JOURNAL OF BIOTECHNOLOGY
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2018
·
IF 2.533
Binglin Li,Dandan Duan,Jiao Wang,Huanyu Li,Xiaoli Zhang,Binxia Zhao
Phospholipase D (PLD) was firstly hyperactivated by the bio-imprinting and, then, these hyperactivated PLD molecules were immobilized by adsorption and precipitation, followed by cross-linking. The high degree of conformational rigidification provided by intra- and intermolecular cross-linking can make PLD “remember” imprint-induced characteristics even in aqueous solutions. The obtained immobilized PLD showed the excellent catalytic performance. The maximum activity of immobilized PLD reached 166953 U/g protein , which was approximately 14 times higher than that of free form (11922 U/g protein ). Moreover, the selectivity of PLD was significantly enhanced after immobilization. The yield of PG and PA reached 94.0% and 5.96%, respectively. Compared with the serious hydrolysis in the free PLD (35.3% yield of PA), the side reaction was minimized in this work. This may be the first description of the remarkably high improvement of the activity and selectivity of PLD through the immobilization technology.
研究领域:
生物化学
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酶工程
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DOI
PubMed
高影响因子
MOLECULES
·
2015
·Vol. 20
·期 4
·6273-6283
·
IF 2.416
Yun-Feng Zheng,Juan-Hua Wei,Shi-Qi Fang,Yu-Ping Tang,Hai-Bo Cheng,Tian-Lin Wang,Cun-Yu Li,Guo-Ping Peng
Two novel oleanane-type triterpene saponins, licorice-saponin P2 (1) and licorice-saponin Q2 (3), together with nine known compounds2,4–11, have been isolated from the water extract of the roots ofGlycyrrhiza inflata. The structures of these compounds were elucidated on the basis of spectroscopic analysis, including 2D-NMR experiments (1H–1H COSY, HSQC, HMBC and ROESY). Inin vitroassays, compounds2–4,6and11showed significant hepatoprotective activities by lowering the ALT and AST levels in primary rat hepatocytes injured by D-galactosamine (D-GalN). In addition, compounds2–4,6,7and11were found to inhibit the activity of PLA2with IC50values of 6.9 μM, 3.6 μM, 16.9 μM, 27.1 μM, 32.2 μM and 9.3 μM, respectively, which might be involved in the regulation of the hepatoprotective activities observed.Keywords:Glycyrrhiza inflate;genusGlycyrrhiza;triterpene saponin;hepatoprotective;phospholipase A2
研究领域:
药用植物化学
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肝脏保护研究
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PubMed
高影响因子
Biomolecules
·
2020
·Vol. 10
·期 2
·231
·
IF 4.082
Rabia Durrani,Faez Iqbal Khan,Shahid Ali,Yonghua Wang,Bo Yang
Phospholipase B (EC 3.1.1.5) are a distinctive group of enzymes that catalyzes the hydrolysis of fatty acids esterified at thesn-1andsn-2positions forming free fatty acids and lysophospholipids. The structural information and catalytic mechanism of phospholipase B are still not clear. Herein, we reported a putative phospholipase B (TmPLB1) fromTalaromyces marneffeiGD-0079 synthesized by genome mining library. The gene (TmPlb1) was expressed and the TmPLB1 was purified usingE. colishuffle T7 expression system. The putative TmPLB1 was purified by affinity chromatography with a yield of 13.5%. The TmPLB1 showed optimum activity at 35 °C and pH 7.0. The TmPLB1 showed enzymatic activity using Lecithin (soybean > 98% pure), and the hydrolysis of TmPLB1 by31P NMR showed phosphatidylcholine (PC) as a major phospholipid along with lyso-phospholipids (1-LPC and 2-LPC) and some minor phospholipids. The molecular modeling studies indicate that its active site pocket contains Ser125, Asp183 and His215 as the catalytic triad. The structure dynamics and simulations results explained the conformational changes associated with different environmental conditions. This is the first report on biochemical characterization and structure dynamics of TmPLB1 enzyme. The present study could be helpful to utilize TmPLB1 in food industry for the determination of food components containing phosphorus. Additionally, such enzyme could also be useful in Industry for the modifications of phospholipids.Keywords:free fatty acids (FFAs);NMR (nuclear magnetic resonance);phospholipase B;Talaromyces marneffei;affinity chromatography
研究领域:
生物化学
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分子生物学
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食品科学
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DOI
PubMed
高影响因子
Catalysts
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2020
·Vol. 10
·期 9
·1057
·
IF 3.52
Jing Wang,Sheng Xu,Yang Pang,Xin Wang,Kequan Chen,Pingkai Ouyang
To achieve efficient bio-production of phospholipase D (PLD), PLDs from different organisms were expressed inE.coli. An efficient secretory expression system was thereby developed for PLD. First, PLDs fromStreptomycesPMF andStreptomyces racemochromogeneswere separately over-expressed inE.colito compare their transphosphatidylation activity based on the synthesis of phosphatidylserine (PS), and PLDPMFwas determined to have higher activity. To further improve PLDPMFsynthesis, a secretory expression system suitable for PLDPMFwas constructed and optimized with different signal peptides. The highest secretory efficiency was observed when the PLD * (PLDPMFwith the native signal peptide Nat removed) was expressed fused with the fusion signal peptide PelB-Nat inE. coli. The fermentation conditions were also investigated to increase the production of recombinant PLD and 10.5 U/mL PLD was ultimately obtained under the optimized conditions. For the application of recombinant PLD to PS synthesis, the PLD properties were characterized and 30.2 g/L of PS was produced after 24 h of bioconversion when 50 g/L phosphatidylcholine (PC) was added.Keywords:phospholipase D;phosphatidylserine;secretory expression;enzymatic catalysis
研究领域:
生物化学
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分子生物学
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DOI
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2024
·
IF 3.7
Qian Zhou,Neng-Bing Long,Rui-Feng Zhang
A new epoxy-based polymerizing/compositing synchronic process was carried out in 3D structure of a melamine sponge. The cured polymer formed a porous and wave-shaped coating on the surface of skeleton in a uniform way. The obtained composite sponge was used as a microflow support for the immobilization of phospholipase A 1 (PLA 1 , from Thermomyces lanuginosus/Fusarium oxysporum ) by co-adsorption/cross-linking with polyethylenimine (PEI). The loading amount and the specific activity of immobilized PLA 1 under the optimal immobilization conditions were 45.7 mg/g support and 9.1 U/mg protein . The PLA 1 -immobilized microflow supports were placed in a columnar reactor, in which transesterification was operated in continuous circulation way. The reaction produced DHA/EPA-incorporated phosphatidylcholine , the hydrolysis side reaction could be inhibited by removal of free water in the reaction medium. Under optimized conditions the incorporation percentage of DHA/EPA and PC yield of 54.3% and 82.4% within 18 h at 55 ℃, respectively. After 10 cycles, the immobilized PLA 1 retained 71.2% of the original activity. The use of microflow support has an advantage on easier scaling up the reactor size towards much larger productivity and higher efficiency.
研究领域:
生物化学
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酶工程
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DOI
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Zhong Li,Huilin Zhou,Xuan Liu,Weifei Wang,Dongming Lan,Yonghua Wang
The limited availability of phospholipase A 1 (PLA 1 ) has posed significant challenges in enzymatic degumming. In this study, a novel PLA1 (UM2) was introduced to address this limitation, which had a unique thermo-responsive ability to switch phospholipase and lipase activities in response to temperature variations. Remarkably, UM2 displayed an unprecedented selectivity under optimized conditions, preferentially hydrolyzing phospholipids over triacylglycerols—a specificity superior to that of commercial PLA 1 . Moreover, UM2 demonstrated high efficiency in hydrolyzing phospholipids with a predilection for phosphatidylcholine (PC) and phosphatidylethanolamine (PE). A practical application of UM2 on crude flaxseed oil led to a dramatic reduction in phosphorus content, plummeting from an initial 384.06 mg/kg to 4.38 mg/kg. Broadening its industrial applicability, UM2 effectively performed enzymatic degumming for other distinct crude vegetable oils with a unique phospholipid composition. Collectively, these results highlighted the promising application of UM2 in the field of oil degumming.
研究领域:
生物化学
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食品科学
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DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
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2025
·
IF 3.3
He Ying,Huang Ao,Liu Yun
Phospholipase D (PLD) is a valuable enzyme in industrial processes for converting phosphatidylcholine (PC) to phosphatidylserine (PS). In this study, a strain of Bacillus cereus was isolated from soil and identified through 16S rRNA sequencing. To enhance PLD activity, various mutagenesis strategies—including chemical treatment, irradiation, and their combinations—were employed, resulting in four high-activity positive mutants (C-7, I-12, CI 7–12, and IC 13–14). Among these, the CI 7–12 mutant exhibited a significantly higher enzymatic activity, showing a 3.12-fold increase (312.2%) compared to the wild-type strain. Fermentation conditions were optimied using response surface methodology (RSM), achieving a PLD activity of 35 U/mL. The enzyme demonstrated stability over a wide temperature range (30–60 °C) and pH range (6–10), with a half-life of 128 days. Kinetic analysis revealed a V max of 20.04 μmol/h and a K m of 7.13 μmol/mL, indicating efficient activity. In bioconversion experiments, the PLD-enriched fermentation broth catalyzed the conversion of PC to PS, achieving a 53.0% conversion rate and a 92.3% selectivity for PS in a two-phase system. These findings expand the potential sources of PLD and underscore its applicability for PS production in biotechnological applications.
研究领域:
微生物学
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生物化学
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工业生物技术
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PubMed
高影响因子
3 Biotech
·
2021
·Vol. 11
·期 4
·1-9
·
IF 2.406
Ren Runxian,Zhai Lixin,Tian Qiaopeng,Meng Di,Guan Zhengbin,Cai Yujie,Liao Xiangru
Diphenyl phosphate (DPHP) has been increasingly detected in environmental samples, posing a potential hazard to humans and other organisms and arousing concern regarding its adverse effects. Biological degradation of DPHP is considered a promising and environmentally friendly method for its removal. In this study, the bagdpd gene was mined from the Bacillus altitudinis W3 genome and identified as a glycerophosphodiester phosphodiesterase by bioinformatics analysis. The enzyme was expressed and its biochemical properties were studied. When using bis(4-nitrophenyl) phosphate as substrate, enzyme activity was optimal at 55 °C and a pH of 8.5. The enzyme remained stable in the pH range of 8.0 − 10.0. The rBaGDPD enzyme degraded DPHP and the reaction product was identified as phenyl phosphate by LC–MS. This is the first report of a glycerophosphodiester phosphodiesterase exhibiting hydrolytic activity against DPHP. This study demonstrated that rBaGDPD could have the potential for bioremediation and industrial applications.
研究领域:
环境科学
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微生物学
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生物化学
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DOI
PubMed
高影响因子
Foods
·
2023
·Vol. 12
·期 6
·1289
·
IF 5.2
Xiao-Yan Liu,Wei Chen,Cheng-Tao Wang
Egg white (EW) is frequently used in bakery products because of its excellent foaming capabilities. However, egg yolk (EY) contamination often degrades the foaming characteristics of EW. The purpose of this study was to investigate the effect of different concentrations of phospholipase A1 (PLPA1) and lipase (LP) on EW. The changes in particle size distribution and potential before and after enzymatic digestion of EW with contaminated 0.5 wt% and 1.0%wt EY were tested. The foaming rate and foam stability were measured after the dispersions were digested with different concentrations of PLPA1 and LP. Additionally, the dispersion samples were used to prepare batter and angel cake, and the modulus, density, and microstructure of the batter were analyzed. Results showed that the potential absolute value increased when the EY was hydrolyzed by PLPA1. The distribution of yolk particle size showed a new aggregation and the average particle size decreased after LP hydrolysis. The dispersion samples hydrolyzed by PLPA1 and LP recovered all the properties of the samples at enzymatic concentrations of 500 U/g and 2500 U/g. This may be attributed to the changes in yolk particles resulting from the enzymatic digestion of EY and the production of amphiphilic lysophospholipids, fatty acids, and glycerol.Keywords:egg white;foaming properties;yolk-contaminated;lipase enzymolysis
研究领域:
食品科学
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化学
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PubMed
高影响因子
INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH
·
2016
·
IF 2.567
Binglin Li,Jiao Wang,Xiaoli Zhang,Binxia Zhao
Phospholipase D (PLD) was immobilized in a simple and effective way by adsorption and precipitation of the enzyme, followed by chemical cross-linking to form an “enzyme net” covering the surface of nonporous silicon dioxide nanoparticles. For catalyzing transphosphatidylation to produce phosphatidylethanolamine (PE), at pH 6.0 and 35 °C (the optimum operational conditions), the specific activity of immobilized PLD reached 15872 U/gprotein, which was approximately 1.15 times higher than the maximum value of specific activity of free PLD (13813 U/gprotein). A kinetic study demonstrates immobilized PLD had increases in catalytic activity and enzyme–substrate affinity. In addition, the thermostability and pH tolerance were significantly enhanced compared with free PLD. The half-life of immobilized PLD was significantly increased from 30 to 70 days at 4 °C (approximately 2.3 times). This novel method has been proven to be suitable for the production of robust biocatalysts.
研究领域:
生物化学
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纳米生物学
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DOI
高影响因子
JOURNAL OF FOOD SCIENCE AND TECHNOLOGY-MYSORE
·
2021
·Vol. 59
·期 1
·317-326
·
IF 2.701
Weining Wang,Tang Honglin,Chen Yan,Liu Jingyang,Yu Dianyu,Yang Fuming,Elfalleh Walid
In this paper, the free Phospholipase A 1 (PLA 1 ) was immobilized on a magnetic carrier. The average particle diameter of the magnetic carrier was 97 ± 1.3 nm, and the average particle diameter of the magnetically immobilized PLA 1 was 105 nm ± 1.3 nm. The enzyme activity was 1940.5 U/g. The magnetic enzyme was chemically modified with formaldehyde, dextran-aldehyde, and dextran-aldehyde-glycine. The proportions of primary amino groups in the modified magnetic immobilized enzyme PLA 1 were 0, 53.5% and 47.3%, respectively. The optimum pH of the enzyme after chemical modification was 6.5. When the system temperature was 60 °C, the magnetically immobilized PLA 1 modified with dextran-aldehyde-glycine had the optimal activity and stability. This chemically modified magnetic immobilized PLA 1 was applied to soybean oil degumming at 60 °C, 6.5 h (reaction time), and 0.10 mg/kg (enzyme dosage). The phosphorus content in the degummed oil was 9.2 mg/kg. The relative enzyme activity was 77.6% after 7 reuses which would be potentially advantageous for industrial applications.
研究领域:
生物化学
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食品科学
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PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Xuan Liu,Chunjin Huang,Dongming Lan,Weifei Wang,Yonghua Wang
Degumming is essential to eliminate undesired phospholipids from crude vegetable oils before being processed into high-quality products. The degumming efficiency is hampered by the separation of oil and water. To overcome this problem, we introduced an innovative enzymatic degumming strategy for various vegetable oils (rice bran, peanut and flaxseed oil) by combining phospholipase A1 with monoacylglycerol lipase. Compared to using phospholipase A1, the combination of the two enzymes significantly reduced phosphorus content, indicating better hydrolysis of phospholipids. Additionally, the primary hydrolysates by the combined enzymes were glycerylphosphorylcholine and glycerylphosphorylethanolamine that were more affinitive to water to improve oil-water separation and oil yield. Characteristic markers obtained from Partial Least Squares Discriminant Analysis model confirmed the advantages of combined enzymes over single enzyme. This advantage of combined enzymes was further validated in a simulated degumming process. Our findings provide a new enzymatic degumming strategy to expand the toolkit for crude oil refinery.
研究领域:
食品科学
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生物化学
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）