ANALYTICAL BIOCHEMISTRY
·
2024
·
IF 2.6
Cheng Li,Qinghui Chen,Jinqiao Rong,Houde He,Yu Lu,Yuxia Liu,Zhongfu Wang
As the main active glycoprotein of egg white, the biological functions of chicken ovomucin α- and β-subunit are closely related to the structure of glycans. However, the exact composition and structure of the subunit glycans are still unknown. We obtained highly pure chicken ovomucin α-subunit and β-subunit protein bands by the strategy combined with two-step isoelectric precipitation and SDS-PAGE gel electrophoresis. The ammonia-catalyzed one-pot procedure was then used to release and capture α-and β-subunit protein glycans with 1-phenyl- 3-Methyl-5-pyrazolone (PMP). The N / O -glycans of bis-PMP derivatives were purified and analyzed by LC-MS. More importantly, an effective dual modification was performed to accurately quantify neutral and sialylated O -glycans through methylamidation of sialic acid residues and simultaneously through carbonyl condensation reactions of reducing ends with PMP. We first showed that the α-subunit protein has only N -glycosylation modification, and the β-subunit only O -glycosylation, a total of 22 N -glycans and 20 O -glycans were identified in the α- and β-subunit, respectively. In addition, the complex N -glycan (47 %) and the sialylated O -glycan (77 %) are each major types of the above subunits. Such findings in this study provide a basis for studying the functional and biological activities of chicken ovomucin glycans.
研究领域:
生物化学
·
糖生物学
·
蛋白质组学
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DOI
PubMed
高影响因子
Foods
·
2026
·Vol. 15
·期 3
·
IF 6
Weiying Lu,Yumei Chen,Yuge Niu,Liangli (Lucy) Yu
Coffee proteins are key precursors of roasting flavor. However, heat-driven changes in the bean proteome remain underexplored. This work aimed to investigate these changes and study proteomic markers of the coffee bean. The green and roasted coffee beans were quantified for their total soluble protein and compared by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and liquid chromatography–mass spectrometry (LC–MS) proteomics. The protein profiles identified by LC–MS were processed using principal component analysis (PCA) and partial least-squares discriminant analysis (PLS–DA) modeling to identify possible roast-sensitive protein markers. The alkaline-aided aqueous extract protein concentration was reduced from 14–23 g to 3–10 g/100 g dry weight (DW). SDS–PAGE showed dominant 17–26, 34–43, and 55–72 kDa bands weakened after roasting, while high molecular peaks (>180 kDa) were present only in roasted samples. In-solution tryptic digestion yielded nine protein groups. PCA scores revealed partial separation of green and roasted groups, while PLS-DA delivered unambiguous classification (Q2> 0.90 by cross-validation). The variable importance in projection scores highlighted that structural proteins in common plant beans are markedly down-regulated after roasting, indicating heat-induced structural disruption. The identified protein groups represent candidate markers associated with severe thermal treatment and provide possible molecular targets for investigating flavor precursor development.
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PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2016
·
IF 2.857
Lingmei Li,Chengjian Wang,Shan Qiang,Jixiang Zhao,Shuang Song,Wanjun Jin,Bo Wang,Ying Zhang,Linjuan Huang,Zhongfu Wang
Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. Therefore, it can expand our knowledge about soybean protein glycosylation and provide significant structural reference for research of soybean allergens.
研究领域:
食品科学
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蛋白质组学
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分子生物学
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DOI
PubMed
高影响因子
Bioresource Technology Reports
·
2018
·
IF 0
Cheng Cai,Yu Jin,Yuxia Pang,Qia Ke,Wenhui Qiu,Xueqing Qiu,Yanlin Qin,Hongming Lou
Cellulase components in hydrolyzate were accurately traced by SDS-PAGE gel electrophoresis during the enzymatic hydrolysis of corncob residual (CCR) and delignified CCR (CCR-DL). After a quick initial adsorption on substrates, cellulase in hydrolyzate remained stable for CCR, but showed an obvious increasing trend for CCR-DL. It indicated that lignin in CCR was the main factor for the loss of cellulase in hydrolyzate. During CCR's hydrolysis, β‑glucosidase, xylanase , endoglucanase III (EGIII) and EGV were rapidly adsorbed on lignin and weren't released in subsequent, however, cellobiohydrolase I (CBHI), CBHII, EGI and EGIV were mainly adsorbed on cellulose in the early stage. With the hydrolysis of substrate, cellulase on cellulose would be released into hydrolyzate, but cellulase in hydrolyzate gradually became hydrophobic and much easier to be adsorb on lignin by the increasing hydrophobic interaction . These two factors dominated the change of cellulase concentration in hydrolyzate in the later stage of CCR's hydrolysis.
研究领域:
生物化学
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分子生物学
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植物资源利用
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酶工程
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DOI
Fermentation-Basel
·
2023
·Vol. 9
·期 8
·764
·
IF 3.7
Xinchao Yang,Linlin Niu,Chunjiang Ye,Yuanxiu Wang,Yuehui Liu,Fang Wang,Naxin Sun
D-glucaric acid is an important bio-based building block of polymers and is a high value-added chemical that can be used in a variety of applications. In the present study, theUdhtarget genes fromPseudomonas putidaandPseudomonas syringaewere used together to construct the expression vector pETDuet-2 × Udh. The transformants of BL21 (DE3) with vector pETDuet-2 × Udh were applied to produce glucaric acid from glucuronic acid. After optimizing the induction conditions, the highest Udh expression was achieved when 0.4 mmol·L−1isopropyl-β-d–thiogalactoside (IPTG) was added to the cell cultures at an OD600value of 0.6 followed by culturing at 26 °C for 6 h. The production of glucaric acid substantially reached 5.24 ± 0.015 g·L−1in fed-batch cultures in a 30 L tank. In the present study, a new system for glucaric acid production was established, which was more economic and friendly to the environment.Keywords:urinate dehydrogenases;pETDuet-2 × Udh;D-glucaric acid;glucuronic acid;SDS-PAGE
研究领域:
生物技术
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微生物学
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生化工程
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DOI
高影响因子
Journal of Chemistry
·
2020
·
IF 1.79
Di An,Liang Li
Nanofibril system constructed by protein self-assembly is widely used in the food industry because of purposive functional properties. Soy protein isolate nanofibrils (SPINs) were reported to form via heating at pH 2.0. In this research, the soy protein isolate (SPI) hydrolysate prepared by trypsin was used as a raw material for the formation of nanofibrils called soy protein isolate hydrolysate nanofibrils (SPIHNs). Microscopic images demonstrated the formation of two nanofibrils. Based on circular dichroism spectroscopy and Thioflavin T (ThT) fluorescence spectral, we concluded that β-sheet played an important role in SPIN and SPIHN’s structural composition. At the same time, the α-helix in SPI had not been destroyed, thereby favoring the formation of SPIHN. The surface hydrophobicity of SPIHN continued to increase during the heating process and reached the highest value when heating 8 h. SDS-PAGE analysis showed that peptides produced by enzyme-modified SPI affected the formation of SPIHN. These results proposed that enzymatic hydrolysis prior to acidic during fibrillation process affected the fibrillation of SPI, and the peptides formed by enzymatic hydrolysis were more efficient for the self-assembly process. This study will provide a theoretical basis for the future research of SPI nanofibril functionality.
研究领域:
食品科学
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蛋白质化学
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纳米技术
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DOI
高影响因子
Biomed Research International
·
2014
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IF 0
Jianhua Hao,Junzhong Liu,Mi Sun
A bacterial strain C5 that can produce new type of marine esterase was isolated and screened from marine sludge. According to 16S rRNA sequence analysis and physiological and biochemical experiments, the strain was identified as Bacillus subtilis. A single isozyme with a molecular weight of 86 kDa was observed by SDS-PAGE and native-PAGE. On this basis, the mechanism of esterase B1 secreted by strain C5 degrading parathion-methyl was explored, and the effects of temperature and pH on the degradation rate were investigated. From the results, p-nitrophenol was one of the degradation products of B1 degrading parathion-methyl, and the best degradation effect could be achieved at the temperature of 40°C and the neutral pH value.
研究领域:
微生物学
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环境科学
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生物技术
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DOI
PubMed
BIODEGRADATION
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2016
·Vol. 27
·期 1
·37-46
·
IF 2.208
Zhou YuYang,Huang Huanlin,Shen Dongsheng
This study evaluated substrate interactions during the aerobic biodegradation of 1, 4-dioxane and BTEX mixtures by a pure culture, Acinetobacter baumannii DD1, which is capable of utilizing 1, 4-dioxane for growth. A. baumannii DD1 could utilize BTEX as a sole carbon source, but could not utilize m -xylene and p -xylene. In binary mixtures, there was a lag of about 14 h before the degradation of BTE, and 1, 4-dioxane only started to be utilized when BTE was completely degraded by 1, 4-dioxane-grown DD1. Furthermore, the biodegradation rate of 1, 4-dioxane decreased from 73.33 to 40.74 mg/(h g dry weight) after the biodegradation of benzene. 1, 4-dioxane could not be degraded after the biodegradation of o -xylene in 80 h. DD1 could also not degrade m -xylene and p -xylene coexisting with 1, 4-dioxane. The ability of DD1 to degrade BTEX occurred in the following order: benzene > ethylbenzene > toluene > o -xylene > m -xylene = p -xylene. The biodegradation of 1, 4-dioxane was not activated in the mixture with o -xylene, primarily because of the accumulation of the specific toxic intermediate, 2, 3-dimethylphenol. The lag in BTE degradation was presumably because of the induction of enzymes necessary for BTE degradation. Additionally, SDS-PAGE analysis demonstrated that there were different proteins during the degradation of benzene and 1, 4-dioxane.
研究领域:
环境微生物学
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生物修复
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生物化学
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DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2024
·
IF 6
Yanan Wang,Xuezhi Tian,Zehua Zhang,Meiling Tian,Fusheng Zhang
The aim of this study is to enhance the efficiency of enzymes, reduce operational costs, and increase production output. Experimental research shows that ultrasound positively impacted the activity of α-amylase and glucoamylase, with the peak α-amylase activity observed at 30 W for 5 min and the highest glucoamylase activity at 60 W for 3 min. Ultrasonication extended the optimal temperature of both enzymes, but had no effect on the optimal pH. According to the kinetic experiments, ultrasonic treatment could improve the catalytic efficiency of α-amylase and glucoamylase. Furthermore, the SDS-PAGE results revealed that ultrasound treatment did not alter the molecular weights of the enzymes and their primary structures; The circular dichroism and fluorescence spectroscopy analysis indicated the secondary structural components, especially the α-sheet and the tryptophan fluorescence intensity, were notably affected by ultrasound. The changes in the secondary and tertiary structure of α-amylase and glucoamylase suggest that ultrasonic treatment may enhance their activities. Our findings provide a theoretical basis for the efficient utilization of α-amylase and glucoamylase through understanding the enzyme structure changes after ultrasound.
研究领域:
生物化学
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酶学
·
超声技术
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食品科学
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DOI
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2020
·
IF 4.389
Shouning Yang,Yanmin Zhang,Sijia Lu,Lihong Liu,Lin Yang,Yuming Guo,Shaoning Yu,Huayan Yang
One of the main diseases contributing to human death are malignant tumors. Phototherapy is a promising approach for cancer therapy, and functional nanoparticles with targeting ligands are commonly used to improve the therapeutic efficiency. However, recent studies have shown that nanoparticles in contact with a biological fluid can rapidly form a “protein corona” on their surface, which will remarkably decrease the targeting ability. Here, we describe the preparation of hybrid nanomaterials with Bi 2 S 3 nanorods as the core, and fluorescein-isothiocyanate and folic acid-modified human serum albumin (HSA-FITC-FA) as the shell. By using fluorescent binding label (FITC) and imaging techniques, we discovered the image of the cell lysosomes, indicating that the photothermal therapy agent was predominantly targeted to and accumulated in lysosomes. Combined with photothermal therapy agent (Bi 2 S 3 nanorods) and targeting ligand (FA), the obtained product shows enhanced photothermal therapy under near-infrared region laser irradiation. Additionally, SDS-PAGE shows that the modified HSA shell could remarkably reduce the reabsorption of protein corona from blood serum, minimized the adverse effect of protein corona on targetability. Taken together, the results indicate that our strategy has the potential for preparing efficient photothermal nanomaterials with image-guided subcellular organelle-targeting cancer cell ablation ability.
研究领域:
纳米技术
·
癌症治疗
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生物成像
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材料科学
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生物医学工程
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2021
·Vol. 413
·期 11
·2893-2901
·
IF 4.157
Zhang Mengmei,Wang Hao,Bhandari Ramesh,Pan Qiuhui,Sun Fenyong
Selective separation and enrichment of phosphoproteins possess the distinct clinical and biological importance in the diagnosis, treatment, and management of several fatal human diseases. In this study, a facile synthesis of titanium(IV) ion–immobilized arsenate-modified poly(glycidyl methacrylate) microparticles (denoted as Ti 4+ -arsenate-PGMA-MPs) was developed for the efficient enrichment of intact phosphoproteins found in biologically complex protein samples. By virtue of the strong interaction between the titanium ions immobilized on the surface of Ti 4+ -arsenate-PGMA-MPs and phosphate groups of phosphoproteins, Ti 4+ -arsenate-PGMA-MPs had a high saturated adsorption capacity for phosphoproteins (901 mg/g for β-casein), which was much higher than that of non-phosphoproteins (73.5 mg/g for BSA). Ti 4+ -arsenate-PGMA-MPs were characterized by SEM, TEM, and FT-IR, and the average particle diameter was about 2.5 μm with good dispersibility. Besides, the application of Ti 4+ -arsenate-PGMA-MPs in real biological samples was investigated by SDS-PAGE analysis, and the results showed that Ti 4+ -arsenate-PGMA-MPs were able to enrich phosphoproteins efficiently. Graphical abstract
研究领域:
生物化学
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蛋白质组学
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材料科学
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DOI
PubMed
高影响因子
MOLECULES
·
2024
·Vol. 29
·期 7
·1656
·
IF 4.2
Qian Zhang,Linlin Hu,Jianyu Yang,Pengfei Guo,Jinhong Wang,Weifen Zhang
Immobilized metal ion affinity chromatography (IMAC) adsorbents generally have excellent affinity for histidine-rich proteins. However, the leaching of metal ions from the adsorbent usually affects its adsorption performance, which greatly affects the reusable performance of the adsorbent, resulting in many limitations in practical applications. Herein, a novel IMAC adsorbent, i.e., Cu(II)-loaded polydopamine-coated urchin-like titanate microspheres (Cu-PDA-UTMS), was prepared via metal coordination to make Cu ions uniformly decorate polydopamine-coated titanate microspheres. The as-synthesized microspheres exhibit an urchin-like structure, providing more binding sites for hemoglobin. Cu-PDA-UTMS exhibit favorable selectivity for hemoglobin adsorption and have a desirable adsorption capacity towards hemoglobin up to 2704.6 mg g−1. Using 0.1% CTAB as eluent, the adsorbed hemoglobin was easily eluted with a recovery rate of 86.8%. In addition, Cu-PDA-UTMS shows good reusability up to six cycles. In the end, the adsorption properties by Cu-PDA-UTMS towards hemoglobin from human blood samples were analyzed by SDS-PAGE. The results showed that Cu-PDA-UTMS are a high-performance IMAC adsorbent for hemoglobin separation, which provides a new method for the effective separation and purification of hemoglobin from complex biological samples.
研究领域:
生物化学
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材料科学
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生物技术
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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）