Foods
·
2023
·Vol. 12
·期 22
·4098
·
IF 5.2
Linlin Xu,Fei Pan,Yingnan Li,Huiqian Liu,Chengtao Wang
Lipase has great application potential in hydrolyzing residual yolk lipid in egg white liquid to restore its functional properties. In this study, a lipase gene fromBacillus subtiliswas expressed inE. coliBL21 (DE3) and named Lip-IM. Results showed that although Lip-IM has stronger specificity for medium- and short-chain substrates than long-chain substrates (C16, C18), due to its excellent enzyme activity, it also has strong hydrolysis activity for long-chain substrates and maintained over 80% activity at 4–20 °C, but significantly reduced when the temperature exceeds 40 °C. The addition of 0.5% Lip-IM enhanced foaming ability by 26% (from 475 to 501%) and reduced liquid precipitation rate by 9% (from 57 to 48%). Furthermore, molecular dynamics (MD) simulations were run to investigate the conformational stability of Lip-IM at different temperatures. Results showed that Lip-IM maintained a stable conformation within the temperature range of 277–303 K. Fluctuations in the flexible area and backbone movement of proteins were identified as the main reasons for its poor thermal stability.Keywords:lipase;foaming property;molecular dynamics simulation;thermal stability;dynamic cross-correlation analysis
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Biotechnology for Biofuels
·
2021
·Vol. 14
·期 1
·1-12
·
IF 6.04
Zhou Zi-Wen,Cai Chun-Xian,Xing Xiu,Li Jun,Hu Zu-E.,Xie Zong-Bo,Wang Na,Yu Xiao-Qi
Background Production of biodiesel from renewable sources such as inedible vegetable oils by enzymatic catalysis has been a hotspot but remains a challenge on the efficient use of an enzyme. COFs (Covalent Organic Frameworks) with large surface area and porosity can be applied as ideal support to avoid aggregation of lipase and methanol. However, the naturally low density limits its application. In this work, we reported a facile synthesis of core–shell magnetic COF composite (Fe 3 O 4 @COF-OMe) to immobilize RML ( Rhizomucor miehei lipase), to achieve its utilization in biodiesel production. Result This strategy gives extrinsic magnetic property, and the magnetic COFs is much heavier and could disperse in water medium well, facilitating the attachment with the enzyme. The resultant biocomposite exhibited an excellent capacity of RML due to its high surface area and fast response to the external magnetic field, as well as good chemical stability. The core–shell magnetic COF-OMe structure not only achieved highly efficient immobilization and recovery processes but also maintained the activity of lipase to a great extent. RML@Fe 3 O 4 @COF-OMe performed well in practical applications, while free lipase did not. The biocomposite successfully achieved the production of biodiesel from Jatropha curcas Oil with a yield of about 70% in the optimized conditions. Conclusion Magnetic COFs (Fe 3 O 4 @COF-OMe) for RML immobilization greatly improved catalytic performance in template reaction and biodiesel preparation. The magneticity makes it easily recovered and separated from the system. This first successful attempt of COFs-based immobilized enzyme broadened the prospect of biodiesel production by COFs with some inspiration.
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BIOCHEMICAL ENGINEERING JOURNAL
·
2019
·
IF 3.371
Chunyu Zhang,Yang Liu,Yan Sun
We recently reported that Candida rugosa lipase (CRL) bonded on zwitterionic polymer-grafted silica nanoparticles (SNPs-pOD-CRL) showed remarkably enhanced catalytic activity and substrate affinity. The polymer, pOD, with long cetane side chains, was a polymer derivative of poly(maleic anhydride- alt -1-octadecene) and N,N -dimethylethylenediamine. Here, a new zwitterionic polymer with shorter alkyl side chains was synthesized by reaction between poly(isobutylene- alt -maleic anhydride) and N,N -dimethylethylenediamine, denoted as pID, and grafted onto SNPs to develop a new zwitterionic support, SNPs-pID. CRL was coupled onto SNPs-pID, and SNPs-pID-CRL was extensively characterized and compared with SNPs-pOD-CRL. It was found that SNPs-pID-CRL presented higher enzyme-substrate affinity and catalytic activity than free lipase because of lipase activation by the hydrophobic alkyl chains of the polymer. Comparison with SNPs-pOD-CRL demonstrated that decreased hydrophobicity of the side chains was unfavorable for lipase activation, resulting in lower activity of lipase coupled on pID than on pOD. However, SNPs-pID-CRL exhibited significant increased thermal stability than SNPs-pOD-CRL because higher hydrophilicity of pID favored protein stabilization. These results indicate that a zwitterionic polymer with long alkyl side chains favor lipase activation rather than enzyme stability, while that with short alkyl side chains are beneficial to thermostability. Together, pID-functionalized support is promising for immobilized lipase of long-term applications. The work has thus provided new insights into the fabrication of thermostable immobilized lipase.
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ANALYTICA CHIMICA ACTA
·
2019
·
IF 5.256
Xinlin Chen,Shang Xue,Yaolan Lin,Jianguang Luo,Lingyi Kong
In terms of ligand fishing, the amount and the relative activity recovery of enzymes immobilized on magnetic particles and nanoparticles are not preeminent. Therefore, the metal-organic framework (MOF) UiO-66-NH 2 was synthesized to immobilize the porcine pancreatic lipase (PPL) via precipitation-cross-linking, and the resulting novel biological matrices named PPL@MOF manifested high PPL loading capacity (98.31 mg/g) and relative activity recovery (104.4%). Moreover, the novel enzyme-MOF composite was applied to screen lipase inhibitors from Prunella vulgaris L. to enrich and improve the techniques of ligand fishing. As a result, 13 lipase ligands were obtained, and 12 compounds were determined by HPLC-Q-TOF-MS/MS. All of these ligands were further confirmed to be potential inhibitors through the verification of the activity assay and molecular docking . The proposed approach based on PPL@MOF was superior in terms of abundant protein loading capacity, high enzyme catalytic activity and easy preparation process. Taken together, our newly developed method provided a new platform for efficient discovering bioactive molecules from natural herbs.
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THERMOCHIMICA ACTA
·
2018
·
IF 2.189
Yan Zhang,Xing-An Luo,Li-Juan Zhu,Shen-Zhi Wang,Ming-Qiang Jia,Zhong-Xiu Chen
Pancreatic lipase is an interfacial enzyme. The kinetic parameters of pancreatic lipase derived from heat release in crowded surroundings have not been reported. In this work, the effect of crowded medium on the catalytic activity of porcine pancreatic lipase (PPL) during lipid digestion was investigated by Isothermal Titration Calorimetry (ITC). The results showed that dietary polysaccharide Ficoll400 tended to increase the catalytic efficiency of PPL for both glyceryl trioleate and glyceryl trioctanoate substrates, whereas Dextran20 decreased the activity of the enzyme. The change in the fluorescence intensity and secondary structural content of PPL parallels the change of k cat / K m , of PPL, which was further confirmed in PEG2000. The findings of this research improve understanding of macromolecular crowding effects on interfacial catalysis by lipases. Moreover, it provides a useful method for research on enzyme kinetics in crowed medium by ITC .
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Green Chemistry Letters and Reviews
·
2024
·
IF 5.8
Hong Zhang,Wenbin Liu,Lei Wang,Xin Zhang,Bo Wang,Xu Zeng,Bin Zhang
The immobilization of enzymes is an important strategy to improve their stability and reusability. However, it often comes with a trade off of reduced biological activity and catalytic efficiency. In this study, a composite hydrogel based on corn stalk carboxymethyl cellulose, acrylic acid, and mesoporous silica SBA-15 was developed as a support for the immobilization of lipase from Pseudomonas S.P. (PSL). The surface morphology and functional properties of the composite hydrogels were investigated by scanning electron microscopy, Fourier transform infrared, X-ray diffraction and simultaneous thermal analyzer, respectively. The composite hydrogel materials had a large number of mesopores and macropores were present in the wall of the hydrogel sheet. The hydrogel had a network structure that reduced the diffusion resistance of the substrate and improved the accessibility of the enzyme. The immobilized PSL exhibited high enantioselectivity (E value: 88.2) and enzyme activity (316.8 µmol/g/min) in the preparation of (S)-ibuprofen by enzymatic hydrolytic resolution of racemic ibuprofen ethyl ester. This prepared novel composite hydrogel is a promising supporting matrix for the immobilization of enzymes.
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MOLECULES
·
2017
·Vol. 22
·期 12
·2157
·
IF 2.861
Weiwei Zhang,Huixia Yang,Wanyi Liu,Na Wang,Xiaoqi Yu
Lipases are the most widely employed enzymes in commercial industries. The catalytic mechanism of most lipases involves a step called “interfacial activation”. As interfacial activation can lead to a significant increase in catalytic activity, it is of profound importance in developing lipase immobilization methods. To obtain a potential biocatalyst for industrial biodiesel production, an effective strategy for enhancement of catalytic activity and stability of immobilized lipase was developed. This was performed through the combination of interfacial activation with hybrid magnetic cross-linked lipase aggregates. This biocatalyst was investigated for the immobilization of lipase fromRhizomucor miehei(RML). Under the optimal conditions, the activity recovery of the surfactant-activated magnetic RML cross-linked enzyme aggregates (CLEAs) was as high as 2058%, with a 20-fold improvement over the free RML. Moreover, the immobilized RML showed excellent catalytic performance for the biodiesel reaction at a yield of 93%, and more importantly, could be easily separated from the reaction mixture by simple magnetic decantation, and retained more than 84% of its initial activities after five instances of reuse. This study provides a new and versatile approach for designing and fabricating immobilized lipase with high activation and stability.Keywords:lipase;interfacial activation;surfactant;magnetic nanoparticles;cross-linked enzyme aggregates;enzyme immobilization;biodiesel
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FOOD CHEMISTRY
·
2017
·
IF 4.529
Mingming Zheng,Shi Wang,Xia Xiang,Jie Shi,Juan Huang,Qianchun Deng,Fenghong Huang,Junyong Xiao
In this study, Candida lipolytica lipase ( CLL ) was immobilized on magnetic multi-walled carbon nanotubes (mMWCNTs) via hydrophobic and cation-exchange interaction. The resultant immobilized CLL showed much better thermal stability, biocatalyst activity and easier recycling than the free form. A method for efficient enzymatic acidolysis of tripalmitin (PPP) with oleic acid (OA), to produce OPO-rich TAGs, was developed, using the immobilized CLL as the biocatalyst. Under optimized conditions (2% water, 20 mg/ml enzyme, 1:6 PPP/OA, 50 °C, 2 h), the content of OPO in the final product reached 46.5%. CLL @mMWCNTs had a better activity and manipulative stability than commercial lipases. More importantly, the feasibility of CLL @mMWCNTs was also validated in the practical production of OPO-rich TAGs, using lard and restructured palm oil as the raw material. These results suggest that CLL @mMWCNTs is a promising biocatalyst for the OPO-rich TAGs production and will be helpful for the infant formula industry.
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JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY
·
2013
·Vol. 91
·期 1
·55-62
·
IF 1.592
Qin Xiao-Li,Huang Hui-Hua,Lan Dong-Ming,Wang Yong-Hua,Yang Bo
Typoselectivity of crude CBD-T1 lipase ( Geobacillus sp. T1 lipase fused with a cellulose binding domain) was investigated. Multi-competitive reaction mixtures including a set of n -chain fatty acids (C8:0, C10:0, C12:0, C14:0, C18:1 n-9, C18:2 n-6 and C18:3 n-3) and tripalmitin-enriched triacylglycerols were studied in hexane. The crude CBD-T1 lipase discriminated strongly against C18:1 n-9 [competitive factor ( α ) = 0.23] and showed the highest preference for C8:0 ( α = 1). Utilizing the catalytic properties of crude CBD-T1 lipase, acidolysis of soybean oil with C8:0 was selected as a model reaction to investigate the ability of the lipase to produce MLM-type (medium-long-medium) structured lipids. Several reaction parameters (added water amount, reaction temperature, substrate molar ratio and reaction time) examined for incorporating C8:0 into soybean oil, the optimum conditions were: 1:3 (soybean oil/C8:0) of molar ratio, 3 mL of hexane, 50 °C of temperature, 48 h of reaction time, 20 % of crude CBD-T1 lipase (w/w total substrates), and 7.5 % of water (w/w enzyme). Under these conditions, the incorporation of C8:0 was 29.6 mol%. The results suggest that crude CBD-T1 lipase, which showed different fatty acid specificity profiles, is a potential biocatalyst for the modification of fats and oils.
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EUROPEAN JOURNAL OF ORGANIC CHEMISTRY
·
2014
·Vol. 2014
·期 14
·2917-2923
·
IF 3.154
Bo Xia,Guilin Cheng,Xianfu Lin,Qi Wu
Graphical A dynamic double kinetic resolution (DDKR) strategy under the co-catalysis of Raney nickel and Candida antarctica lipase B (CAL-B) was established for the one-pot simultaneous resolution of primary amines and secondary alcohols. Herein, we have established a dynamic double kinetic resolution (DDKR) strategy under the co-catalysis of Raney nickel and Candida antarctica lipase B (CAL-B) for the one-pot simultaneous resolution of primary amines and secondary alcohols (or esters). The DDKR strategy was successfully applied to the resolution of a series of racemic amines and secondary alcohols (or esters) as well as mexiletine, an important antiarrhythmic agent. The catalysts could be recycled and reused several times with the same high activity. Scale-up experiments were also successful. As a more atom-economical and efficient process than traditional simple kinetic resolutions, the DDKR strategy can be widely used to prepare optically pure amines and alcohols.
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FOOD CHEMISTRY
·
2021
·
IF 7.514
Junxin Zhao,Maomao Ma,Xianghui Yan,Dongman Wan,Zheling Zeng,Ping Yu,Deming Gong
The lipase from Bacillus licheniformis NCU CS-5 was immobilized onto β -cyclodextrin (CD) grafted and aminopropyl-functionalized chitosan-coated Fe 3 O 4 magnetic nanocomposites (Fe 3 O 4 -CTS-APTES-GA- β -CD). Fourier transform infrared spectroscopy, thermogravimetry analysis, X-ray diffraction, scanning electron microscopy and transmission electron microscopy showed that not only the functionalized magnetic nanoparticles were synthesized but also the immobilized lipase was successfully produced. The immobilized lipase exhibited higher optimal pH value (10.5) and temperature (60℃) than the free lipase. The pH and thermal stabilities of the immobilized lipase were improved significantly compared to the free lipase. The immobilized lipase remained more than 80% of the relative activity at temperature of 60 ℃ and pH 12.0. The immobilized lipase also remained over 80% of its relative activity after 28 days of storage and 15 cycles of application. The application of the immobilized lipase in esterification of isoamyl acetate and pentyl valerate showed that maximum esterification efficiency was achieved in n -hexane having 68.0% and 89.2% respectively. Therefore, these results indicated that the Fe 3 O 4 -CTS-APTES-GA- β -CD nanoparticles are novel carriers for immobilizing enzyme, and the immobilized lipase can be used as an innovative green approach to the synthesis of fruity flavor esters in food industry.
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BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2025
·
IF 3.6
Yue Guoliang,Cai Zhenling,Meng Chun,Mao Yuhong
Efficient immobilized lipase systems are essential for industrial lipid modification, yet the combined effects of enzyme loading, support properties, and post-immobilization stabilization on catalytic performance remain underexplored. In this study, Thermomyces lanuginosus lipase (TLL), Candida antarctica lipase B (CalB), and their mixture (T + C) were respectively immobilized on two structurally distinct commercial supports (A & B)—differing in hydrophobicity, surface area, and pore size—at varied loading densities. Adsorption followed monolayer kinetics, with support B enabling higher enzyme up to 44.1 mg/g wet for T + C. Co-immobilized T + C systems exhibited enhanced catalytic activity compared to individual lipases, particularly in esterification and transesterification, benefiting from the complementary substrate specificities of TLL and CalB. Chemical crosslinking with chitosan and glutaraldehyde significantly reduced lipase desorption and improved thermal stability. The optimized system Cr-B-T + C-M (crosslinked support B-TLL + CalB with medium enzyme loading) efficiently catalyzed fish oil transesterification at 60 °C for 18 h, increasing the EPA + DHA content from 30.5% to 54.8%. All immobilized lipases retained over 80% of their initial catalytic activity after three reuses (60 °C and 18 h). These results highlight the synergistic benefits of co-immobilization and crosslinking on tailored supports, offering a versatile strategy for developing stable and efficient lipase biocatalysts for high-value lipid processing.
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