CHEMICAL ENGINEERING JOURNAL
·
2023
·
IF 15.1
Hao Zhou,Yongheng Shi,Lingmei Dai,Dehua Liu,Wei Du
Pyrolyzing ZIF-8 into N-doped carbon is now inspiring new potential of ZIF-8 for extensive applications. However, the exploration of such ZIF-8-derived carbon for biotechnological applications has been rarely investigated, especially in this context that imidazole derivatives are of rich functionality in life science. Herein, we reported a facile temperature-dependent pyrolysis method to achieve the effective N-doping in ZIF-8-derived macroporous carbon, which was systematically investigated for the immobilization of lipase from Thermomyces lanuginosus (TLL) in biodiesel production. MNC-600 (macroporous N-doped carbon derived at 600 °C) exhibited the best immobilization performance, with the highest specific activity of 190.6 U mg −1 , a 36.9% increase than free TLL. Moreover, MNC-600@TLL showed superior effectiveness and robustness in biodiesel production. A much faster rate of conversion and the highest biodiesel yield of 87.4% were observed. After eight successive batches of recycling, MNC-600@TLL retained 93.4% of its initial activity. XPS analysis revealed the evolution of N species during the pyrolysis of ZIF-8. It was proposed that the abundant doping of pyrrolic N species played a major role in the surprising performance of MNC-600, mainly through hydrogen bonding interactions, which simultaneously realized the activation of reactants as well as the in-situ hydrophobization of biocatalyst. This report is anticipated to pave a way for the exploration of ZIF-8-derived N-doped carbon for extensive biotechnological applications.
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FOOD CHEMISTRY
·
2022
·
IF 9.231
Xuan Liu,Weifei Wang,Zexin Zhao,Long Xu,Bo Yang,Dongming Lan,Yonghua Wang
Enzymatic degumming is an essential refining process to improve oil quality. In this study, a monoacylglycerol lipase GMGL was derived from marine Geobacillus sp., and was found that not only took monoacylglycerol (MAG) as substrate, but also had activity toward lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE) and glycerolphosphatidylcholine (GPC). Binding free energy showed LPC and LPE could bind with enzyme stably as MAG. It presented great potential in the field of enzymatic degumming. The phosphorus content in crude soybean oil decreased from 680.50 to 2.01 mg/kg and the yield of oil reached to 98.80 % after treating with phospholipase A1 (Lecitase Ultra) combined with lipase GMGL. An ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was developed to identify 21 differential phospholipids between crude soybean oil and enzymatic treatment. This work might shed some light on understanding the catalytic mechanism of monoacylglycerol lipase and provide an effective strategy for enzymatic degumming.
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Evidence-based Complementary and Alternative Medicine
·
2022
·
IF 2.65
Hao Zhang,Feng-Qing Yang,Jian-Li Gao
With the increase of obesity incidence, the development of antiobesity drugs has aroused extensive interest. In this study, a simple and portable personal glucose meter (PGM) method based on the lipase-mediated reaction combined with molecular docking was developed for the screening of lipase inhibitors. Lipase can catalyse the hydrolysis of 4-acetamidophenyl acetate to form acetaminophen, which can directly trigger the reduction of K 3 [Fe(CN) 6 ] to K 4 [Fe(CN) 6 ] in the glucose test strips and generate an electrical signal that can be detected by the PGM. When lipase inhibitors exist, the yield of acetaminophen will be reduced and results in a corresponding decrease of the PGM signal. Therefore, the activity of lipase can be measured by the PGM. After optimization of the experimental conditions, the inhibitory activity of fourteen small-molecule compounds and fifteen natural product extracts on lipase were evaluated by the developed PGM method. The results indicate that tannic acid, (-)-epigallocatechin gallate, (-)-epigallocatechin, (-)-epicatechin gallate, and epicatechin have good inhibitory effect on lipase (% of inhibition higher than 40.0%). Besides, the natural product extracts of Galla Chinensis, lemon, and Rhei Radix et Rhizoma have a good inhibitory effect on lipase with % of inhibition of (97.5 ± 0.6)%, (88.1 ± 0.7)%, and (79.1 ± 1.6)%, respectively. Finally, the binding sites and modes of six small-molecule compounds on lipase were investigated by the molecular docking study. The results show that the developed PGM method is an effective approach for the discovery of potential lipase inhibitors.
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Environmental Technology & Innovation
·
2022
·
IF 7.758
Jianqiang Sun,Haofeng Zhu,Xindong Yang,Yan Zheng,Tianchi Sun,Honglei Xu,Jiaqi Meng,Anping Zhang
Phthalate esters (PAEs) are ubiquitous in agricultural soil and crops because of the widespread application of plastic films containing PAEs. The enzymes play critical roles in the elimination of pollutants from environmental matrices. In an attempt to evaluate the effect of enzymes on the degradation of various PAEs, this study investigated the enzyme-catalyzed reaction kinetics of seven selected PAE congeners in water and soil. Carboxylesterase and lipase catalyzed the transformation of PAEs to monoalkyl phthalate esters and phthalic acid with the half-life ranges of 4.78–126.03 h. PAEs with long-alkyl-chain were more recalcitrant to carboxylesterase and lipase-catalyzed degradation, and PAEs with short alkyl chains were relatively easier to be degraded by the two enzymes. The results indicate that the alkyl chain length had a profound effect on the enzyme-catalyzed degradation of PAEs. The reaction rates and half-lives were significantly correlated with the log Kow value ( p < 0.01), which was dominated by the molecular structure. The results obtained in this work are expected to well guide the application of carboxylesterase and lipase in the remediation of waterbody contaminated by various PAEs. With respect to soil, they can be well used in the remediation, exposure, and risk assessment of paddy soil. However, the application of the results in the remediation of other soils might lead to the risk of a low removal ratio.
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BIOPROCESS AND BIOSYSTEMS ENGINEERING
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2022
·Vol. 45
·期 4
·749-759
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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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Frontiers in Bioengineering and Biotechnology
·
2022
·
IF 6.064
Xue Yu,Zhang Xiao-Guang,Lu Ze-Ping,Xu Chao,Xu Hua-Jin,Hu Yi
Various betaine ionic liquids composed of different chain lengths and different anions were designed and synthesized to modify Candida antarctica lipase B (CALB). The results showed that the catalytic activity of all modified lipases improved under different temperature and pH conditions, while also exhibiting enhanced thermostability and tolerance to organic solvents. With an increase in ionic liquid chain length, the modification effect was greater. Overall, CALB modified by [BetaineC16][H2PO4] performed best, with the modified CALB enzyme activity increased 3-fold, thermal stability increased 1.5-fold when stored at 70°C for 30 min, with tolerance increased 2.9-fold in 50% DMSO and 2.3-fold in 30% mercaptoethanol. Fluorescence and circular dichroism (CD) spectroscopic analysis showed that the introduction of an ionic liquid caused changes in the microenvironment surrounding some fluorescent groups and the secondary structure of the CALB enzyme protein. In order to establish the enzyme activity and stability change mechanisms of the modified CALB, the structures of CALB modified with [BetaineC4][Cl] and [BetaineC16][Cl] were constructed, while the reaction mechanisms were studied by molecular dynamics simulations. Results showed that the root mean square deviation (RMSD) and total energy of modified CALB were less than those of native CALB, indicating that modified CALB has a more stable structure. Root mean square fluctuation (RMSF) calculations showed that the rigidity of modified CALB was enhanced. Solvent accessibility area (SASA) calculations exhibited that both the hydrophilicity and hydrophobicity of the modified enzyme-proteins were improved. The increase in radial distribution function (RDF) of water molecules confirmed that the number of water molecules around the active sites also increased. Therefore, modified CALB has enhanced structural stability and higher hydrolytic activity.
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ACS Biomaterials Science & Engineering
·
2023
·
IF 5.8
Chang Liu,Qing Yang,Hong Lin,Limin Cao,Kaiqiang Wang,Jianxin Sui
Nowadays, obesity severely impacts human health and is the fifth leading risk factor that leads to death globally. Pancreatic lipase (PL) inhibitors have attracted extensive attention owing to their role in effective prevention and treatment of obesity. Here, a shark-derived single-domain antibody fusion protein was used to inhibit PL for the first time. After biopanning, the heterologous expression system pET28a-SUMO-D2 was constructed using the method of double restriction enzyme digestion and T4 ligase to achieve the soluble expression of the PL-specific antibody gene D2. According to the calculation of protein concentration, the final expression of fusion protein PL-D2S was 1.183 mg per liter of Luria Bertani broth. The binding ability of the soluble fusion protein PL-D2S to PL was identified. Enzyme-linked immunosorbent assay results showed that the fusion protein PL-D2S exhibited a strong binding affinity to PL. The experimental results of PL inhibition of PL-D2S in vitro showed that the fusion protein could significantly inhibit the activity of PL, with an IC50 of 404 μg/mL. Our study shows that the fusion protein PL-D2S is a potential PL inhibitor to prevent and treat obesity.
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FOOD CHEMISTRY
·
2023
·
IF 8.8
Fangfang Huang,Qingfei Dai,Kewei Zheng,Qingbao Ma,Yu Liu,Shuoqi Jiang,Wei Jiang,Xiaojun Yan
The effective modulation of pancreatic lipase and cholesterol esterase activities proves critical in maintaining circulatory triglycerides and cholesterol levels within physiological boundaries. In this study, peptides derived from KPHs-AL, produced through the enzymatic hydrolysis of skipjack tuna dark muscle using alkaline protease, have a specific inhibitory effect on pancreatic lipase and cholesterol esterase. It is hypothesized that these peptides target and modulate the activities of enzymes by inducing conformational changes within their binding pockets, potentially impacting the catalytic functions of both pancreatic lipase and cholesterol esterase. Results revealed these peptides including AINDPFIDL, FLGM, GLLF and WGPL, were found to nestle into the binding site groove of pancreatic lipase and cholesterol esterase. Among these, GLLF stood out, demonstrating potent inhibition with IC 50 values of 0.1891 mg/mL and 0.2534 mg/mL for pancreatic lipase and cholesterol esterase, respectively. The kinetics studies suggested that GLLF competed effectively with substrates for the enzyme active sites. Spectroscopic analyses, including ultraviolet–visible, fluorescence quenching, and circular dichroism, indicated that GLLF binding induced conformational changes within the enzymes, likely through hydrogen bond formation and hydrophobic interactions, thereby increasing structural flexibility. Molecular docking and molecular dynamics simulations supported these findings, showing GLLF's stable interaction with vital active site residues. These findings position GLLF as a potent inhibitor of key digestive enzymes, offering insights into its role in regulating lipid metabolism and highlighting its potential as functional ingredient.
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JOURNAL OF POLYMER RESEARCH
·
2023
·Vol. 30
·期 11
·1-18
·
IF 2.8
Li Xin,Zhan Shiping
Poly (L-lactic acid) (PLLA) is a bio-based biodegradable polymer with minimal immunogenicity, non-toxicity and excellent mechanical properties and has widely used in medical, pharmaceutical, food packaging and other fields. However, linear PLLA has the characteristics of high melting point, strong crystallinity, weak flowability and poor thermal stability, which cannot meet the requirements of various industrial processes and novel pharmaceutical formulations. With its low viscosity, controlled degradation cycle, and excellent melt flowability, star-shaped PLLA can be used in the fields of biocomposites, biomedical devices, drug delivery, and intelligent packaging. Presently, metal catalysts and organic solvents are frequently used in PLLA synthesis industries, and both residues continue to pose a safety risks. In existing studies, the performance of bioenzymes as optimal green catalysts in organic solvents is not satisfactory, and they are predominantly used to synthesise oligomers with low yields and lengthy reaction times. In this study, a four-armed star-shaped poly (L-lactic acid) (4s-PLLA) with a porous structure favourable for drug loading was synthesised using supercritical carbon dioxide (Sc-CO 2 ) as a solvent, lipase as a catalyst, and erythritol as a nucleus. Using Sc-CO 2 as a solvent the reaction activity and stability of the enzyme were increased so that the reaction yield could reach 83% and the reaction time was only 24 h, which provided a possibility for the practical application of the enzymes catalyze green process. Both the random copolymerization product four-armed star-shaped poly(L-lactic-glycolic acid) (4s-PLGA) and the block copolymerization product four-armed star-shaped poly (L-lactic-glycolic acid) (4s-PLLA-PGA) were subsequently synthesized by copolymerization modification, and their porous structures retained and hydrophilicity were enhanced. The response surface analyses were performed about the reaction time, reaction temperature and yield of synthetic reactions by using Design-Expert 12 software and the prediction equations were generated.
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FOOD CHEMISTRY
·
2023
·
IF 8.8
Lili Yin,Kunpeng Gao,Xiangzhao Mao,Yang Hu
Lipase B from Candida antarctica (CALB) plays a prominent role as a biocatalyst in several industries, especially for biphasic conversion of functional lipids. Herein, an amphiphilic Janus halloysite nanosheet (JHNS) was fabricated and employed simultaneously as a solid surfactant for stabilizing Pickering emulsion and as a carrier for immobilizing CALB, with the aim to realize highly efficient biphasic bioconversion. The obtained JHNS could stabilize Pickering emulsion for at least 7 days. Immobilization of CALB on JHNS improved the substrate affinity, catalytic efficiency, thermal stability, and alkaline tolerance of the enzyme. Moreover, JHNS-based immobilized CALB was exploited as a biocatalytic platform for the conversion of retinyl acetate, with almost twice increase in conversion efficiency. Taken together, the JHNS-based immobilized CALB paves the way for the design of efficient biphasic conversion system for the production of added-value lipids.
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LWT-FOOD SCIENCE AND TECHNOLOGY
·
2021
·
IF 4.952
Pengkai Wang,Peng Fei,Chunxia Zhou,Pengzhi Hong
This study investigated the acylation of pectin with p- coumaric and caffeic acids through an enzymatic method to enhance its physical and chemical properties. Ultraviolet–visible, Fourier transform infrared and 1 H nuclear magnetic resonance spectral analyses indicated the successful grafting of p- coumaric and caffeic acids onto the carboxyl group of pectin molecule. The 1,1-diphenyl-2-picryl hydrazine (DPPH) clearance and inhibition ratio in the β-carotene bleaching assay of native pectin (Na–Pe) were 0.13% and 42.60%, respectively, while their corresponding values increased to 1.04% and 55.59 for the pectin acylated with p -coumaric acid (Co–Pe) and 87.52% and 94.05% for the pectin acylated with caffeic acid (Ca–Pe). Meanwhile, the inhibition of Escherichia coli and Staphylococcus aureus was increased from 2.56% to 6.59% (Na–Pe) to 17.28% and 25.79% (Co–Pe) and 23.59% and 36.94% (Ca–Pe), respectively. Furthermore, Co-Pe or Ca-Pe coating was shown to significantly enhance the shelf life of small yellow croaker ( Larimichthys polyactis ) fillets.
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CHINESE JOURNAL OF CHEMICAL ENGINEERING
·
2021
·
IF 3.171
Chunyu Zhang,Yan Sun,Xiaoyan Dong
Enzyme-polymer conjugates are complex molecules with great practical significance. This work was designed to develop a novel enzyme-polymer conjugate by covalently coupling a zwitterionic polymer with side dimethyl chains (pID) to Candida rugosa lipase (CRL) via the reaction between the anhydrides of polymer chains with the amino groups of the enzyme. The resulting two CRL-pID conjugates with different pID grafting densities were investigated in term of the catalytic activity, stability and structural changes. In comparison with native CRL, both the CRL conjugates displayed 2.2 times higher activity than the native enzyme, and showed an increase in the maximum reaction rate ( V max ) and a decrease in the Michaelis constant ( K m ), thus resulting in about three-fold increases in the catalytic efficiency ( k cat / K m ). These are mainly attributed to the activation of lipase by the hydrophobic alky side chains. Moreover, the thermostability and pH tolerance of the lipase conjugates were significantly enhanced due to the stabilizing effect of the zwitterion moieties. For instance, a five-fold increase of the enzyme half-life at 50 °C for the high-pID conjugated CRL was observed. Spectroscopic studies reveal that the pID conjugation protected the enzyme in the changes in its microenvironment and conformation, well correlating with enhanced activity and stability of lipase conjugates. The findings indicate that enzyme conjugation to the zwitterionic polymer is promising for improving enzyme performance and deserves further development.
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