FIBERS AND POLYMERS
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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.
研究领域:
生物化学
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绿色化学
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材料科学
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DOI
高影响因子
PROCESS BIOCHEMISTRY
·
2024
·
IF 3.7
Shihao Liang,Wei Zhuang,Zhenfu Wang
In this work, isocyanate-based strategy was developed for efficient immobilization of Candida antarctica lipase B (CALB). Of 41 commercial resins, EP109 was chosen and hydrophobized with hydroxyl-terminated polydimethylsiloxane (HTPDMS) for tert-butyl isocyanate (tBIC)-based CALB immobilization due to high loading amount and specific activity. The tBIC-based strategy exhibited shorter immobilization time, higher loading and activity than adsorption strategy by hydrophobic interaction. When compared with free CALB, EP109-HTPDMS-tBIC@CALB owned improved stability (against acid, alkali, and heat) and storability. Subsequently, EP109-HTPDMS-tBIC@CALB was employed in the bio-synthesis of 1,3-diacylglycerol in a solvent-free system with a yield of 76.31%, which was higher than commercial Novozym 435 (53.93%). EP109-HTPDMS-tBIC@CALB exhibited 76.34% of residual activity after 20 repeat uses.
研究领域:
生物技术
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酶工程
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化学工程
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生物化学
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高影响因子
RSC Advances
·
2017
·Vol. 7
·期 21
·12518-12523
·
IF 3.108
Pengfei Zhou,Xuping Wang,Bo Yang,Frank Hollmann,Yonghua Wang
Epoxides are important synthetic intermediates for the synthesis of a broad range of industrial products. This study presents a promising solution to the current limitation of enzyme instability. By using simple deep eutectic solvents such as choline chloride/sorbitol, significant stabilization of the biocatalyst has been achieved leading to more robust reactions while using environmentally more acceptable solvents as compared to ionic liquids.
研究领域:
生物化学
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绿色化学
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高影响因子
BIOTECHNOLOGY PROGRESS
·
2021
·Vol. 38
·期 1
·e3218
·
IF 2.681
Xiao-Jian Zhang,Feng-Yu Qi,Jia-Mei Qi,Fei Yang,Jiang-Wei Shen,Xue Cai,Zhi-Qiang Liu,Yu-Guo Zheng
The Candida antarctica lipase B (CALB) was embedded in the metal-organic framework, zeolitic imidazolate framework-8 (ZIF-8), and applied in the enzymatic synthesis of L -ascorbic acid palmitate (ASP) for the first time. The obtained CALB@ZIF-8 achieved the enzyme loading of 80 mg g −1 with 11.3 U g −1 (dry weight) unit activity, 59.8% activity recovery, and 92.7% immobilization yield. Under the optimal condition, ASP was synthesized with over 75.9% conversion of L -ascorbic acid in a 10-batch reaction. Continuous synthesis of ASP was subsequently performed in a packed bed bioreactor with an outstanding average space–time yield of 58.1 g L −1 h −1 , which was higher than ever reported continuous ASP biosynthesis reactions.
研究领域:
生物化学
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酶工程
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材料科学
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PubMed
高影响因子
Bioresources and Bioprocessing
·
2025
·Vol. 12
·期 1
·1-16
·
IF 5.1
Li Yuhan,Wu Guowei,Liu Zeqing,Dai Lingmei,Liu Dehua,Du Wei
Mesoporous MIL-88A (Meso-MIL-88A) shows significant potential as an effective carrier for immobilizing large molecules such as lipases. This study investigates Meso-MIL-88A immobilized lipase for the catalysis of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) incorporation into soybean phosphatidylcholine. Phosphatidyl-EPA and phosphatidyl-DHA are known for their superior health benefits and have garnered significant global attention. Firstly, we developed a novel and green water-washing strategy to synthesize Meso-MIL-88A, demonstrating its significant potential as an effective carrier for immobilizing large molecules such as lipases. Nitrogen adsorption/desorption and XPS analyses revealed that water as the eluent yielded larger average pore diameters and lower sulfur residue content compared to ethanol. This optimized Meso-MIL-88A carrier was used to immobilize Candida antarctica lipase B (CalB@Meso-MIL-88A), which was applied to the production of phosphatidyl-EPA and phosphatidyl-DHA. The immobilized CalB@Meso-MIL-88A exhibited exceptional catalytic efficiency, achieving an unprecedented sn -1 positional incorporation rate of 86.8% (44.2% EPA, 42.7% DHA) with 90% EPA/DHA-ethyl ester (EE) donors, while 97% DHA-EE resulted in a record-high 90.1% DHA incorporation at the sn -1 position. Kinetic studies and molecular docking simulations indicated a higher substrate affinity for DHA-EE, attributed to enhanced transfer efficiency of EPA/DHA-EE in organic solvent system. This study presents the first demonstration of the potential of Meso-MIL-88A for industrial lipase immobilization via this optimized route, offering an eco-friendly and highly efficient catalytic application for nutraceutical synthesis. Graphical abstract
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高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2017
·
IF 2.502
Kun Zhang,Zhiyou Pan,Zhoujian Diao,Shuli Liang,Shuangyan Han,Suiping Zheng,Ying Lin
Kinetic resolution of sec -alcohols is a green process with biocatalyst. Candida antarctica lipase B (CALB) displayed on Pichia pastoris cell-surface (Pp-CALB) was characterized in kinetic resolution of sec -alcohols with different structures. The reaction parameters including acyl donors, molar ratio of substrates, solvents and temperatures were examined with 2-octanol as model substrate. 47.4% molar conversion of 2-octanol and 99.7% ee p were obtained after a 5 h reaction with Pp-CALB, and 90% of its original activity still remained after being reused for 10 cycles. Pp-CALB was then used to several sec -alcohols and it showed great enzymatic activity and enantioselectivity to all tested sec -alcohols, more than 93.1% of ee p . The enantioselective characteristics of Pp-CALB catalysed sec -alcohols with different structures were compared with Novozyme 435 which was almost the same. Solvent free system as one way of green chemistry was applied to Pp-CALB and Pp-CALB showed great catalytic activity and enantioselectivity. Pp-CALB was potential biocatalyst of high enzymatic activity and enantioselectivity using in resolution of sec -alcohols.
研究领域:
生物化学
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酶工程
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绿色化学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2026
·
IF 10.4
Jiawei Zheng,Xuan Hu,Jiufang Chen,Fengxia Lan,Li Xu,Zeping Lu,Binbin Nian,Nanjing Zhong
CALB@GA-NH 2 -UiO-66 exhibited 97.31% oleic acid conversion and 77.60% DAG content. • CALB@GA-NH 2 -UiO-66 retained full initial activity after 11 reuse cycles. • The immobilized CALB exhibited a single conformational basin. • Covalent enzyme–carrier conjugates were formed via Schiff base linkages. • The immobilization moderately restrained the flexibility of α-helix 5 of CALB. This study aimed to develop a robust biocatalyst for diacylglycerol (DAG) synthesis. Candida antarctica lipase B (CALB) was immobilized on NH₂-UiO-66 via glutaraldehyde (GA) cross-linking. The cross-linked CALB exhibited significantly better performance than the adsorbed counterpart. Oleic acid conversion reached 97.31% with DAG content up to 77.60%. Moreover, the immobilized CALB retained its full initial activity after 11 reuse cycles. The temperature dependence of oleic acid conversion followed the equation ln V₀ = 8.86–4.67 × 10 3 / T , with an activation energy of 38.83 kJ·Mol −1 . Molecular dynamics simulations revealed that the flexibility of α-helix 5 in CALB@GA-NH₂-UiO-66 is moderately restrained, and the immobilized CALB exhibits a single conformational basin. Additionally, immobilization induces slight contraction of the CALB active site pocket, and reduced length and increased bottleneck radius of the tunnel. This study provides molecular insights into the structure–stability relationship of immobilized enzymes, guiding the rational design of robust biocomposites We develop a robust biocatalyst via glutaraldehyde cross-linking of CALB onto NH₂-UiO-66 for DAG production by esterification. Download: Download high-res image (210KB) Download: Download full-size image
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ORGANIC & BIOMOLECULAR CHEMISTRY
·
2013
·Vol. 11
·期 41
·7192-7198
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IF 3.568
Ru Jia,Yi Hu,Luo Liu,Ling Jiang,He Huang
Various imidazolium-functional ionic liquids (ILs) composed of different cations and anions were grafted onto Candida antarctica lipase B (CALB) through lysine coupling, and 4–6 of the 9 primary amino groups of the lysine residues were modified. The catalytic activity and stability were investigated in a p-nitrophenyl palmitate hydrolysis reaction. After modification, CALB was activated and achieved a high catalytic efficiency in the aqueous phase (e.g., [HOOCMMIm][Cl] modification showed a 1.5-fold increase in the catalytic efficiency). The thermostability and organic solvent tolerance were significantly increased with the ILs modification (e.g., [HOOCBMIm][Cl] owned kosmotropic cation and chaotropic anion modification showed a 7-fold thermostability increase at 70 °C, 1-fold increase in 50% aqueous dimethylformamide and 5-fold increase in 50% aqueous methanol). Conformation changes were confirmed by fluorescence spectroscopy, circular dichroism spectroscopy and attenuated total reflection Fourier transform infrared spectroscopy.
研究领域:
生物化学
·
酶工程
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产品关联
DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2023
·
IF 3.4
Liwen Shu,Xin Zheng,Shuhua Qi,Shuizhi Lin,Yinghua Lu,Chuanyi Yao,Xueping Ling
Docosahexaenoic acid (DHA) enriched with phospholipids (PLs) (DHA-PLs) is a type of structured PL with good physicochemical and nutritional properties. Compared to PLs and DHA, DHA-PLs has higher bioavailability and structural stability and many nutritional benefits. To improve the enzymatic synthesis of DHA-PLs, this study investigated the preparation of phosphatidylcholine (PC) enriched with DHA (DHA-PC) via enzymatic transesterification of algal oil, which is rich in DHA-triglycerides, using immobilized Candida antarctica lipase B (CALB). The optimized reaction system incorporated 31.2% DHA into the acyl chain of PC and converted 43.6% PC to DHA-PC within 72 h at 50 °C, 1:8 PC: algal oil mass ratio, 25% enzyme load (based on total substrate mass), and 0.02 g/mL molecular sieve concentration. Consequently, the side reactions of PC hydrolysis were effectively suppressed and products with high PC content (74.8%) were produced. Molecular structure analysis showed that exogenous DHA was specifically incorporated into the sn-1 site of the PC by immobilized CALB. Furthermore, the evaluation of reusability with eight cycles showed that the immobilized CALB had good operational stability in the present reaction system. Collectively, this study demonstrated the applicability of immobilized CALB as a biocatalyst for synthesizing DHA-PC and provided an improved enzyme-catalyzed method for future DHA-PL synthesis.
研究领域:
食品科学
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生物化学
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PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2024
·
IF 3.5
Liu Jiale,Zhao Shufan,Wei Wan,Yu Shupeng,Wang Zhao,Zheng Jianyong
Lipase is one of the most widely studied and applied biocatalysts. Due to the high enzyme leakage rate of the immobilization method of physical adsorption, we propose a new lipase immobilization method, based on the combination of macroporous resin adsorption and organic polymer coating. The immobilized Candida antarctica lipase B (CALB@resin-CAB) was prepared by combining the macroporous resin adsorption with cellulose acetate butyrate coating, and its structure was characterized by various analytic methods. Immobilized lipase was applied for biodiesel production using acidified palm oil as the starting material, the conversion rate achieved as high as 98.5% in two steps. Furthermore, the immobilized lipase displayed satisfactory stability and reusability in biodiesel production. When the aforementioned reaction was carried out in a continuous flow packed bed system, the yield of biodiesel was 94.8% and space–time yield was 2.88 g/(mL∙h). The immobilized lipase CALB@resin-CAB showed high catalytic activity and stability, which has good potential for industrial application in the field of oil processing.
研究领域:
生物化学
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酶工程
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可再生能源
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PubMed
高影响因子
Catalysts
·
2026
·Vol. 16
·期 5
·377
·
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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高影响因子
Catalysis Science & Technology
·
2018
·Vol. 8
·期 18
·4718-4725
·
IF 5.365
Jiang-Wei Shen,Jia-Mei Qi,Xiao-Jian Zhang,Zhi-Qiang Liu,Yu-Guo Zheng
Candida antarctica lipase B (CALB) is a versatile and robust lipase with high activity and enantioselectivity in the resolution of alcohols and amines. Poor specific activity toward carboxylic esters that are branched or substituted in close proximity to the carboxyl group limits the application of CALB. In this study, a semi-rational engineering approach was successfully applied to tailor CALB for efficient synthesis of the moxifloxacin chiral intermediate through enzymatic resolution of cis-(±)-dimethyl 1-acetylpiperidine-2,3-dicarboxylate. The combinatorial active-site saturation test was performed on CALB and two key residue sites, 189 and 190, were identified in the determination of hydrolytic activity. The specific activity and catalytic efficiency (kcat/Km) of the best variant mut-I189K were increased by 286- and 193-fold, respectively, without impacting its enantioselectivity. With 0.1 g L−1 of purified mut-I189K loading, 1 M substrate was asymmetrically hydrolyzed with 49.9% conversion within 5 h, resulting in a space–time yield of 583.8 g L−1 d−1. These results demonstrated that mut-I189K is a competitive biocatalyst for large-scale preparation of the moxifloxacin chiral intermediate.
研究领域:
生物催化
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酶工程
·
药物化学
·
有机合成
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高影响因子
危险品化学品经营许可证（带存储）