ACS Sustainable Chemistry & Engineering
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2022
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IF 9.224
Wen Tang,Haoxiang Li,Wei Zhang,Tonghao Ma,Jiafeng Zhuang,Ping Wang,Chao Chen
Site-specific and covalent attachment is the most desirable immobilization strategy, but classic methods typically require genetic engineering or complicated material fabrication, resulting in operational complexity and difficulty. Herein, a novel site-specific and covalent immobilization strategy based on accurately selected immobilization sites on lipase was developed. Specifically, computer-aided structural analysis of functional groups on lipase revealed that lysine residues with free amino groups were far away from the catalytic pocket and lid, which were suitable to be chosen as the best immobilization sites to effectively reduce the loss in its activity. Meanwhile, natural polyphenol-modified magnetic nanoparticles could increase the active immobilization sites, and lipase can be immobilized on them directly via a covalent reaction. This site-specific immobilization system exhibited significant enhancement in activity recovery (71.3%) compared to random immobilization (42.5 and 55.9%). As expected, experimental and computational analyses revealed that tailor-made site-specific immobilization carriers were beneficial to maintain the native catalytic pocket conformation and enhance the rigidity of the immobilized lipase, which exhibited a higher biodiesel yield (92.1%) than free and randomly immobilized lipases. Besides, the site-specifically immobilized lipase could maintain as high as 75.3% biodiesel yield after eight cycles, making it an ideal nanocatalyst for efficient production of biodiesel. Overall, the site-specific enzyme immobilization technology can provide stable catalytic activity advantages over the randomly covalent immobilization strategy, which can significantly promote green manufacturing and sustainable production.
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高影响因子
CHINESE JOURNAL OF CHEMICAL ENGINEERING
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2019
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IF 1.911
Ning Chen,Chunyu Zhang,Xiaoyan Dong,Yan Sun
Our previous work proved that the thermal stability of Candida rugosa lipase (CRL) immobilized on zwitterionic polymer (poly(carboxybetaine methacrylate)) grafted silica nanoparticle (SNP) was much higher than that on poly(glycidyl methecrylate) (pGMA) grafted SNP, while the latter showed significantly increased activity. Inspired by the research, we have herein proposed to synthesize copolymers of zwitterionic sulfobetaine methacrylate (SBMA) and GMA for CRL immobilization. The copolymers were grafted onto SNP surface at three GMA/SBMA (G/S) molar ratios (G100/S0, G50/S50, G10/S90), followed by the covalent coupling of CRL to the surface copolymers . The immobilized CRLs on the corresponding supports were denoted as p(G100-S0)-CRL, p(G50-S50)-CRL and p(G10-S90)-CRL. The enzyme loading increased with the increase of GMA content in the copolymer, while the activity varied with the grafted copolymer composition . Kinetic study proved the improvement of enzyme-substrate affinity after immobilization. In comparison to p(G100-S0)-CRL, p(G50-S50)-CRL and p(G10-S90)-CRL presented remarkably enhanced thermal stability and pH tolerance, and p(G10-S90)-CRL showed the highest stability. These results suggest that the copolymer design is promising for development as a versatile platform for enzyme immobilization .
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JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
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2024
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IF 3.3
Buqing Wang,Yasen Wang,Xiaoman Zhou,Xiao-Dong Gao,Morihisa Fujita,Zijie Li
BACKGROUND Acidic lipases with high catalytic activities under acidic conditions have important application values in the food, feed and pharmaceutical industries. However, the availability of acidic lipases is still the main obstacle to their industrial applications. Although a novel acidic lipase Rasamsonia emersonii (LIPR) was heterologously expressed in Escherichia coli , the expression level was unsatisfactory. RESULTS To achieve the high-efficiency expression and secretion of LIPR in Pichia pastoris GS115, the combinatorial optimization strategy was adopted including gene codon preference, signal peptide, molecular chaperone co-expression and disruption of vacuolar sorting receptor VPS10 . The activity of the combinatorial optimization engineered strain in a shake flask reached 1480 U mL −1 , which was 8.13 times greater than the P. pastoris GS115 parental strain. After high-density fermentation in a 5-L bioreactor, the highest enzyme activity reached as high as 11 820 U mL −1 . LIPR showed the highest activity at 40 °C and pH 4.0 in the presence of Ca 2+ ion. LIPR exhibited strong tolerance to methanol, indicating its potential application in biodiesel biosynthesis. Moreover, the gastrointestinal digestion simulation results demonstrated that LIPR was tolerant to pepsin and trypsin, but its activity was inhibited by sodium taurodeoxycholate. CONCLUSION This study provided an effective approach for the high expression of acidic lipase LIPR. LIPR was more appropriate for lipid digestion in the stomach than in intestine according to the gastrointestinal digestion simulation results. © 2024 Society of Chemical Industry.
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高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
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2015
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IF 2.297
Xuping Wang,Qingyun Tang,Grzegorz Maria Popowicz,Bo Yang,Yonghua Wang
More and more industrial chemistry reactions rely on green technologies. Enzymes are finding increasing use in diverse chemical processes. Epoxidized vegetable oils have recently found applications as plasticizers and additives for PVC production. We report here an unusual activity of the Malassezia globosa lipase (SMG1) that is able to catalyze epoxidation of alkenes. SMG1 catalyzes formation of peroxides from long chain carboxylic acids that subsequently react with double bonds of alkenes to produce epoxides. The SMG1 is selective towards carboxylic acids and active also as a mutant lacking hydrolase activity. Moreover we present previously unobserved mechanism of catalysis that does not rely on acyl–substrate complex nor tetrahedral intermediate. Since SMG1 lipase is activated by allosteric change upon binding to the lipophilic–hydrophilic phase interface we reason that it can be used to drive the epoxidation in the lipophilic phase exclusively.
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PubMed
高影响因子
MATERIALS SCIENCE FORUM
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2020
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IF 0
Hong Qian Dai,Yuan Lu,Zhi Hin Ou,Min Xu
Fe3O4-APTES-CS2 was prepared by modification of Fe3O4 with 3-aminopropyltriethoxysilane (APTES) and carbon disulfide (CS2). Lipase was covalently bonded to Fe3O4-APTES-CS2 to obtain the immobilized lipase Fe3O4-APTES-CS2-lipase. The textural characteristic of Fe3O4-APTES-CS2-lipase was assessed by scanning electron microscopy. The optimal immobilization conditions were 2.5 mg/mL lipase, pH 7.0, 35 °C, 3 h. The loading amount of lipase was 119.0 mg/g carrier. The immobilization efficiency reached 57%. (R)-(+)-N-acetyl-1-methyl-3-amphetamine was synthesized by Fe3O4-APTES-CS2-Li- pase. The conversion, enantiomeric excess of (R)-(+)-N-acetyl-1-methyl-3-amphet-a- mine and E value reached 36.8%, 98.5% and 132, respectivel.
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FOOD CHEMISTRY
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2020
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IF 6.306
Rui Huang,Yu Zhang,Shuyu Shen,Zijian Zhi,Huan Cheng,Shiguo Chen,Xingqian Ye
Obesity and oxidative damage are two important risk factors associated closely with metabolic syndrome. Utilization of functional food ingredients is considered as a feasible way to tackle these challenges. In the present study, eight representative species of citrus peel extracts (CPEs) were evaluated and compared for their flavonoid profiles, antioxidant activities, and pancreatic lipase (PL) inhibitory capacities and mechanisms. Results indicated that hesperidin, naringin, neohesperidin, narirutin and eriocitrin were the five major flavonoids in CPEs, among which hesperidin was the main active PL inhibitor. Moreover, hesperidin could interact with PL by hydrogen bonds and van der Waals forces , and the interaction would not obviously change the secondary structure of PL. Overall, ponkan peel extract, having the strongest overall antioxidant activity, the highest content of hesperidin and total phenolic compounds among all tested CPEs, is a promising natural ingredient to scavenge free radicals and manage obesity.
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高影响因子
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
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2024
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IF 3.3
Chunqi Lu,Qiuya Gu,Xiaobin Yu
BACKGROUND Fucoidan has an anti-obesity effect. However, there are few studies on its mechanism. In this study, we investigated the in vitro and in silico inhibitory properties of fucoidan against pancreatic lipase for the first time. We examined the changes in composition, structure, and pancreatic lipase inhibition of fucoidan during in vitro digestion. RESULTS Simulated saliva-gastrointestinal digestion resulted in a slight decrease in the molecular weight of fucoidan but no significant changes in the monosaccharide composition, sulfate content, and functional groups. Moreover, the digestion process significantly increased the inhibition of pancreatic lipase by fucoidan. The study on the type of inhibition showed that the inhibition of pancreatic lipase by fucoidan belonged to mixed inhibition with competitive inhibition. Molecular docking analysis showed that fucoidan could bind to the active site of pancreatic lipase. CONCLUSION This study indicates that fucoidan can be a potential functional food for anti-obesity. © 2024 Society of Chemical Industry.
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高影响因子
Advanced Healthcare Materials
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2026
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IF 11
Dongqiu Li,Bingyi Xu,Feifei Ren,Jiajing Yu,Yan Tong,Shiwu Deng,Hezhong Jiang
Diabetic wound healing is substantially impaired by biofilm infections, oxidative stress, and persistent hypoxia, which present major challenges for timely diagnosis and treatment. In this study, theranostic nanoparticles (NPs) were engineered to facilitate lipase-triggered biofilm theranostics and accelerate wound healing. Theranostic Mn-TC NPs were prepared by grafting a fluorescent sonosensitizer, meso-tetra (4-carboxyphenyl) porphine (TCPP), onto manganese dioxide (MnO 2 ) nanoflowers, quenching the fluorescence emissions of TCPP. Upon encountering biofilms in vivo, the elevated lipase hydrolyzes ester linkages within the Mn-TC NPs, liberating TCPP to restore its fluorescence emission and enabling the real-time visualization of biofilm-infected wounds. MnO 2 nanoflowers offer abundant reaction sites for TCPP grafting while enhancing the catalysis of hydrogen peroxide to generate oxygen. The boosted oxygen evolution promoted the sonodynamic therapy effect of ultrasound-activated TCPP, achieving 94.0% reduction in biofilm biomass and 99.9% bacterial clearance. Engineering NPs accelerate wound healing by simultaneously eradicating biofilms, modulating inflammatory states, enhancing collagen deposition, and promoting angiogenesis. This study presents a novel theranostic strategy for biofilm-triggered visual imaging and an antibiotic-free therapy for diabetic wounds.
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高影响因子
INDUSTRIAL CROPS AND PRODUCTS
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2026
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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
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2026
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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
BIORESOURCE TECHNOLOGY
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2026
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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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高影响因子
Biomed Research International
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2015
·Vol. 2015
·期 1
·139179
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IF 1.579
Jingjing Sun,Yiling Chen,Jun Sheng,Mi Sun
To improve the reusability and organic solvent tolerance of microbial lipase and expand the application of lipase (hydrolysis, esterification, and transesterification), we immobilized marine microbial lipase using different methods and determined the properties of immobilized lipases. Considering the activity and cost of immobilized lipase, the concentration of lipase was fixed at 2 mg/mL. The optimal temperature of immobilized lipases was 40°C and 5°C higher than free lipase. The activities of immobilized lipases were much higher than free lipase at alkaline pH (more than 50% at pH 12). The free lipase lost most activity (35.3%) and immobilized lipases retained more than 46.4% of their initial activity after 3 h heat treatment at 70°C. At alkaline pH, immobilized lipases were more stable than free lipase (more than 60% residue activity at pH 11 for 3 h). Immobilized lipases retained 80% of their activity after 5 cycles and increased enzyme activity (more than 108.7%) after 3 h treatment in tert-butanol. Immobilization of lipase which improved reusability of lipase and provided a chance to expand the application of marine microbial lipase in organic system expanded the application range of lipase to catalyze hydrolysis and esterification in harsh condition.
研究领域:
酶工程
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分子生物学
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生物催化
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工业微生物学
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蛋白质化学
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应用酶学
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危险品化学品经营许可证（带存储）