FIBERS AND POLYMERS
·
2018
·Vol. 19
·期 8
·1611-1617
·
IF 1.353
Wang Cuie,Wang Ning,Liu Xinhua,Wan Peng,He Xinwei,Shang Yongjia
We propose a green synthesis of benzopyran compound based on immobilized lipase. Firstly, by using a combined electrospun membrane and biocompatible chitosan, we show that the chitosan-functionalized electrospun PMA-co-PAA fibrous membrane can provide effective anchoring sites for immobilization of the Candida antarctica lipase B (CALB), and a maximum loading amount of CALB (152 mg/g) can be achieved. Then, the resulting membrane displays enhanced stability in catalyzing Knoevenagel condensation reaction of salicylaldehyde with active methylene compound in mixed aqueousorganic media. The catalytic activity and stability of the CALB on chitosan-modified fiber is better than its non-immobilized counterpart. Furthermore, this immobilize CALB catalyst still retained about 45 % of the initial activity after 5 cycles, which potentially had implications for the design of “green” enzyme catalyst for sustainable organic synthesis.
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Journal of Cleaner Production
·
2020
·
IF 7.246
Wei Zhang,Hairui Ji,Yanliang Song,Sen Ma,Wei Xiong,Changjing Chen,Biqiang Chen,Xu Zhang
To reduce the environment pollution from biolubricant synthesis, in this study, we developed a green and efficient strategy for the preparation of octylated branched biolubricant from waste cooking oil (WCO): (1) WCO was first hydrolyzed by a lipase ( Candida sp. 99–125 ) to produce unsaturated fatty acids (UFAs), the latter was subsequently concentrated by urea complexation; (2) Next, we synthesized new esters using the UFAs as substrate via esterification with 2-ethylhexanol employing lipase (Novozym 435) and the new esters products were further epoxidated; (3) After that, the epoxy group was attacked by a low-cost nucleophilic reagent, octanoic acid, to prepare octylated branched biolubricant using an recyclable ionic liquid , [HMIm][PF 6 ], as catalyst. After three recycling times, [HMIm][PF 6 ] remained catalytic activity with a conversion yield of 81.22% and selectivity of 82.10%. The obtained product showed excellent lubricant properties, such as low pour point of −61 °C, high viscosity index of 149 and high thermal-oxidation stability (onset temperature of 326.35 °C and oxidative onset temperature of 312.06 °C). The high frequency reciprocating rig (HFRR) measurement demonstrated that lubrication performance (friction coefficient of 0.09 and wear spot diameter of 203 μm) was much better than that of mineral based lubricant with the same viscosity grade. Therefore, this study provided a critical synthetic strategy for preparing high-performance biolubricants from WCO.
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Journal of the Taiwan Institute of Chemical Engineers
·
2019
·
IF 3.834
Wenjing Cheng,Yun Li,Xiaoying Li,Wenjing Bai,Yunxiao Liang
A monolithic macroporous silica (MMS) was prepared by using a pillared epoxy resin macroporous polymer as template. Polydopamine (PDA) as a spacer-arm polymer was modified on the surface of MMS for immobilization of Candida rugosa lipase (CRL), aiming to improve the performance of CRL and overcome the limitations of the immobilized enzymes on mesoporous materials and fine particles. The immobilization conditions and the properties of the immobilized CRL were investigated. The as-prepared monolithic macroporous materials were characterized by SEM, TEM, MIP, BET, FT-IR and TG-DTA. The results revealed that MMS possessed two sets of well-defined and uniform continuous macroporous channels (pore size of 300–500 nm), high mechanical strength, large pore volume and specific surface area, and that the PDA/MMS macroporous composite was constructed by PDA/SiO 2 /PDA nano-film with a thickness of about 50 nm. Because of its surface chemical properties and unique macropore structure, the PDA/MMS composite provided a simple, cost-effective way to enhance the catalytic activity of directly immobilized enzyme. The activity recovery of CRL–PDA/MMS was 201%, and the stability of the immobilized lipase was remarkably improved. Moreover, this biocatalyst can be easily recovered from the reaction system without centrifugation or filtration.
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ACS Sustainable Chemistry & Engineering
·
2019
·
IF 6.97
Hongbo Suo,Lili Xu,Chao Xu,Xiang Qiu,Hongyue Chen,He Huang,Yi Hu
In this study, a novel enzyme immobilization method was developed to enhance the catalytic stability of enzymes. In this strategy, ionic liquid (IL) modified magnetic chitosan (MCS) composites were used as supports for lipase adsorption and graphene oxide (GO) was employed as shell coating for the first time. The modifier used was imidazolium-based IL with a side alkyl chain which was composed of 8 −CH2 and a terminal hydroxyl group. The prepared supports IL-MCS, immobilized lipase PPL-IL-MCS, and GO/PPL-IL-MCS were well characterized. The GO shielding lipase GO/PPL-IL-MCS maintained high activity (2468 U/g), which was 6.72-fold of free lipase. In addition, the pH and temperature effect on lipase activity were investigated. The thermal stability, denaturants stability, storage stibility, and reusing stability were also studied. Compared to PPL-IL-MCS, the stabilities of GO/PPL-IL-MCS were all enhanced while keeping high activity. For example, after 10 cycles of reuse, the residual activity of GO/PPL-IL-MCS was 92.1%, which was higher than that of 88.4% for PPL-IL-MCS. Furthermore, the apparent Km of PPL-IL-MCS and GO/PPL-IL-MCS was 5.7 and 8.8 mg/mL, respectively, which were both lower than that of PPL-MCS (17.1 mg/mL). Circular dichroism (CD) was used to analyze the secondary structure of lipase to explain the mechanism of stable enhancement of immobilized enzyme. This work demonstrated that GO was used as a shell coating for the first time to improve the lipase stability. This immobilization method provides a reference for the immobilization of other kinds of enzymes.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Yong Wang,Ziyuan Wang,Huifen Yu,Haidong Teng,Jianping Wu,Jiaqi Xu,Lirong Yang
Lipase from Rhizopus oryzae (ROL) exhibits remarkable sn-1,3 stereoselectivity and catalytic activity, but its poor thermostability limits its applications in the production of 1,3-dioleoyl-2-palmitoyl glycerol (OPO, a high-quality substitute for human milk fat). In this work, a semirational method was proposed to engineer the thermostability and catalytic activity of 4M (ROL mutant in our previous study). First, a computer-aided design is performed using 4M as a template, and N-glycosylation mutants are then recombinantly expressed and screened in Pichia pastoris, the optimal mutant N227 exhibited a half-life of 298.8 h at 45 °C, which is 7.23-folds longer than that of 4M. Its catalytic activity also reached 1043.80 ± 61.98 U/mg, representing a 29.2% increase compared to 4M (808.02 ± 47.02 U/mg). Molecular dynamics simulations of N227 suggested that the introduction of glycan enhanced the protein rigidity, while the strong hydrogen bonds formed between the glycan and the protein stabilized the lipase structure, thereby improving its thermostability. The acidolysis reaction between oleic acid (OA) and glycerol tripalmitate (PPP) was successfully carried out using immobilized N227, achieving a molar conversion rate of 90.2% for PPP. This engineering strategy guides the modification of lipases, while the glycomutants obtained in this study have potential applications in the biosynthesis of OPO.
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高影响因子
PROCESS BIOCHEMISTRY
·
2020
·
IF 2.952
Jian Ou,Xin Yuan,Yu Liu,Panliang Zhang,Weifeng Xu,Kewen Tang
Enzyme immobilization in MOFs offers retained enzyme integrity and activity, enhanced stability, and reduced leaching. In this work, Pseudomonas cepacia lipase (PCL) was successfully immobilized into the Zeolitic imidazolate framework-8 (ZIF-8) by physical adsorption. The amounts of PCL in the immobilized enzyme (PCL@ZIF-8) was determined to be 16.7 % (200.5 mg PCL/g ZIF-8). Furthermore, the immobilized enzyme was applied as efficient bio-catalyst for enantioselective hydrolysis of 2-phenylpropionic acid (2-PPA) ester enantiomers and enantioselective transesterification of 1-phenylethanol enantiomers. The enzymatic activity of the immobilized enzyme was three times more than that of free PCL in enantioselective hydrolysis system, and the enantiomeric excess was maintained above 99 %. There was no significant difference in enzyme activity between immobilized PCL and free PCL in transesterification system. In addition, the immobilized enzyme showed good reusability in both hydrolysis (30.57 % of initial activity, 4 cycle) and transesterification reaction systems (64.38 % of initial activity, 6 cycle).
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BIORESOURCE TECHNOLOGY
·
2026
·
IF 8.2
Haoyu Chai,Xin Li,Yu-Cai He,Cuiluan Ma
Recombinant E. coli ADH6–GDH carrying ADH6 and GDH was constructed for bioreduction. • ADH6–GDH whole-cell fully converted 40 mM syringaldehyde to syringyl alcohol. • Substrate spectrum was explored using lignin-derived aldehydes via bioreduction. • Lipase Novozym 435 enabled the conversion of syringyl alcohol to syringyl acetate. • Tandem biocatalysis with reductase and lipase was used to synthesize aromatic esters. Efficient valorization of lignin-derived aromatics remains a key challenge in sustainable chemistry. Biobased aromatic alcohols and esters are important value-added compounds widely used in fragrances, pharmaceutical intermediates, fine chemicals, and functional materials. In this study, a whole-cell biocatalytic system co-expressing alcohol dehydrogenase (ADH6) from Saccharomyces cerevisiae and glucose dehydrogenase (GDH) was developed for the efficient conversion of syringaldehyde to syringyl alcohol. The GDH-based cofactor regeneration module enabled in situ reduced nicotinamide adenine dinucleotide phosphate (NADPH) supply, thereby improving catalytic efficiency while reducing the dependence on external cofactors. Under optimized conditions, the system achieved a high analytical yield of 98.3% at 40 mM syringaldehyde in an aqueous system, representing, to the best of our knowledge, the highest substrate concentration reported so far for this biocatalytic conversion. The substrate spectrum was further evaluated using representative lignin-derived aromatic aldehydes, including cinnamaldehyde, p -anisaldehyde, vanillin, and 3,4-dimethoxybenzaldehyde. These substrates were converted into cinnamyl alcohol, 4-methoxybenzyl alcohol, vanillyl alcohol, and 3,4-dimethoxybenzyl alcohol, respectively. Cinnamaldehyde, p -anisaldehyde, and 3,4-dimethoxybenzaldehyde afforded the corresponding alcohols with analytical yields of 99%, whereas vanillin gave vanillyl alcohol with an analytical yield of 86% after 4 h. Molecular docking indicated that these substrates could bind within the ADH6 active pocket through similar orientations, supported by hydrogen-bonding and hydrophobic interactions, providing structural explanation for the observed substrate adaptability. In addition, coupling with immobilized lipase Novozym 435 enabled the conversion of syringyl alcohol into syringyl acetate via transesterification. This work established an efficient tandem biocatalysis with reductase and lipase for upgrading of lignin-derived aromatic compounds. Download: Download high-res image (100KB) Download: Download full-size image
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产品关联
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高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2026
·
IF 6.7
Qiongya Lu,Jingcheng Hu,Xing Ni,Xue Zhang,Jiaojiao Xia,Yani Zhao,Rubing Huang,Bin Zou
In this study, we successfully synthesized two immobilized enzyme carrier: hollow mesoporous carbon spheres (HMCS) and mesoporous carbon spheres (MCS). The carriers were thoroughly characterized by using a range of techniques, including scanning electron microscopy, water contact angle measurement, and BET surface area analysis. HMCS exhibited a unique hollow structure, moderate hydrophobicity, and a larger specific surface area. The high-performance immobilized lipase, CRL@HMCS was prepared by using HMCS as the carrier. The basic enzymatic properties and reaction kinetic parameters of CRL@HMCS were studied. CRL@HMCS exhibited significantly higher catalytic efficiency, with a Vmax approximately 2.7-fold greater than that of the free CRL. Excellent operational stability was showed by retaining 64.14% of its relative activity after being reused 7 times and 84.05% after 22 days of storage. Under optimal conditions (45 °C, 200 rpm, 5 h), CRL@HMCS achieved a 70.4% yield in geranyl acetate synthesis, significantly higher than the free enzyme (48.65%).
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高影响因子
LANGMUIR
·
2026
·
IF 4.4
Ting Xu,Baoxin Yang,Ranran Guo,Linling Yu,Xiaoyan Dong,Yan Sun
Bacillus thermocatenulatus lipase 2 (BTL2) is a highly versatile enzyme for catalyzing the hydrolysis and synthesis of various esters, but the practical application of the enzyme is limited by its poor operational stability and difficulty in recovery. To address these limitations, we have herein proposed a dual fusion strategy that combines the chaperone-like protein α-synuclein (αS) at the C-terminus and silica-binding peptide (SiBP) at the N-terminus, making a fusion enzyme (SiBP-BTL2-αS) for enhanced enzymatic performance and site-specific immobilization on mesoporous silica nanoparticles (MSNs) for repeated use. The catalytic activity of the enzymes was evaluated using a colorimetric p-nitrophenyl palmitate (pNPP) assay at 30 °C in 50 mM HEPES buffer (pH 8.0), and relative activity was expressed as the ratio to the wild-type BTL2. It was found that free SiBP-BTL2-αS showed a 1.8-fold higher activity than BTL2 due to the chaperone effect of αS, and immobilization on MSNs brought out a further 1.4-fold increase in activity at an enzyme loading of 194 mg/g. Thus, SiBP-BTL2-αS@MSNs presented 3.3-fold higher activity than BTL2. Moreover, SiBP-BTL2-αS@MSNs exhibited significantly improved thermostability and broad pH tolerance over the free counterpart and BTL2. In repeated uses, SiBP-BTL2-αS@MSNs retained 82.1% of its initial activity after seven consecutive reaction cycles. In the synthesis of vitamin E succinate, SiBP-BTL2-αS@MSNs showed 32% and 78% higher yields over SiBP-BTL2-αS and BTL2, respectively, verifying the superiority of SiBP-BTL2-αS@MSNs in enzymatic catalysis. This work not only offers a highly efficient, robust, and recyclable enzyme preparation, but also provides a promising way to design immobilized lipase with hyperactivation behavior.
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高影响因子
BIORESOURCE TECHNOLOGY
·
2026
·
IF 8.2
Meijun Du,Jun Jin,Cong Sun,Abdelaziz Elbarbary,Wei Wei,Hui Zhang,Qingzhe Jin,Xingguo Wang,Zhe Cui
Immobilized CRL enables selective esterification of CLA in a non-aqueous system. • Hydrophobic acyl acceptors enhance esterification efficiency and selectivity. • n -Octanol is identified as the optimal acyl acceptor. • Two-step esterification yields high-purity c9,t11- and t10,c12-CLA. An immobilized Candida rugosa lipase on mesoporous silica functionalized with C18 alkyl chains (MS-C18) was employed to construct a non-aqueous selective esterification system for the resolution of conjugated linoleic acid (CLA) isomers. The effects of acyl acceptors with different hydrophobicities on esterification performance and isomer selectivity were investigated in isooctane. Under controlled reaction conditions (CLA-to-alcohol molar ratio 1:1, 40 °C), increasing acyl acceptor hydrophobicity enhanced both the esterification rate and the differentiation of reaction rates between c9,t11-CLA and t10,c12-CLA, with n -octanol identified as the optimal acyl acceptor. A two-step esterification strategy, involving initial separation of c9,t11-rich ester and t10,c12-rich free fatty acid fractions from commercial CLA mixtures, followed by secondary esterification of the respective free fatty acid substrates, enabled simultaneous preparation of both isomers with high relative purity (≥90%) and total recovery of 38.1%-52.4%. Kinetic analysis indicated that the reaction follows an alcohol-inhibited Ping-Pong Bi-Bi mechanism, and the observed selectivity is associated with differential substrate affinity and catalytic efficiency of CRL toward the two CLA isomers. The use of more hydrophobic acyl acceptors likely reduces substrate inhibition and enhances the differentiation of reaction rates between isomers, thereby improving separation efficiency. This study demonstrates that tuning acyl acceptor hydrophobicity provides an effective strategy to regulate selective esterification in non-aqueous systems, offering a feasible and scalable approach for high-purity CLA isomer preparation. Download: Download high-res image (140KB) Download: Download full-size image
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高影响因子
3 Biotech
·
2026
·Vol. 16
·期 6
·219
·
IF 3.1
Wang Yuanshan,Liu Ping,Yang Xiaobo,Xu Jiabao,Qin Haibin,Weng Chunyue,Zheng Yuguo
A robust and reusable Rhizomucor miehei lipase (RML) biocatalyst was prepared by integrating bio-imprinting with resin immobilization. RML was co-imprinted with lauric acid and polyethylene glycol (PEG600) followed by immobilization on an amine-functionalized epoxy resin (ES-103B), resulting in AE@RML-LA+PEG600. After immobilization process optimization, the AE@RML-LA + PEG 600 was verified by CD, SEM, FTIR, and XRD analysis. AE@RML-LA+PEG600 exhibited improved thermal, pH, and organic solvent stability compared with free RML. Under optimized conditions (10 h, 62 °C, palmitic-acid-enriched palm stearin/oleic acid = 1:10 mol/mol, 10 wt% enzyme), AE@RML-LA+PEG600 achieved a maximum OPO content of 45.80%, and the proportion of palmitic acid at the sn-2 position relative to total palmitic acid ((sn-2, P)/P) was 54.59%, which was higher than that of the unimprinted enzyme (35.52% OPO). After five cycles, AE@RML-LA+PEG600 maintained 69.52% of its original catalytic activity, indicating its excellent operational stability. This work provides a promising imprinting-immobilization strategy for preparing durable lipases for industrial OPO production.
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高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2022
·Vol. 45
·期 4
·749-759
·
IF 3.434
Zhang Xiao-guang,Xue Yu,Lu Ze-ping,Xu Hua-jin,Hu Yi
In this study, a series of proline ionic liquids with different lengths of hydrophobic alkyl on the side chain were used to modify the Candida Antarctic lipase B (CALB). The catalytic activity, thermal stability and tolerance to methanol and DMSO of the modified enzyme were all improved simultaneously. The optimum temperature changed from 55 to 60 ℃. The hydrophobicity and anion type of the modifier have important influence on the catalytic performance of CALB. CALB modified by [ProC 12 ][H 2 PO 4 ] has a better effect. Under the optimal conditions, its hydrolysis activity was 3.0 times than that of the native enzyme, the catalytic efficiency Kcat/Km improved 2.8 times in aqueous phase, and the tolerance to organic solvent with strong polarity (50% methanol 2 h) was increased by 6.8 times. Fluorescence spectra and circular dichroism (CD) spectroscopy showed that the introduction of ionic liquids changed the microenvironment near the fluorophores of the enzyme protein, the α-helix decreased and β-sheet increased in the secondary structure of the modified enzymes. The root mean square deviation (RMSD), residue root mean square fluctuation (RMSF), radius of gyration (Rg), and solution accessible surface area (SASA) of [ProC 2 ][Br]-CALB, [ProC 12 ][Br]-CALB and native CALB were obtained for comparison by molecular dynamics simulation. The results of dynamics simulation were in good agreement with enzymology experiment. The introduction of ionic liquids can keep CALB in a better active conformation, and proline ionic liquids with long hydrophobic chains can significantly improve the surface hydrophobicity and overall rigidity of CALB. This research offers a new idea for rapid screening of efficient modifiers and provision of enzymes with high stability and activity for industrial application.
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高影响因子
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