JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2026
·
IF 6.7
Yuchen Wang,Lu Wang,Anjun Li,Jiangjing Gao,Shuyang Hu,Yan Xu,Xiao-Wei Yu
The thermostability of lipases represents a key advantage for expanding their application potential in the food industry. In this study, a thermotolerant lipase, rTaLip, was identified from Baiju Daqu and successfully expressed in Komagataella phaffii. After determining its enzymatic properties, we developed two thermostable mutants through rational engineering (V219C/D222C/S73F/V141T and V219C/D222C/S73F/V141D). Compared to the wild-type, the mutants showed 2.2-fold and 4.1-fold longer half-lives at 65 °C, and their catalytic efficiency increased to 455% and 119%, respectively. Molecular dynamics simulations elucidated the structural basis underlying the increased thermal stability and catalytic activity. In application tests, the mutant V219C/D222C/S73F/V141T improved palm oil hydrolysis efficiency from 63.01% to 88.77%. This study demonstrates that combining the mining of thermostable lipases from Baijiu Daqu with rational design-based engineering constitutes a valuable strategy for improving the industrial applicability of lipases in harsh processing environments.
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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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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Hang Ning,Wen-long Liu,Qing-yun Li,You-yan Liu,Shi-ting Huang,Hai-bo Liu,Ai-xing Tang
Synthetic aromatic esters, widely employed in agriculture, food, and chemical industries, have become emerging environmental pollutants due to their strong hydrophobicity and poor bioavailability. This study attempted to address this issue by extracellularly expressing the promiscuous aminopeptidase (Aps) from Pseudomonas aeruginosa GF31 in B. subtilis, achieving an impressive enzyme activity of 13.7 U/mg. Notably, we have demonstrated, for the first time, the Aps-mediated degradation of diverse aromatic esters, including but not limited to pyrethroids, phthalates, and parabens. A biochemical characterization of Aps reveals its esterase properties and a broader spectrum of substrate profiles. The degradation rates of p-nitrobenzene esters (p-NB) with different side chain structures vary under the action of Aps, showing a preference for substrates with relatively longer alkyl side chains. The structure-dependent degradability aligns well with the binding energies between Aps and p-NB. Molecular docking and enzyme–substrate interaction elucidate that hydrogen bonding, hydrophobic interactions, and π–π stacking collectively stabilize the enzyme–substrate conformation, promoting substrate hydrolysis. These findings provide new insights into the enzymatic degradation of aromatic ester pollutants, laying a foundation for the further development and modification of promiscuous enzymes.
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Foods
·
2025
·Vol. 14
·期 17
·3055
·
IF 5.1
Xianxian Luo,Hao Chen,Jiayi Mi,Xinyan Li,Ziheng Wu,Yan Jiang,Xiufang Dong
Macroalgae are often used to produce sodium alginate, but their by-products have not been fully utilized. This study aimed to optimize the extraction of bound polyphenols (BPs) from Macrocystis pyrifera (L.) residues, analyze the composition of free polyphenols (FPs) and BPs, and evaluate their antioxidant activities and ability to inhibit glycosidase activity. The optimal conditions for extracting BPs achieved by the response surface method were as follows: 50 °C, a solid–liquid ratio of 1:50, an alkaline hydrolysis time of 2.38 h, and a NaOH concentration of 8 mol/L. Polyphenol content determination results indicated that FPs had significantly higher total polyphenols (13.02 ± 0.05 μg GAE/mg) and phlorotannin (3.44 ± 0.04 μg PE/mg) than BPs (6.57 ± 0.07 μg GAE/mg and 1.32 ± 0.20 μg PE/mg). HPLC/ESI-QTOF-MS showed distinct profiles: FPs had one polyhydroxy phenol, nine flavonoids, and four additional compounds, whereas BPs had five flavonoids and four other compounds. Antioxidant activity was found to be higher in FPs than in BPs (DPPH: 3.03 vs. 1.79 μg TE/mg; FRAP: 19.40 vs. 7.43 μg TE/mg). Furthermore, FPs exhibited 4.59- and 11-fold higher inhibition capacity toward α-amylase and α-glucosidase, respectively, compared to BPs. The results provide valuable basic data for the application of macroalgae residues in the marine biological industry and reveal their potential hypoglycemic ability.
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JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
·
2025
·Vol. 105
·期 7
·3931-3941
·
IF 3.5
Jiawei Zheng,Qiangyue Zhang,Nanjing Zhong
BACKGROUND Immobilized enzyme possessing both high activity and good selectivity is important in practice. In this study, Candida antarctica lipase B (CALB) was immobilized onto the macroporous resin ADS-17 for triacylglycerol (TAG) synthesis through esterification of oleic acid and glycerol. The reaction conditions were optimized by single-factor study and orthogonal test, and the reusability of the immobilized CALB (CALB@ADS-17) was evaluated. In addition, the mechanism of lipase immobilization was studied and the catalytic mechanism of CALB@ADS-17 was investigated. RESULTS Oleic acid conversion up to 99.20% and TAG content at 91.58 wt% could be obtained under optimal conditions. In addition, the CALB@ADS-17 retained 84.28% of its initial activity after 11 cycles of reuse. The mechanism of lipase immobilization was through hydrophobic adsorption. The relationship between temperature and oleic acid conversion was ln V 0 = 6.3316 − 4.3321/ T , and the activation energy ( E a ) was 36.02 kJ mol −1 . CALB@ADS-17 did not exhibit an obvious interfacial activation phenomenon. Its kinetic behavior can be described by the Michaelis–Menten model, whose kinetic parameters of v max , k cat , K m , K i , and k ca t /K m were 0.01265 μmol L −1 s −1 , 9310.72 s −1 , 0.4907 mmol L −1 , 3.997 mmol L −1 , and 1.90 × 10 4 L mmol −1 s −1 , respectively. CONCLUSION CALB@ADS-17 showed good esterification performance and exhibited good selectivity towards TAG generation. In addition, CALB@ADS-17 exhibited good reusability in esterification reactions and has potential in practical applications. © 2025 Society of Chemical Industry.
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