LANGMUIR
·
2026
·
IF 4.4
Xinyi Wu,Siqi Zhu,Siqi Zheng,Guoxiang Wang,Yong Gao
Enzymatic microreactors with sustainable operational stability have attracted significant interest in the development of green synthesis. In this work, a recyclable and pH-responsive enzymatic microreactor was constructed on the basis of high internal phase Pickering emulsions (HIPPEs). Oil-in-water HIPPEs were prepared via simple hand-shaking emulsification adopting amine-functionalized diblock copolymer nanoparticle/lipase complexes as Pickering emulsifiers. The amine-functionalized nanoparticles, composed of poly(oligo(ethylene glycol) methyl ether methacrylate)-b-poly(diacetone acrylamide), were synthesized through the combination of RAFT-mediated polymerization-induced self-assembly and subsequent hydrolysis. The obtained HIPPEs displayed reversible emulsification/demulsification behaviors triggered by protonation/deprotonation of amine groups on the surfaces of nanoparticles. When applied to the esterification of butyric acid with 1-butanol, the as-prepared HIPPE-based microreactor exhibited outstanding catalytic efficiency due to the large interfacial area and shortened diffusion distance provided by the high-internal-phase structure. Moreover, pH-triggered demulsification enabled convenient product separation and efficient recycling of the nanoparticle/lipase complexes for repeated catalytic runs, demonstrating good sustainability and promising potential for practical applications.
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BIORESOURCE TECHNOLOGY
·
2026
·
IF 8.2
Zihan Zhang,Yu Zhang,Qingqing Liu,Wanqing Song,Yi Hu,Binbin Nian
A novel triple evolution strategy overcame the lipase activity-stability trade-off. • CM5 showed 2.3-fold higher activity and 4.8-fold solvent tolerance than WT. • MD simulations link CM5 stability to rigidity and stable hydration shell. • The VES synthesis strategy achieved the highest yield and reusability to date. • A complete chain from computational design to practical production was validated. Biocatalysts exhibit broad application prospects in industrial production. However, the inherent instability of natural enzymes in non-aqueous media severely limits their practical utilization. Using Candida rugosa lipase as a model, a synergistic evolution strategy integrating ancestral sequence reconstruction, free energy optimization and surface hydration shell modulation was developed in this study. The optimal mutant CM5 exhibited simultaneously enhanced stability and activity, showing a 4.78-fold increase in tolerance to 65% (v/v) dimethyl sulfoxide, a 1.67-fold extension in half-life at 50 ℃ and a 2.30-fold increase in activity compared to the wild type. Molecular dynamics simulations indicated that the enhanced stability of CM5 originates from improved structural rigidity and the stabilized hydration shell. Meanwhile, its increased activity is attributed to a reduced energy barrier for the lid conformational transition and an optimal enzyme substrate affinity that significantly boosts substrate turnover efficiency. Moreover, a volumetric activity of 142.71 U/mL in a 5 L bioreactor was obtained via optimization of the chassis cells, and the resulting culture was then used for the immobilization. The lipase CM5 immobilized on amino octadecyl bifunctional macroporous resin (CM5@AOMR) achieved a vitamin E succinate yield of > 99.9% and maintained an outstanding yield of 98.2% even after ten consecutive batch reactions. Overall, this study developed a comprehensive workflow from computational design and efficient fermentation to immobilization and practical applications of lipase, providing a promising new approach for developing next generation biocatalysts suited for extreme industrial environments. Download: Download high-res image (234KB) Download: Download full-size image
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INDUSTRIAL CROPS AND PRODUCTS
·
2026
·
IF 6.4
Wei Wu,Yunzhu Chen,Lijuan Jiang,Peiwang Li,Yan Yang,Wenyan Yuan,Zhihong Xiao,Xiao Zhou,Qiang Liu,Wenbin Zeng,Luhong Zhang,Jingzhen Chen,Changzhu Li
SHSL was mined from Streptomyces sp. CL10 and expressed as an alkaline lipase. • LC-MS showed TAG depletion, DAG enrichment, and low MAG in Camellia oleifera oil. • Triolein, diolein, and monoolein were used to model acylglycerol recognition. • Docking showed substrate-dependent positioning in the SHSL catalytic groove. • MD revealed triolein-favored catalytic geometry for TAG-to-DAG redistribution. Enzymatic modification of woody plant oils can improve their functional glyceride composition and expand their value in lipid-based applications. In this study, a hormone-sensitive lipase, SHSL, was identified from Streptomyces sp. CL10 through genome mining and domain/motif-based screening. Recombinant SHSL showed alkaline lipase activity, with an optimal temperature of 45°C and an optimal pH of 8.0. LC-MS analysis showed that SHSL treatment reshaped the detected glyceride pool of Camellia oleifera oil, characterized by TAG depletion, DAG enrichment, and low MAG levels. Docking and 120-ns MD simulations indicated that SHSL accommodated oleoyl-rich TAG in a more favorable catalytic geometry than downstream DAG and MAG substrates, involving Ser150 proximity and oxyanion-hole stabilization. These results provide a structural interpretation for TAG-to-DAG-biased redistribution and support SHSL as a candidate biocatalyst for enzymatic modification of C. oleifera oil. Download: Download high-res image (212KB) Download: Download full-size image
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Food Bioscience
·
2026
·
IF 6.2
Lunaike Zhao,Yun Huang,Yanmei Xi,Weizhe Wang,Yufang Su,Baoguo Sun,Nasi Ai
Lipase supplementation improves the flavor profile of UHT milk during storage. • Methyl ketones constitute the primary source of off-flavors in UHT milk during late storage. • Lactones endow UHT milk with enhanced milky and sweet aroma. • Lipase improves milk flavor mainly within the first three months of storage. Lipase-mediated flavor improvement in dairy products has been widely investigated. However, the majority of previous studies have focus on flavor attributes upon completion of enzymatic hydrolysis, with limited attention has been paid to the dynamic flavor changes of lipase-supplemented UHT milk during storage. This study comprehensively investigated the effects of lipase treatment (0, 0.1, 1, and 10 U/L) on quality properties, color variations, sensory profiles, and volatile compound profiles of UHT milk throughout six months of storage. Lipase supplementation reduced the pH value and fat content of UHT milk and shifted its color toward pale yellow. Sensory evaluation revealed that lipase significantly enhanced the milky, sweetness, and fatty flavor attributes of UHT milk within the first three months of storage, while the overall acceptability was also improved during this period. Based on volatile profiling, PLS-DA model and relative odor activity values (ROAV), 2-heptanone, 2-nonanone, 2-undecanone, octanoic acid, hexanoic acid, 1-octanol, 2-furanmethanol, delta-dodecalactone, delta-decalactone, pentanal, and benzaldehyde were identified as key volatile compounds associated with lipolytic changes in UHT milk. Integrating the aromatic characteristics of volatile compounds and dynamic sensory changes indicated that methyl ketones and short-chain fatty acids exert dual effects on aroma enhancement and off-flavor formation in UHT milk, whereas lactones contribute to improving the sensory properties of milk flavor. This study demonstrates the potential of exogenous lipase supplementation to modulate UHT milk flavor, and reveals the patterns of lipase-associated dynamic changes in volatile flavor compounds of UHT milk during storage. Download: Download high-res image (371KB) Download: Download full-size image
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高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2023
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IF 8.2
Yan-Wei Liu,Qing-Hao Li,Shi-Yu Li,Guo-Qing Huang,Jun-Xia Xiao
To prompt the application of the chitosan (CS)- Aspergillus oryzae lipase (AOL) complex in the construction of novel biphasic catalysis medium, its Pickering emulsion stabilization ability as well as adsorption behavior in the oil-water interface were investigated and the stability of resultant emulsion was evaluated. The results indicated that the CS-AOL complex assembled in mass ratio 1:5 was an effective Pickering stabilizer and up to 90 % AOL could be retained in the emulsion interface. Quartz crystal microbalance with dissipation monitoring suggested that the CS-AOL complex spontaneously absorbed to oil-water interface; absorption dynamics analysis revealed that the adsorption was driven by diffusion accompanied by rapid structural rearrangement; while interfacial dilatational rheology demonstrated the formation of an elastic film in the oil-water interface. The Pickering emulsions were pseudoplastic and that in oil fraction 0.6 exhibited the elastic behavior in contrast to the viscous behavior in oil fractions 0.2 and 0.4. The Pickering emulsion exhibited excellent stability against storage for up to 28 d, pHs 2.0–12.0, heating at 25–90 °C, and up to 500 mmol/L NaCl, and the corresponding interfacial AOL retentions exceeded 80 % during exposure to these conditions. Hence, the CS-AOL complex could be used as a stabilizer to construct Pickering emulsion-based biphasic catalysis systems.
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PROCESS BIOCHEMISTRY
·
2022
·
IF 4.885
Zexin Zhao,Jiahao Huang,Long Xu,Changgao Wang,Jun Cai
The efficient methanol-resistant biocatalysts with low costs are always desirable for biodiesel production. Here, an Escherichia coli whole-cell catalyst was constructed with the methanol-resistant MAS1 lipase being expressed intracellularly. Notably, this whole-cell catalyst showed significantly higher methanol-resistance than the purified MAS1 lipase. The whole-cell catalyst with the non-specific MAS1 lipase enabled efficient biodiesel production. Moreover, high-density fermentation was found to reduce the production costs and improve the catalytic activity of the whole-cell catalyst. The wet cells collected by centrifugation were directly used for fatty acid methyl ester (FAME) production from olive oil with one-step addition of methanol. Under the optimal conditions (catalyst loading of only 1.5 wt%, water content of 30 % (v/w), methanol content of 30 % (v/w) and reaction temperature of 29 °C), the maximum yield of 93.56 % was obtained. Furthermore, the whole-cell catalyst showed favorable reusability, with 85 % of the original activities left after a 4-cycle usage. Overall, the whole-cell catalyst reported in the present study would provide great potentials for the enzymatic production of biodiesels.
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APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2022
·Vol. 195
·期 2
·1216-1230
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IF 3.094
Cheng Qiaojuan,Chi Xinyu,Liang Yingchao,Li Wanxin,Sun Jiaxin,Tao Jin,Wang Zhi
Metal–organic frameworks (MOFs) can be used as the immobilization carriers to protect the physicochemical properties of enzymes and improve their catalytic performance. Herein, we report an in situ co-precipitation method to immobilize lipase from Candida sp. 99–125 in Cu-BTC MOF (BTC = 1, 3, 5-benzene tricarboxylic acid, H 3 BTC). Characterizations of the immobilized lipase (lipase@Cu-BTC) have confirmed the entrapment of lipase molecules in Cu-BTC MOF. The immobilized lipase has been successfully applied for resolving N- hydroxymethyl vince lactam ( N -HMVL) and its catalytic activity is five times that of native enzyme. More importantly, we found that Cu-BTC MOF can afford powerful protection for enzyme in nearly dry organic solvent and endow the immobilized lipase with excellent reusability and storage stability. Our present study may widen the application of immobilized enzyme with MOF as the immobilized carrier. Graphical Abstract
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Food Bioscience
·
2021
·
IF 4.24
Fuqiang Liang,Yumeng Shi,Weiwei Cao,Jiayi Shi
This study aimed to elucidate the inhibition mechanism of apigenin against porcine pancreatic lipase (PPL), and, moreover, to comprehensively reveal the molecular basis of its anti-obesity via network pharmacology approach. The results showed that apigenin inhibited PPL with an IC 50 value of 0.377 ± 0.041 mM. Spectroscopic techniques combined molecular docking suggested that apigenin could bind into the PPL active pocket, affecting its normal spatial conformation. Moreover, molecular dynamic (MD) simulations revealed that the open conformation of PPL tended to transit to the closed in the presence of apigenin, which might one important reason for the inhibition of PPL catalytic ability. Network pharmacology analysis revealed that a total of 49 proteins could be identified as potential targets for the anti-obesity effects of apigenin. According to the protein-protein interaction (PPI) network analysis, six hub targets were extracted including IGF1 , ESR1, MMP9 , PPARA , MAPK14 and NR3C1. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment indicated that the 49 potential targets could be mapped to 30 pathways (p<0.05). Among them, PI3K-Akt signaling pathway and insulin resistance can be considered as two major pathways regulated by apigenin. Further docking studies indicated that apigenin can bind into the binding pocket of the six hub target proteins identified according to the PPI network. The results indicated that in addition to inhibiting PPL, apigenin could exhibit anti-obesity benefit through the molecular mechanisms uncovered by network pharmacology. This study proposes a new strategy to reveal the mechanisms of dietary polyphenols at the level of network pharmacology.
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JOURNAL OF MOLECULAR LIQUIDS
·
2021
·
IF 6.165
Sha Li,Xing Hu,Junhui Pan,Deming Gong,Guowen Zhang
The inhibition of pancreatic lipase by apigenin, a natural dietary flavonoid, was investigated by various spectroscopic techniques and computational simulation. The results showed that apigenin reversibly inhibited pancreatic lipase activity in a competitive manner with an IC 50 value of 0.45 ± 0.03 mM, and its combination with orlistat showed a synergistic effect. Formation of a complex between apigenin and pancreatic lipase driven by hydrogen bonds resulted in the fluorescence quenching of the enzyme through a static manner. The binding constant of apigenin with the enzyme was (5.47 ± 0.01) × 10 4 L mol −1 at 298 K, and the kinetic process of their interaction was characterized by the resolved concentration profiles and pure spectra of different components from the multivariate curve resolution-alternating least squares algorithm. The inhibition of pancreatic lipase by apigenin was mainly attributed to the tighter structure of the enzyme and the interaction between apigenin and the amino acid residues in the active center of the enzyme to prevent the substrate from being oxidized, as demonstrated by the analysis of circular dichroism spectra and molecular docking. Molecular dynamics simulation further characterized the binding conformation and property of apigenin-pancreatic lipase complex. This study provides novel insights into the mechanism of apigenin as pancreatic lipase inhibitor and the development of apigenin as an anti-obesity nutrient.
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JOURNAL OF MOLECULAR LIQUIDS
·
2020
·
IF 5.065
Rongrong Wang,Wei Jin,Xirong Huang
In the present work, the sulfobetaine-type zwitterionic surfactant (SB- n ) was tried for the first time to construct hydrophobic ionic liquid (HIL)-based bicontinuous microemulsions . The composition-dependent phase behavior of the HIL/water (buffer)/SB- n ternary microemulsion system was determined by the method of T - γ fishlike phase diagram . It is found that the phase inversion temperature increases markedly with the increase of the alkyl chain length ( n ) of SB- n , but the surfactant efficiency changes little. The HIL greatly affects the phase behavior, depending on the ion structure. It is found that the SB-12 stabilized [C 8 mim][PF 6 ]-based microemulsion system has moderate phase inversion temperature, and moreover, its surfactant efficiency is the highest among the reported HIL-based bicontinuous microemulsions stabilized by conventional surfactants. The composition-dependent microstructure of the SB-12-stabilized [C 8 mim][PF 6 ]-based bicontinuous microemulsion was characterized by SAXS technique, and its influence on the catalytic performance of solubilized lipase was also explored. With the increase of the mass ratio of the HIL to water ( α ), the correlation length ( ξ ) changes slightly, but the microdomain size ( d ) decreases steadily. Unlike α , with the increase of the surfactant concentration ( γ ), the d changes little, but the ξ increases markedly. This abnormal change of ξ with γ should be ascribed to the interface rigidity of the microemulsion. The catalytic efficiency of lipase depends on the rigidity of the interface and its interfacial area. The above original studies not only enrich HIL-based microemulsion systems, but also provide scientific support for lipase-based green synthesis and transformation.
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COLLOIDS AND SURFACES B-BIOINTERFACES
·
2020
·
IF 4.389
Qing Li,Yingxuan Chen,Shaowei Bai,Xinxin Shao,Lin Jiang,Quanshun Li
Herein, thermophilic lipase QLM from Alcaligenes sp. has been successfully immobilized in bio-based metal-organic frameworks (MOFs) through biomimetic mineralization, using zinc acetate and adenine as metal ion and organic ligand, respectively. The morphology and structure of lipase@Bio-MOF was systematically characterized by scanning electron microcopy (SEM), transmission electron microcopy (TEM), powder X-ray diffraction (PXRD) and Fourier transform infrared spectra (FT-IR). The enzyme loading in immobilized enzyme was measured to be 15.9 % by thermogravimetric analysis (TGA). Further, it was demonstrated to possess favorable catalytic activity and stability under high temperature and alkaline conditions and in the presence of metal ions, using the hydrolysis of p -nitrophenyl caprylate as a model. Finally, the immobilized enzyme was successfully applied in the preparation of biodiesel through the trans-esterification of sunflower oil with methanol, obtaining a conversion of >60 % at a high oil/methanol ratio of 8:1. Meanwhile, it showed excellent recyclability during the biodiesel production, and no changes of morphology and crystal structure were observed after being used for 3 cycles. Overall, the immobilized lipase in bio-based MOFs provided an economically and environmentally viable biocatalyst for the synthesis of biodiesel.
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APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2020
·Vol. 192
·期 2
·392-414
·
IF 2.277
Chen Sunni,Li Jing,Fu Zejian,Wei Guohua,Li Hongyan,Zhang Bin,Zheng Liufeng,Deng Zeyuan
Phytosterols are regarded as compounds able to reduce total and low-density lipoprotein cholesterol in the blood, and their esterified derivatives could help to improve the effectiveness of this function. In the present study, the water/sodium 1,4-bis-2-ethylhexylsulfosuccinate (AOT)/isooctane reverse micelle (RM) system was set up as a reaction medium for Candida rugosa lipase AY30 (CRL AY30) to synthesize β-sitosterol laurate (β-SLE). The product was identified by TLC, FT-IR, and HPLC–APCI–QqQ–MS/MS and quantified by HPLC. Through stepwise optimization, it was found that CRL AY30 had the highest activity in the water/AOT/isooctane RM system where 50 mM PBS with a pH of 7.5 was adopted as water core to carry CRL AY30, and the proportion of [CRL AY30] (mg/mL), [water] (mM), and [AOT] (mM) was set in 3:375:25, respectively, in isooctane. After screened with single-factor experiments, the esterification reaction conditions in the CRL AY30–water/AOT/isooctane RM system were further optimized by the response surface method as follows: the mole ratio of β-sitosterol to lauric acid of 1:3.5 (25 mM β-sitosterol), the enzyme load of 18% (w/w total reactants), the reaction temperature of 47 °C, and the reaction time of 48 h. As a result, the maximum esterification rate was up to 88.12 ± 0.79%.
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危险品化学品经营许可证（带存储）