INTERNATIONAL DAIRY JOURNAL
·
2026
·
IF 3.1
Weijia Bao,Jiaheng Li,Yingming Zhang,Yingxing Li,Ziyuan Chen,Menghui Sun,Anqi Chen,Xiaosan Wang,Yanfang Sun
Although low-fat dairy products are healthier, they usually have a poorer flavor compared to full-fat dairy products. In this study, lipase-catalyzed partial hydrolysis was employed to enhance flavor characteristics of anhydrous milk fat (AMF). Results showed that among the 24 evaluated lipases, Lipase A “Amano” 12 (A12), Newlase F3G (F3G), Lipozyme RM IM (RM IM), and Lipozyme Palatase 20000L (PAL) were the most effective lipases in enhancing dairy flavor. Nine key volatile organic compounds (VOCs) were identified by random forest (RF)-driven comprehensive approaches as the key flavor volatiles in the hydrolysates produced by the above four lipases, including short- and medium-chain fatty acids (SMCFAs), esters, and ketones. Results showed that RF models had excellent performance in evaluating the contributions of VOCs to sensory attributes based on their concentrations. The main novelty of this study lies in the expansion of lipase selection for enhancing AMF flavor.
✓ 已验证
产品关联
DOI
高影响因子
Scientific Reports
·
2026
·
IF 4.9
Yang Lutong,Wang Mengai,Fu Hongyan,Zhang Yun,Zhang Yu,Cui Mengni,Jiang Bowen,Chen Hong,Cui Yanwei
Cold-adapted lipases have attracted extensive research attention in enzymology owing to their unique low-temperature adaptability and catalytic properties, and investigations in this area continue to deepen. This study aimed to efficiently produce a cold-adapted lipase to overcome the limitations of conventional lipases under low-temperature conditions, such as low activity, high cost, and restricted applicability. Fungal strains were isolated from naturally cold environments, and subjected to multiple rounds of screening for lipase activity. A selected strain was further improved by UV-nitrosoguanidine (NTG) combined mutagenesis, followed by optimization of fermentation conditions. The lipasewas purified by ammonium sulfate precipitation, Q-HP anion exchange chromatography, and Superdex 75 gel filtration chromatography, and its enzymatic properties were characterized. A lipase-producing strain identified as Aspergillus niger was obtained through screening. Following combined mutagenesis, lipase activity by the mutant strain increased by 127.35%. Under the optimized fermentation conditions (temperature 21℃, pH 6.0, inoculation size 6.5 × 10 6 spores/mL), the lipase activity further increased to 57.26 U/mL. After purification, the cold-adapted lipase exhibited a specific activity of 113.52 U/mg. Its optimal reaction temperature was 30℃ and pH 7.0 and retained more than 60% of its maximum activity at 15℃. Its high relative activity at low temperatures, together with its low thermal stability, supported its classification as a cold-adapted lipase. This purification strategy can produce high-purity lipase at laboratory scale, which provides a pure enzyme preparation for biochemical studies. The cold-adapted lipase obtained in this study may have promising application prospects in fields such as food processing, low-temperature laundry, and environmental management.
✓ 已验证
DOI
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2024
·
IF 3.1
Zhang Yueting,Nie Hongmei,Zhang Fei,Jin Mengmeng,Wang Zhao,Zheng Jianyong
Aspergillus oryzae is an ideal cell factory for protein expression with powerful protein processing and secretion capabilities. The current study aimed to explore the homologous expression of A. oryzae lipase AOL (GenBank: KP975533) by constructing an auxotrophic A. oryzae △ pyrG △ nptB and subsequently characterizing the immobilization and catalytic properties of recombinant lipase. Initially, the pyrG gene knocked out in wild-type A. oryzae by homologous recombination, followed by the creation of a uridine/uracil auxotroph transformation. Through this system, the protease gene nptB was precisely knocked out, leading to a substantial decrease in extracellular (39.04%) and intracellular (90.07%) protease activity. The A. oryzae △ nptB △ pyrG strain was used as host for homologous expression of lipase AOL. After transformation of linearized lipase-expression cassette, the engineered A. oryzae AOL-8 was screened out with the lipase gene copy number of 14, exhibiting extracellular and intracellular lipase activities of 1.75 U/mL and 46.4 U/g, respectively. Subsequently, the production and immobilization of the recombinant lipase, via physical adsorption on macroporous resin XRZ04B, were achieved through submerged fermentation of the AOL-8 strain. The results of esterification catalytic properties of immobilized recombinant lipase indicated that the lipase exhibited optimal catalytic activity with lauric acid and methanol as substrates, a reaction temperature of 35 °C, and n-hexane as the preferred solvent medium; its highest conversion rate can reach at 72.3%.
研究领域:
微生物学
·
生物技术
·
酶学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LANGMUIR
·
2026
·
IF 4.4
Yudan Chai,Bin Wang,Jun Xu,Jian Zhang
Microbial lipases are versatile biocatalysts that are essential for hydrolyzing ester bonds in triglycerides. Among these enzymes, Pseudomonas cepacia lipase (PCL) possesses a canonical lid domain, whereas Candida antarctica lipase B (CALB) features a rigid lid-like structure. Understanding how these different structures catalyze mechanisms differently at the interface is crucial. Experimental assays demonstrated high activity for both enzymes in aqueous systems, but CALB exhibited lower activity than PCL in biphasic water–organic solvent systems. Molecular dynamics simulations were employed to investigate the conformational dynamics and interfacial interactions. The results reveal that the PCL undergoes polarity-induced conformational changes leading to channel shortening and bottleneck widening, thereby facilitating substrate access. In contrast, CALB retains a rigid architecture with fixed distances between the lid-like region and the catalytic triad, thereby limiting interfacial adaptability. PCL further employs a lid-mediated adsorption mechanism that enhances interfacial binding and exposes hydrophobic residues. Conversely, CALB relies on an active-site-oriented approach with a weaker interfacial affinity. Additionally, the aqueous environment promotes lid closure in lipase PCL by stabilizing hydrogen bond networks and inducing conformational shifts in key flexible regions. This study elucidates polarity-driven interfacial activation mechanisms distinguishing lid-containing and lid-like lipases, offering atomistic insights for rational lipase engineering in nonaqueous environments.
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2025
·Vol. 48
·期 4
·e70125
·
IF 2.8
Sikai Wang,Yue Li,Lei Wang,Lin Lv,Hongmei Liu,Guoqi Zhang,Yan Zhao
In this study, a novel ionic liquid/chitosan bifunctionalized magnetic multi-walled carbon nanotubes composite material (m-MWCNTs@CS@IL) was utilized for the first time to immobilize lipase. The immobilized lipase exhibited exceptional stability and reusability, as evidenced by its characteristic properties. A ligand fishing approach utilizing the immobilized lipase was developed to enable rapid screening of lipase inhibitors from Chrysanthemum . Three ligands were successfully screened from Chrysanthemum and then identified as cynaroside, quercitrin, and linarin by ultra-high performance liquid chromatography-tandem mass spectrometry. The IC 50 values of these three ligands were 76.77 ± 0.36, 83.01 ± 0.46, and 43.61 ± 0.77 µM, respectively. Furthermore, molecular docking analysis further confirmed the binding of three ligands to specific amino acid residues within the active site of lipase. This work presents a fast and efficient approach for screening lipase inhibitors from intricate natural sources, demonstrating promising prospects in discovering anti-obesity compounds.
研究领域:
生物化学
·
药物发现
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Wenxiang Hu,Jiangyu Zong,Jiajun Gong,Rong Xue,Yangbin Lv,Biao Liu,Wenting Dai,Yajun Wang,Chu Chu
Lipase immobilization plays a pivotal role in enhancing enzyme stability, reusability, and practical applicability in ligand fishing, a technique used to identify bioactive compounds from traditional Chinese medicine (TCM). However, conventional methods often lead to significant activity loss due to suffering from enzyme leakage, conformational distortion, and mass transfer resistance. In this study, we present a dynamic defect-engineering (DDE) strategy within zeolitic imidazolate framework-8 (ZIF-8) to overcome these limitations. The DDE process enables adaptive formation of transport channels driven by enzyme–framework interactions, facilitating the migration of lipase from surface adsorption to intracrystalline encapsulation. Brunauer–Emmett–Teller (BET) analysis confirmed successful encapsulation, showing a reduced specific surface area of 565.1 m 2 /g for lipase@ZIF-8, indicative of enzyme incorporation within the defect-engineered framework. This structural optimization significantly improved biocatalytic performance, achieving 82 % activity retention and 65 % immobilization yield, compared to 44 % activity retention in conventional in situ encapsulation. Utilizing the preserved catalytic activity, we established a lipase-based ligand fishing platform, which successfully identified 20 potential inhibitors (10 anthraquinones and 10 flavonoids) from Cassiae Semen and Citri Reticulatae Pericarpium using HPLC-Q-TOF-MS/MS analysis. Overall, this work demonstrates the effectiveness of the DDE strategy in enhancing lipase immobilization within ZIF-8, offering a robust and efficient platform for the high-throughput screening of lipase inhibitors from natural products and advancing the application of enzyme-based ligand fishing in TCM research.
研究领域:
酶工程
·
纳米材料
·
植物化学
·
药物发现
·
中医药研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2025
·
IF 9.8
Ying Li,Yu Zhang,Li Zhang,Jun Dang
The screening of enzyme inhibitors faces challenges such as low efficiency. In this study, a novel online HPLC-FLD/pancreatic lipase recognition system was developed for the first time. The optimal operating parameters were established as a flow rate of 1 mL/min, with excitation and emission wavelengths for pancreatic lipase set at 221 nm and 303 nm, respectively. The effectiveness of this system was validated using orlistat. Subsequently, this system was applied to Lycium ruthenicum , where two petunidin-based anthocyanins pancreatic lipase inhibitors were efficiently isolated using medium-pressure liquid chromatography. Molecular docking elucidated that compounds 1 and 2 interact with the serine residue of pancreatic lipase, exhibiting binding energies of −13.12 kcal/mol and −12.22 kcal/mol, respectively. Enzymatic assays showed that the half-maximal inhibitory concentrations of compounds 1 and 2 were 9.67 μM and 147.37 μM, respectively. These docking and inhibition results confirm the system for efficiently discovering pancreatic lipase inhibitors.
研究领域:
植物化学
·
分析化学
·
酶学
·
天然产物化学
·
药物发现
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MOLECULES
·
2022
·Vol. 27
·期 11
·3475
·
IF 4.927
Jinfang Xu,Pengkun Cao,Zhiyu Fan,Xujing Luo,Gangqiang Yang,Tingli Qu,Jianping Gao
As for ligand fishing, the current immobilization approaches have some potential drawbacks such as the small protein loading capacity and difficult recycle process. The core–shell metal–organic frameworks composite (Fe3O4[email protected]2), which exhibited both magnetic characteristics and large specific surface area, was herein fabricated and used as magnetic support for the covalent immobilization of porcine pancreatic lipase (PPL). The resultant composite Fe3O4[email protected]2@PPL manifested a high loading capacity (247.8 mg/g) and relative activity recovery (101.5%). In addition, PPL exhibited enhanced tolerance to temperature and pH after immobilization. Then, the composite Fe3O4[email protected]2@PPL was incubated with the extract ofScutellaria baicalensisto fish out the ligands. Eight lipase inhibitors were obtained and identified by UPLC-Q-TOF-MS/MS. The feasibility of the method was further confirmed through an in vitro inhibitory assay and molecular docking. The proposed ligand fishing technique based on Fe3O4[email protected]2@PPL provided a feasible, selective, and effective platform for discovering enzyme inhibitors from natural products.Keywords:ligand fishing;magnetic MOF;Scutellaria baicalensis;pancreatic lipase inhibitor
研究领域:
生物化学
·
药学
·
材料科学
✓ 已验证
DOI
PubMed
高影响因子
Food Bioscience
·
2026
·
IF 6.2
Hui Zhang,Mengjia Xu,Felix Wambua Muema,Jun Ding,Mingquan Guo
Chrysanthemum indicum L. has traditionally been utilized as one of the main ingredients in many functional foods or tea for lipid-lowing by advantages of abundant bioactive components and lower side effects. However, the specific compounds responsible for these beneficial lipid-lowering properties remain unrevealed. In this study, lipase-immobilized hollow fibers were first developed to screen out potential lipase ligands by integrating ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-QTOF-MS/MS) with a molecular networking approach, in which the incorporation of glutaraldehyde as crosslinker achieved a 2.24-fold increase in enzyme loading capacity. The immobilized lipase retained over 50% relative viability after 6 enzymatic cycles or 10 h of storage (4 °C). A total of 30 potential ligands were successfully screened out and identified. Among them, the inhibitory activities and the mechanism of action of three representative candidate ligands (Ethyl chlorogenate, Luteolin 7- O -glucuronide and Ethyl caffeate) were further investigated through in vitro enzyme activity assays and molecular docking analysis. Overall, the current study proposed a promising strategy for the rapid discovery and identification of bioactive compounds against obesity from complex natural extracts in natural foods.
✓ 已验证
DOI
高影响因子
ACS Sustainable Chemistry & Engineering
·
2024
·
IF 7.1
Lili Li,Xiaojing Wang,Yali Hu,Wang Sun,Yugao Ding,Nisha He,Guofu Zhou,Zhen Zhang
Enzymes are promising biocatalysts due to their high efficiency, mild conditions, high specificity, nontoxicity, and environmental friendliness. However, the enzyme suffers from poor stability and reusability. Enzyme immobilization is a commonly used technology to enhance its stability and reusability. In this study, the cellulose nanocrystal (CNC) was chosen as the enzyme immobilization carrier due to its sustainability, high specific surface area, biocompatibility, water dispersibility, and excellent Pickering emulsifying ability. Lipase was immobilized on a polydopamine-coated CNC (PC), and the obtained lipase-immobilized PC (LPC) displayed significantly enhanced stability and higher activity in harsh conditions compared to free lipase. LPC possessed partial wettability with both water and oil phases and a lower absolute value of the zeta potential than CNC, endowing it with a better Pickering emulsifying ability than CNC. LPC was fixed at the oil–water interface of the Pickering emulsion by ultrasonication, and the resulting Pickering emulsion droplets were employed as microreactors for ester hydrolysis and synthesis via Pickering interface biocatalysis. LPC displayed significantly enhanced catalysis activity at the oil–water interface of the Pickering emulsion compared to free lipase in a traditional oil–water biphasic system due to the increased contact area between the enzyme and the substrate, reduced diffusion distance of the substrate, and lipase interface activation effect. Moreover, LPC could be facilely recycled by centrifugation, and the recycled LPC still retains high catalysis activity. This study proposed a facile and sustainable method of enzyme immobilization to enhance its stability and reusability and achieved a significantly increased activity via Pickering interface biocatalysis.
研究领域:
生物技术
·
化学工程
·
材料科学
·
酶学
✓ 已验证
产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Yingao Zhai,Kunshan Wang,Zenghui Yu,Suzhen Zhou,Jinbo Fan
In this study, 3D-QSAR pharmacophore models were used to screen Chrysin (Chr), Genistein (Gen) and Naringenin (Nar) as inhibitors of pancreatic lipase (PL). The mechanisms of inhibition were explored using PL activity assays, enzyme kinetics, multispectral techniques, molecular docking, and molecular dynamics (MD) simulations, leading to the construction of a pharmacophore model that included two hydrogen bond acceptors and one hydrophobic group. Chr, Gen, and Nar competitively and reversibly inhibited PL with IC 50 values of 22.98 ± 1.45, 42.36 ± 0.35, and 48.66 ± 0.25 μmol·L −1 , respectively, and inhibition constants of (6.2995 ± 0.01) × 10 −5 , (6.5494 ± 0.02) × 10 −5 and (2.4361 ± 0.02) × 10 −5 mol·L −1 , respectively. Kinetic analysis, fluorescence quenching assays and molecular docking showed that thee compounds interacted with PL close to its active site (Ser153-His264-Asp177), affecting the microenvironment of tryptophan residues, and bound stably to PL via hydrogen bonds, hydrophobic interactions, and van der Waals forces. UV–visible spectra showed binding constants of 65.40 × 10 3 , 62.70 × 10 3 and 6.41 × 10 3 L/mol, respectively. MD simulations indicated that the structures of the complexes were compact, with binding free energy analysis ranking the PL inhibitory abilities of the compounds as Chr > Gen > Nar. In summary, the pharmacophore model was accurate. Chr, Gen, and Nar could inhibit PL activity. These findings provide a reference for the screening and development of new PL inhibitors.
研究领域:
药物化学
·
生物化学
·
计算生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
·
2016
·
IF 2.189
Chen Chen,Xue-yan Zhu,Qiao-ling Gao,Fei Fang,Li-wei Wang,Xiao-jun Huang
Functional polyphosphazene cyclomatrix microspheres (PMS) containing glucose and dodecyl moieties were utilized for physical adsorption of lipase. In this regard, Candia Rugosa lipase was immobilized onto the above microspheres through physical adsorption, which exhibited catalytic hydrolysis of glycerol triacetate as the model reaction. As observed under transmission electron microscope (TEM) and scanning electron microscope (SEM), the size of all the microspheres ranged from 400 nm to 1 μm with a mean diameter of 680 nm. Zeta potential analysis demonstrated that the microspheres were negatively charged. The influences of buffer pH, substrate concentration and temperature were also studied along with reusability. The obtained results demonstrated that the immobilized lipase on deprotected glycosylated PMS (D-GPMS) showed the best performance and possessed the highest enzymatic loading activity around 1.0884 mmol L −1 min −1 g −1 , and retained nearly 50% of its initial activity even after 10 cycles of application. Moreover, the lipase-microsphere complex displayed high pH adaptability from 7 to 9 and temperature stability above 40 °C. From the viewpoint of biocompatible PMS support, the GPMS-lipase conjugate have potential applications in field such as food, medicine and environment.
研究领域:
生物材料
·
酶工程
·
纳米医学
·
分子生物学
·
环境生物技术
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）