immobilization in solid supports are the target of intensive investigation to improve the catalytic performance and recyclability potential. In this work, we investigated the use of phosphonium-based ionic liquids (ILs) on the activity of immobilized Burkholderia cepacia lipase (BCL) by two approaches: (i) use of ILs to prepare silica used as support and (ii) use of ILs during the enzyme immobilization process. Several phosphonium-based ILs were investigated, allowing us to address the cation alkyl side chain and anion nature effects. The enzymatic performance depends on the IL employed to prepare silica, with a positive effect observed when employing ILs comprising cations with longer alkyl side chains and more hydrophobic anions. The best identified IL, namely, [P666(14)][NTf2], allows for a relative activity of 209.8% and immobilization yield of 77.3% and is capable of being recycled eight times (keeping more than 50% of the enzyme initial activity). When ILs are added during the BCL immobilization process, similar negative and beneficial effects are observed. With IL [P666(14)][NTf2], the immobilized biocatalyst has a relative activity of 322.7%, a total activity recovery yield of 91.1%, and can be recycled 17 times (down to 50% of the enzyme initial activity). Finally, both approaches were combined; i.e., IL [P666(14)][NTf2] was used both in the material preparation and immobilization process of the enzyme. This strategy allows for an increase in the relative activity up to 231%, an immobilization yield of 98%, and an increase of 9% in the enzyme relative activity. Although BCL activity is not significantly enhanced by this strategy, the combined use of the IL in silica preparation and during the enzyme immobilization process increased the recyclability potential of the immobilized biocatalyst material, capable of being recycled 26 times, while keeping more than 50% of the enzyme initial activity (equivalent to a half-life of 13 h). The results obtained in this work open the path to the efficient use of ILs, and particularly of the less explored phosphonium-based ILs, in the biocatalysis field.

Scaled-up biodiesel synthesis from Chinese Tallow Kernel oil catalyzed by Burkholderia cepacia lipase through ultrasonic assisted technology: A non-edible and alternative source of bio energy.

In East Asia, for thousands of years, the fruit of Chinese tallow tree (Sapium sebiferum) has been used for multiple purposes because of its chemical composition; the presence of high amounts of lipids is remarkable, showing potential to be used as substrate for biodiesel synthesis. Previously have been reported the use of alkaline and enzymatic catalysts, microwave technology and the use of ionic liquids as co-solvents with the lipids of this tree species to produce biodiesel. This study shows the results of the use of Burkholderia cepacia lipase as enzymatic catalyst for transesterification of Chinese Tallow Kernel oil (CTK), extracted from the fruit of Chinese tallow tree, into biodiesel, with the use of ultrasonic assisted technology and without the usage of solvents. The optimal operational parameters were determined and the reactions were developed in a batch reactor with the use of ultrasonic irradiation and emulsification to enhance the mass transfer. The scaled-up experiments, in an especially designed 3 L capacity reactor, showed promising results, obtaining 55.20% biodiesel and a kinematic viscosity of 10.31 mm2.s-1 in only 4 h, in comparison with previously published (in vitro) methods. The valorization of this non-edible source of oil represents an opportunity to use as an alternative source for bioenergy and also to tackle the uncontrolled expansion of this oleaginous tree species in some ecologically fragile ecosystems.

Surfactant Charge Modulates Structure and Stability of Lipase-Embedded Monolayers at Marine-Relevant Aerosol Surfaces.

Lipases, as well as other enzymes, are present and active within the sea surface microlayer (SSML). Upon bubble bursting, lipases partition into sea spray aerosol (SSA) along with surface-active molecules such as lipids. Lipases are likely to be embedded in the lipid monolayer at the SSA surface and thus have the potential to influence SSA interfacial structure and chemistry. Elucidating the structure of the lipid monolayer at SSA interfaces and how this structure is altered upon interaction with a protein system like lipase is of interest, given the importance of how aerosols interact with sunlight, influence cloud formation, and provide surfaces for chemical reactions. Herein, we report an integrated experimental and computational study of Burkholderia cepacia lipase (BCL) embedded in a lipid monolayer and highlight the important role of electrostatic, rather than hydrophobic, interactions as a driver for monolayer stability. Specifically, we combine Langmuir film experiments and molecular dynamics (MD) simulations to examine the detailed interactions between the zwitterionic dipalmitoylphosphatidylcholine (DPPC) monolayer and BCL. Upon insertion of BCL from the underlying subphase into the lipid monolayer, it is shown that BCL permeates and largely disorders the monolayer while strongly interacting with zwitterionic DPPC molecules, as experimentally observed by Langmuir adsorption curves and infrared reflectance absorbance spectroscopy. Explicitly solvated, all-atom MD is then used to provide insights into inter- and intramolecular interactions that drive these observations, with specific attention to the formation of salt bridges or ionic-bonding interactions. We show that after insertion into the DPPC monolayer, lipase is maintained at high surface pressures and in large BCL concentrations by forming a salt-bridge-stabilized lipase-DPPC complex. In comparison, when embedded in an anionic monolayer at low surface pressures, BCL preferentially forms intramolecular salt bridges, reducing its total favorable interactions with the surfactant and partitioning out of the monolayer shortly after injection. Overall, this study shows that the structure and dynamics of lipase-embedded SSA surfaces vary based on surface charge and pressure and that these variations have the potential to differentially modulate the properties of marine aerosols.

Chiral β-lactam-based integrin ligands through Lipase-catalysed kinetic resolution and their enantioselective receptor response

Obtainment and testing of pure enantiomers are of great importance for bioactive compounds, because of the assessed implications of enantioselectivity in receptor-mediated responses. Herein we evaluated the use of biocatalysis to obtain enantiomerically pure β-lactam intermediates further exploited in the synthesis of novel integrin ligands as single enantiomers. From a preliminary screening on a set of commercially available hydrolases, Burkholderia Cepacia Lipase (BCL) emerged as a suitable and highly performing enzyme for the kinetic resolution of a racemic azetidinone, key intermediate for the synthesis of novel agonists of integrins. Upon optimization of the biocatalytic protocol in terms of enzymes, acylating agents and procedures, the two β-lactam enantiomers were obtained in excellent enantiomeric excesses (94% and 98% ee). Synthetic elaborations on the separated enantiomers allowed the synthesis of four chiral β-lactams which were evaluated in cell adhesion assays on Jurkat cell line expressing α4β1 integrin, and K562 cell line expressing α5β1 integrin. Biological tests revealed that only (S)-enantiomers maintained the agonist activity of racemates with a nanomolar potency, and a specific enantio-recognition by integrin receptors was demonstrated.

Direct, selective and ultrasensitive electrochemical biosensing of methyl parathion in vegetables using Burkholderia cepacia lipase@MOF nanofibers-based biosensor

Methyl parathion is one of the most widely used pesticides in agricultural practices. It caused accumulation of acetylcholine and over-stimulation of receptors in synapses which eventually led to damage of the nervous system. Present study developed a direct, sensitive, rapid and reliable method for methyl parathion residues detection in vegetable samples. MOF nanofibers which demonstrated stable framework structure, good thermal/chemical stability, good electrochemical behavior, high porosity, surface area and pore volume was synthesized and used for fabrication of Burkholderia cepacia lipase (BCL)@MOF nanofibers biosensors. BCL@MOF nanofibers/chitosan/GCE biosensor demonstrated high sensitivity for methyl detection with a wide linear range (0.1–38 µM) and low limit of detection 0.067 µM. During the 3 weeks storage stability test at 4 °C, the fabricated biosensor demonstrated good reusability and excellent stability for methyl parathion detection with retainment of more than 80% of its initial response. When applied for detection of methyl parathion residues in vegetable samples, the BCL@MOF nanofibers/chitosan/GCE biosensors demonstrated good recovery rates.

MOF-Based Nanotubes to Hollow Nanospheres through Protein-Induced Soft-Templating Pathways.

A controllable and facile strategy is established for constructing metal–organic frameworks‐based (MOF‐based) hollow composites via a protein‐induced soft‐templating pathway. Using metal‐sodium deoxycholate hydrogel as soft‐template, nanotubes are gained while the protein is absent. With the presence of protein, hollow nanospheres structure are prepared by changing the amount of protein. To verify the universality of the proposed pathway, two kinds of proteins (Burkholderia cepacia lipase and penicillin G acylase) and three kinds of MOF (ZIF‐8, ZIF‐67, and Fe‐MOF) are adopted as model proteins and materials, and the obtained protein‐containing composites (named protein@H‐MOF) possess high bioactivity and stability. This proposed strategy provides a facile method for preparing MOF‐based composites under mild conditions, facilitating the applications of MOF in the fields of biocatalyst construction, biomolecule encapsulation, and drug delivery.

Molecular Basis of Aqueous-like Activity of Lipase Treated with Glucose-Headed Surfactant in Organic Solvent.

Lipases are useful as catalysts, particularly for the kinetic resolution of racemic alcohols and esters. However, their industrial applications are limited by their poor activities in organic media. We recently found that a lipoprotein lipase from Burkholderia species displays dramatically enhanced activity in organic solvent if the protein is coated with glucose-headed surfactant (GHS). Here we investigate the molecular basis of this enhanced enzymatic activity in organic solvent by performing molecular dynamics simulations on Burkholderia cepacia lipase as a model enzyme. Our simulation results indicate that the enhanced activity of lipase stems from a dual function of GHSs different in water and organic solvent. GHS molecules maintain the open conformation of lipase by providing lipid-like microenvironment surrounding the active site in water and stabilize its native active conformation by providing water-like microenvironment around the surface of the lipase in the organic solvent. Our data also suggest the role of organic cosolvent that can facilitate closed-to-open conformational changes during the freeze-drying process. The computational approach in this study lays its potential for guiding the design of more effective surfactant molecules to improve the activity of lipases in organic solvent.

Novel Sanger's Reagent-like Styrene Polymer for the Immobilization of Burkholderia cepacia Lipase.

Sanger's reaction, which was originally developed for amino acid detection, was utilized for enzyme immobilization. The newly synthesized polymer support, which was called polymer NO2-4-fluorostyrene-divinylbenzene (pNFD), was embedded with a Sanger's reagent-like functional group for immobilizing enzymes covalently under mild reaction conditions. Using Burkholderia cepacia lipase (BCL) as the target enzyme, the immobilization efficiency and activity of pNFD-BCL reached as high as 1.2 mg·g-1 and 33.21 U·g-1 (a specific activity of 27 675 U·g-1), respectively, realizing 90% activity recovery. It also improved the optimal reaction temperature of BCL from 40 to 65 °C, under which its full activity could be retained for 4 h. The new carrier also widened the pH-adaptive range of BCL as 6.5-10.0, allowing the lipase to operate normally in weak acid environment. Reusability of pNFD-BCL was significantly improved as almost no activity and/or enantioselectivity loss was observed in 200 h of triglyceride hydrolysis reaction and 17 batches of ( R, S)-1-phenylethanol resolution reaction.

Prediction of enantioselectivity of lipase catalyzed kinetic resolution using umbrella sampling

Enantiopure intermediates are preferred for drug synthesis in pharmaceutical industry. Lipases are widely used for chiral resolution of optically active compounds based on kinetic resolution. In the kinetic resolution, two enantiomers react with different rates in presence of a chiral catalyst or reagent, resulting in obtaining one enantiomer of great excess compared to other. The enantiomeric ratio (E) is closely related to the free energy difference (ΔΔG) of the activated state of substrate enantiomers. Molecular dynamic (MD) simulations with umbrella sampling technique can be used for estimation of activation free energy change of enantiomers. In this work, lipase-catalyzed transesterification of racemic alcohols with single and double hydroxyl groups have been studied. The umbrella sampling studies have been carried out for Candida rugosa lipase and Burkholderia cepacia lipase in n-hexane. The distance between serine residue in the catalytic triad and the ligand has been considered as a reaction coordinate and various conformations have been selected for MD simulation. The E value has been estimated based on free energy change from umbrella sampling. The estimated E values are in good agreement with experimental data. The work highlights changes in lipase conformation in n-hexane, ligand-protein interaction as well as free energy curve as a function of the reaction coordinate.

Enhancing enzyme activity and enantioselectivity of Burkholderia cepacia lipase via immobilization on melamine-glutaraldehyde dendrimer modified magnetic nanoparticles

Burkholderia cepacia lipase (BCL) was a potential catalyst in chiral resolution. Immobilization can be used to increase the enzyme activity and operational stability. In this work, novel melamine-glutaraldehyde dendrimer-like polymers were firstly grafted on the aminated magnetic nanoparticles to increase protein binding sites for the lipase to be immobilized. The dendrimer-polymer-modified nanoparticles could obviously protect the conformation of lipase and increase the contact chance between substrates and catalyzed center of lipase. The standard esterification reaction between lauric acid and 1-dodecanol was used to test the lipases activities. The activity of the obtained immobilized lipase was 58.0-fold than BCL powder, and 3.0-fold than the immobilized lipase without modified polymers. Then, the immobilized lipase was used to catalyze the chiral resolution reaction between 1-phenylethanol and vinyl acetate. Different organic solvents were selected for chiral resolution reaction. It was found out that hexane and heptane were the best reaction media. Thermal stability and organic solvent tolerance of the immobilized lipase were greatly improved than those of the lipase powder. The most significant discovery was the improvement in enzyme activity by bioimprinting. It could greatly shorten the reaction time from 3 h to 20 min. The conversion and substrate enantiomeric excess (ees) reached 49.7% and 98.8% in 20 min, respectively. The immobilized lipase could be recycled for 8 times without obvious loss in activity. The study provides a good strategy for preparation of efficient biocatalyst, showing great potential for future industrial application.

Hierarchical ZIF-8 toward Immobilizing Burkholderia cepacia Lipase for Application in Biodiesel Preparation.

A hierarchical mesoporous zeolitic imidazolate framework (ZIF-8) was processed based on cetyltrimethylammonium bromide (CTAB) as a morphological regulating agent and amino acid (l-histidine) as assisting template agent. Burkholderia cepacia lipase (BCL) was successfully immobilized by ZIF-8 as the carrier via an adsorption method (BCL-ZIF-8). The immobilized lipase (BCL) showed utmost activity recovery up to 1279%, a 12-fold boost in its free counterpart. BCL-ZIF-8 was used as a biocatalyst in the transesterification reaction for the production of biodiesel with 93.4% yield. There was no significant lowering of conversion yield relative to original activity for BCL-ZIF-8 when continuously reused for eight cycles. This work provides a new outlook for biotechnological importance by immobilizing lipase on the hybrid catalyst (ZIF-8) and opens the door for its uses in the industrial field.