FOOD RESEARCH INTERNATIONAL
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2026
·
IF 8.8
Daoming Li,Gaimiao Li,Jiandong Wang,Yuhao Li,Minguang Gao,Zhenbao Zhu,Duan Zhou
TDESs simultaneously enhance CALB activity, stability, and DAG selectivity. • ChCl:Gly:U (1,1:1) shows optimal catalytic performance. • Hydrogen-bond networks regulate CALB conformation and catalytic function. • DESs enable high-yield DAG synthesis with reduced by-product formation. Solvent engineering plays a crucial role in improving lipase performance in non-aqueous biocatalytic systems, particularly for the efficient and selective production of functional lipids. In this study, choline chloride-glycerol-urea ternary deep eutectic solvents (TDESs) were constructed to regulate the catalytic behavior of Candida antarctica lipase B (CALB), and the relationships between solvent physicochemical properties, enzyme conformation, and catalytic performance were systematically evaluated through enzymatic assays, fluorescence spectroscopy, and molecular dynamics simulations. Compared with binary DES and solvent free systems, TDESs exhibited lower water activity, tunable viscosity, and strengthened hydrogen-bonding networks, providing a more stable microenvironment for CALB. The optimal ChCl:Gly:U (1:1:1) system enhanced catalytic activity by 2.8-fold and retained over 80% residual activity after thermal treatment. Simulation results indicated reduced RMSD, RMSF, and SASA values, suggesting improved structural compactness. Subtle changes in interaction behavior within the catalytic triad (Ser105-His224-Asp187) were also observed, indicating localized modulation of active-site dynamics that may contribute to enhanced flexibility. In esterification, ChCl:Gly:U increased DAG content to 61% and reduced MAG content to <15% (vs. 45% DAG and 32% MAG in the solvent free system). In addition, the selectivity of DAG formation in TDES systems increased significantly from 57.66% to 73.61%. These findings demonstrate that rationally designed TDESs effectively enhance CALB activity, stability, and DAG selectivity, providing a feasible and sustainable strategy for enzymatic lipid modification. Download: Download high-res image (228KB) Download: Download full-size image
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高影响因子
JOURNAL OF CATALYSIS
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2025
·
IF 6.5
Mengyu Li,Wei Zhuang,Xia Meng,Wenxia Zhang,Keke Zhang,Zhenfu Wang
Metal-enzyme integrated catalysts combine the high selectivity of enzyme catalysts with the broad substrate spectrum of metal catalysts and offer tremendous possibilities for chemoenzymatic cascade processes. However, the rational design of microenvironments in nanostructures that make metals and enzymes compatible for efficient activity still remains challenging. In this report, a nanocomposite catalyst was fabricated by integrating ultrafine Pd nanoparticles (Pd NPs) and Candida antarctica lipase B (CALB) on the hydrophobic polydopamine-coated SiO 2 (SP) to enhance the compatibility of enzyme-metal catalysts. In detail, Pd NPs were in situ reduced by PDA on the SP surface, and the obtained SP-loaded Pd (Pd/SP) was hydrophobically modified by octadecyltrimethoxysilane for the subsequent adsorption immobilization of CALB to prepare the integrated catalyst, CALB/mPd/SP. The hydrophobic PDA coating on SiO 2 not only stabilized the loaded ultrasmall Pd nanoparticles but also facilitated the activation of the immobilized lipase, which helped to improve the cascade catalytic efficiency of CALB/mPd/SP. Afterward, CALB/mPd/SP was used in a one-pot dynamic kinetic resolution (DKR) reaction of α-phenylethylamine with high conversion (>99 %), selectivity (93.9 %), and ee p (>99 %). After 25 days of storage and 5 h of sonication, CALB/mPd/SP exhibited no significant reduction in the DKR catalytic activity. This study proposed a simple and sustainable method for the preparation of enzyme-metal cascade catalysts to enhance their stability and achieve significantly increased activity through interfacial microenvironmental modulation.
研究领域:
生物化学
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催化化学
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SEPARATION AND PURIFICATION TECHNOLOGY
·
2023
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IF 8.6
Jun Liang,Zhixi Zhang,Qiang Zhang,Yaqian Bian,Kunlong Yi,Wenbin Zhou,Weiyi Su,Yuqi Hu,Xiong Yu,Honghai Wang
Driven by global environmental awareness, biocatalysts are playing an increasingly important role in industrial chemistry . Enzymatic reactive distillation (ERD) combines the high efficiency of enzymatic reaction and distillation separation to reduce the limiting effects of reaction equilibrium, product inhibition, and the difficulty of product recovery due to its thermodynamic constraints. The lipase CALB was immobilized and coated on the θ-ring by sol-gel method. The synthesis of n-butyl acetate by transesterification reaction was selected as the model reaction in the ERD process, and a laboratory scale column was constructed. The Rate-based RadFrac model in Aspen Plus was used to describe the ERD. The results showed that the enzyme-loaded packing showed high long-term stability after 120 days in the reaction system. The deviation of the ERD experiment and simulation results was less than 10 %, which verifies the reliability of simulation. After the parameter sensitivity optimization on the simulation, compared with the batch reactor, the ERD process increased the reactant conversion from 60 % to 94.74 %. This study established a complete process from enzyme immobilization to ERD process, which provided a theoretical basis for the establishment and optimization of ERD process.
研究领域:
生物化学工程
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工业化学
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过程工程
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Catalysts
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2026
·Vol. 16
·期 5
·377
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IF 4.5
Sining Wei,Mahwish Aziz,Yilin Zhang,Jian Xiong,Cheng Cheng,Bin Wu
This study addressed the issue of insufficient activity in CALB lipase during the catalytic synthesis of key chiral intermediates for moxifloxacin. A structure-guided protein engineering strategy was employed to systematically modify its functional domains. Through molecular dynamics simulations of CALB-I189K, multiple regions exhibiting high conformational flexibility were preliminarily identified. Subsequently, by integrating 3D structural alignment with active site pocket distance analysis, the functionally most critical region (143–146) was selected. A site-directed saturation mutation library was constructed specifically targeting this region. Building upon the previously reported CALB-I189K, a mutant I189K/L144R/A146K was ultimately obtained through high-throughput screening combined with chiral HPLC validation. This mutant maintains excellent stereoselectivity (E = 206.52) while enhancing catalytic efficiency (kcat/Κm) to 273.73 min−1·mM−1, approximately 4.5-fold that of I189K. At a substrate concentration of 1 M, it achieves 50% conversion within 2.6 h, demonstrating kinetic resolution capabilities approaching industrial standards. Molecular simulation analysis indicates that the L144R and A146K mutations synergistically enhance catalytic performance primarily by optimizing spatial distances between catalytic residues. This study not only provides a high-performance catalyst for the efficient biosynthesis of moxifloxacin chiral intermediates but also offers new insights for enzyme rational design based on dynamic structural information.
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ACS Applied Nano Materials
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2023
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IF 5.9
Jiawei Lu,Ying-Nan Li,Youran Li,Huilin Zhu,Guiyang Shi
The reusability of immobilized lipases, including their hydrolysis activity, is greatly affected by lipase leakage, mass transfer channel blockage, and compound deposition on the outside surface. In this study, Candida antarctica lipase B (CALB) was immobilized on Fe3O4 nanoparticle-polyacrylonitrile (PAN) composite magnetic beads through phase inversion (L/FP). The hydrolysis activity of lipase was investigated using p-nitrophenyl palmitate (p-npp) as a model ester. To overcome the limitations of immobilized lipase in ester hydrolysis, three modification strategies were employed. First, lipase was covalently immobilized on Fe3O4 nanoparticles modified with methallyl (M9) and chloro (M13) silane modifiers, which exhibited faster immobilization with reduced lipase leaching and higher activity expression and stability. Second, poly(ethylene glycol) (PEG6000) and polylactic acid (PLA) were added to the precursors to physically modify the composite structures of L/FP, resulting in modified beads with larger specific surface area (15.26 m2·g–1) and smaller average pore size (19 nm). The modified beads, L/FP-9-M and L/FP-13-M, demonstrated superior reusability in long-term hydrolysis activity over 250 cycles. L/FP-9-M retained 63 and 34% of the initial activity at the 100th cycle and the 250th cycle, respectively. L/FP-13-M retained 55 and 36% of the initial activity at the 100th cycle and the 250th cycle, respectively. Furthermore, even after 250 repeated uses, the unit activities (U·gsupport–1) of both L/FP-9-M and L/FP-13-M remained higher than the initial activity of Novo 435.
研究领域:
生物化学
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材料科学
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酶工程
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Run Liu,Qi Zhou,Yi Zhang,Yuanzhi Xu,Zhonghui Liu,Kheng-Lim Goh,Vladimir Zivkovic,Mingming Zheng
Enzymatic synthesis of flavor esters is widely used in the food and flavor industries, but challenges remain in improving the catalytic efficiency and stability of biocatalysts. This study evaluates the performance of a novel biocatalyst, CALB@DMSN-C8, formed by immobilizing Candida antarctica lipase B (CALB) on hydrophobic dendritic mesoporous silica nanospheres (DMSN-C8), for synthesizing flavor esters. The CALB@DMSN-C8 catalyst achieves a caproic acid conversion rate of 98.5 ± 0.5% in just 30 min and demonstrates outstanding thermal stability, retaining a high conversion efficiency over 20 reuse cycles. To our knowledge, this study represents the most efficient synthesis of flavor esters, including ethyl valerate, ethyl caproate, ethyl heptanoate, and ethyl caprylate, compared to studies in the existing literature. Analysis of aroma characteristics and molecular docking simulations revealed the typical flavor profiles and synthesis mechanisms of various mellow esters. This study develops an innovative strategy by using self-made immobilized lipases to catalyze the production of flavor esters with potential applications in food and cosmetics.
研究领域:
食品科学
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酶工程
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生物催化
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PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
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2026
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IF 4.3
Shen Ji-Dong,Zhang Meng,Li Meng-Yang,Dong Yu-Qi,Liu Zhi-Qiang,Zheng Yu-Guo
Omega-3 polyunsaturated fatty acids (n-3 PUFAs) offer therapeutic benefits, including triglyceride reduction and management of cardiovascular and neurological disorders. This study investigates the use of fish oil processing by‑products, which consist mainly of fatty acid methyl and ethyl esters, as novel substrates for n‑3 PUFAs production. To overcome the low efficiency of conventional chemical methods and the vulnerability of free lipase to alcohol toxicity and poor reusability, we developed a stable immobilized lipase system. The target enzyme (CALB) was immobilized onto macroporous resin LXTE‑1000 via adsorption‑crosslinking, and optimal hydrolysis conditions were determined. The resulting immobilized enzyme (LXTE‑1000@CALB) showed markedly enhanced thermal and operational stability, retaining approximately 64.21% of its initial hydrolysis degree over ten consecutive batches. This work provides an efficient, green, and sustainable approach for valorizing fish oil by‑products and producing n‑3 PUFAs. Graphical abstract
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TALANTA
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2021
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IF 6.057
Zhentao Li,Qiaoyan Li,Yuanyuan Fu,Changjun Hu,Yikun Liu,Wen Li,Zilin Chen
Candida antarctica lipase B (CALB) is a natural biocatalyst with an intrinsically strong chiral environment and a high degree of enantio-selectivity, which is widely used in the separation of racemates . Here, a facile and efficient covalent immobilization approach was utilized to immobilize CALB onto the capillary inner wall as a novel chiral stationary phase to explore and broaden its application in the direct chiral separation by electrochromatography. The obtained CALB immobilized capillary column was characterized by scanning electron microscopy (SEM), fluorescence imaging and Fourier transform infrared spectroscopy (FT-IR). The enantioseparation property of the CALB immobilized capillary column was confirmed by direct chiral separation of several pairs of monoamine neurotransmitter enantiomers in OT-CEC mode. Outstanding enantioseparation performance for three types of monoamine neurotransmitter enantiomers including epinephrine, norepinephrine and phenylephrine was obtained by the CALB immobilized column. Thanks to the effectiveness of covalent bonding method and the intrinsic stability of CALB, the prepared CALB immobilized capillary columns were quite steady and reproducible. The relative standard deviations for retention times of the enantiomers were as follows: for intra-day (n = 5) runs (≤0.25%), inter-day (n = 3) runs (≤0.72%) and between-columns (n = 3) (≤2.42%). After 90 consecutive runs in CEC mode, the CALB immobilized column still exhibited desirable enantionseparation performance.
研究领域:
分析化学
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生物化学
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PubMed
高影响因子
Reaction Chemistry & Engineering
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2019
·Vol. 4
·期 6
·1136-1144
·
IF 3.935
Mengjie Wang,Meng Wang,Yun Zhu,Shengmiao Zhang,Jianding Chen
Enzyme functionalized porous beads were fabricated through covalently immobilizing Candida antarctica lipase B (CALB) onto high internal phase emulsion (HIPE) templated poly(glycidyl methacrylate) (PGMA) beads. These millimeter-sized beads were prepared by photopolymerizing the continuous phase of HIPE drops and then removing the dispersed phase. By covalently coupling the lysine residues of CALB to the epoxy groups from GMA, a series of enzyme immobilized polyHIPE beads were produced, with an enzyme loading of up to 0.18 g g−1 carrier. The resulting immobilized enzyme was highly stable to temperature and storage and could be recycled multiple times. The immobilized enzyme maintained 85% residual activity after 5 reaction cycles and 60% residual activity after 30 days of storage. The enzymatic esterification between hexanol and hexyl hexanoate had a conversion rate of up to 95%, showing that the enzyme immobilized beads were a good enzyme catalyst.
研究领域:
生物技术
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材料科学
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酶学
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高影响因子
EUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY
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2015
·Vol. 117
·期 10
·1636-1646
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IF 1.812
Lingyi Liu,Meirong Pang,Ying Zhang
AOB-w is a water-soluble antioxidant extracted from bamboo leaves, containing four flavone C-glucosides (orientin, isoorientin, vitexin, and isovitexin) as functional components. Flavone C-glucoside esters were firstly synthesized by three lipases (CALB, RM IM, and TL IM) in organic solvent system or solvent-free system. Results indicated that C12 fatty acid ester exhibited the optimal antioxidant capacity, CALB was the most efficient lipase in both systems, and higher conversion yields were achieved in solvent-free system. Highest conversion was obtained with 1% water content for CALB and 2% water content for RM IM and TL IM. All lipases displayed absolute 6″-regioselectivities with monoester. It is required to consider both of hydrophobicity and antioxidant capacity to prepare high-efficiency oil-soluble antioxidant. Regression equations were built up to illuminate the relationship between conversion yield of flavone C-glucoside and antioxidant capacity of its corresponding ester. Highest antioxidant capacity could be achieved when conversion yield was nearly 60% for flavone C-glucosides. Based on reaction parameters of flavone C-glucosides, high-efficiency eAOB-o was prepared, which could be expected as an excellent oil-soluble antioxidant in lipids. Practical applications: Water-soluble antioxidant of bamboo leaves (AOB-w) is a natural food antioxidant extracted from bamboo leaves, with four flavone C-glucosides (orientin, isoorientin, vitexin, and isovitexin) as functional components. This study aimed to provide a practical procedure to prepare high-efficiency lipid antioxidant by enzymatic acylation. Based on the optimal parameters from the synthesis of flavone C-glucoside esters, high-efficiency oil-soluble antioxidant of bamboo leaves (eAOB-o) was prepared. eAOB-o exhibited higher antioxidant capacity than cAOB-o, which has been approved as food antioxidants in China. High-efficiency acylation products were obtained by consideration both of lipid solubility and antioxidant capacity. Based on the rule, flavone C-glucoside ester and eAOB-o were prepared.
研究领域:
食品科学
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生物化学
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高影响因子
ACS Sustainable Chemistry & Engineering
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2025
·
IF 7.3
Chengkun Bai,Bingdi Wang,Chunying Lv,Zhengshun Jiang,Guolong Lu,Hang Sun,Zhenning Liu,Song Liang
Enzyme catalysis and photocatalysis utilizing solar energy are both promising pathways in sustainable chemistry. Drawing inspiration from the integrated enzyme-photocoupled systems in thylakoids, thylakoid-inspired microreactors (TIMs) were prepared using modified SiO2 nanoparticles as the building blocks, with g-C3N4-based photocatalysts encapsulated inside and CALB adsorbed on the surface. The thus designed TIMs result in exceptional catalytic efficiency in pyridine oxidation under visible light irradiation (>420 nm), achieving a rate 11.4 times greater than free enzymes and photocatalysts in a bulk solution. The increased contact area at the oil–water interface is the primary factor contributing to this enhancement, alongside the photocatalytic properties and enzyme loading. TIMs provide a robust platform for integrating functional components into a biomimetic, compartmentalized microreactor with spatially controlled organization and high-performance functionality.
研究领域:
生物化学
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材料科学
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可持续化学
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高影响因子
Molecular Catalysis
·
2025
·
IF 4.9
Huan-huan Fan,Wei Su,Jing-zhi Wu,Wen-long Liu,Chu-an Lin,Qing-yun Li,You-yan Liu,Ai-xing Tang
This study addresses challenges in the chemoenzymatic epoxidation of α-pinene, such as lipase deactivation, carrier decomposition in acidic environments, and product ring-opening side reactions, by developing a core-shell structured metal-organic framework (MOF)-based immobilized enzyme catalyst, CALB@ZIF-8@Zn-BTC. Candida antarctica lipase B (CALB) was encapsulated into ZIF-8 channels via in situ encapsulation. Trisodium citrate was introduced to accelerate the aggregation and co-precipitation of ZIF-8, while 1,3,5-benzenetricarboxylic acid (H₃BTC) coordinated with surface zinc ions of ZIF-8 to form a core-shell composite with enzyme protection and acid-buffering functionalities. Experimental results demonstrated that sodium citrate significantly enhanced acid scavenging capacity, suppressing acidic side reactions and achieving 96.6 % epoxidation selectivity. The Zn-BTC shell improved ZIF-8 stability in acidic systems, retaining over 60 % activity after 7 cycles. Under optimized conditions (3.35 mol/L ethyl acetate, 0.5 mol/L H₂O₂, 0.1 g sodium citrate), α-pinene conversion reached 83.1 %. Compared to the commercial catalyst Novozym435, CALB@ZIF-8@Zn-BTC exhibited superior catalytic efficiency, selectivity, and reusability. Structural characterization (FESEM, EDS, nitrogen adsorption) confirmed enhanced mass transfer and stability due to the composite’s unique architecture and uniformly distributed sodium citrate. This work provides an innovative solution for green and efficient α-pinene epoxidation and offers theoretical and practical insights for designing MOF-based catalysts in acidic environments.
研究领域:
催化
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绿色化学
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酶固定化
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金属有机框架材料
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有机合成
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化学反应工程
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高影响因子
危险品化学品经营许可证（带存储）