JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2023
·
IF 6.1
Simin Liu,Jiamei Liu,Linguo Zhao,Jianjun Pei
Glycosylation, one of the most common and significant modifications in nature, has prompted the development of a cellobiose phosphorolysis route for glycosylation in vivo. However, the process of glycosylation is hampered by the notably low conversion rate of cellobiose. In this work, regulation of the carbon source supply by changing the ratio of glucose to cellobiose improved the conversion rate of cellobiose, resulting in enhancing the efficiency of glycosylation and the production of vitexin. Moreover, three genes (pgm, agp, and ushA) involved in the degradation of UDP-glucose were knocked out to relieve the degradation and diversion of the cellobiose phosphorolysis route. Finally, through the optimization of conversion conditions, we observed a continuous enhancement in cellobiose conversion rate and vitexin production in BL21ΔushAΔagp-TcCGT-CepA, corresponding to an increased concentration of added glucose. The maximum production of vitexin reached 2228 mg/L with the addition of 2 g/L cellobiose and 6 g/L glucose, which was 312% of that in BL21-TcCGT-CepA with the addition of 2 g/L cellobiose. The conversion rate of cellobiose in BL21ΔushAΔagp-TcCGT-CepA reached 88%, which was the highest conversion rate of cellobiose to date. Therefore, this study presents a cost-effective and efficient method to enhance the conversion rate of cellobiose during the glycosylation process.
研究领域:
生物化学
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代谢工程
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DOI
PubMed
高影响因子
ENVIRONMENTAL MICROBIOLOGY
·
2021
·Vol. 23
·期 9
·5147-5163
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IF 5.491
Can Zhang,Meng Cai,Shanshan Chen,Fan Zhang,Tongshan Cui,Zhaolin Xue,Weizhen Wang,Borui Zhang,Xili Liu
Summary Asparagine (Asn, N )-linked glycosylation within N glyco -X-S/T; X ≠ P motif is a ubiquitously distributed post-translational modification that participates in diverse cellular processes. In this work, N -glycosylation inhibitor was shown to prevent Phytophthora sojae growth, suggesting that N -glycosylation is necessary for oomycete development. We conducted a glycoproteomic analysis of P. sojae to identify and map N -glycosylated proteins and to quantify differentially expressed glycoproteins associated with mycelia, asexual cyst, and sexual oospore developmental stages. A total of 355 N -glycosylated proteins was found, containing 496 glycosites, potentially involved in glycan degradation, carbon metabolism, glycolysis, or other metabolic pathways. Through PNGase F deglycosylation assays and site-directed mutagenesis of a GPI transamidase protein (GPI16) upregulated in cysts and a heat shock protein 70 (HSP70) upregulated in oospores, we demonstrated that both proteins were N -glycosylated and that the N glyco -N motif is a target site for asparagine – oligosaccharide linkage. Glycosite mutations of Asn 94 N glyco -X-S/T in the GPI16 led to impaired cyst germination and pathogenicity, while mutation of the previously unknown Asn 270 N glyco -N motif in HSP70 led to decreased oospore production. In addition to providing a map of the oomycete N -glycoproteome, this work confirms that P. sojae has evolved multiple N -glycosylation motifs essential for growth.
研究领域:
分子生物学
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植物病理学
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微生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Tong Zhao,Sining Liu,Yutong Shuai,Xinyi Zhang,Min Chen,Sijie Pei,Yuxi Duan,Shukai Wang,Yu Lu,Zhongfu Wang,Guiping Gong,Linjuan Huang
Pectin is a structurally complex heteropolysaccharide that affects intestinal microorganisms and mucin O -glycans. The present study employed an in vitro model to investigate dynamic changes in microbiota during pectin fermentation. Residual pectin fragments arising from its fermentation were applied to HT29-MTX cells to study the effect of pectin structure on mucin O -glycosylation. Prevotella , Bacteroides , and Parabacteroides were found to preferentially degrade galactose, arabinose, and on the rhamnogalacturonan RG-I side chain region and methyl esterification groups of pectin. Bifidobacterium , Enterococcus , Megamonas , and Dorea metabolized the galacturonic HG region on pectin to produce butyrate. All pectin fragments were found to up-regulate total O -glycans (1.55–2.73 fold) and neutral O -glycans (1.11–1.49 fold) on HT29-MTX mucins. The large HG fragment (81.04 kDa) increased significantly the amount of non-fucosylated glycans (by 2.46-fold); whereas the small HG fragment (16.02 kDa) promoted fucosylated (by 9.25 fold), and especially di-fucosylated O -glycans. Collectively, these results demonstrate that gut microorganisms degrade pectin fragments in the following order of utilization: RG-I, RG-II, and HG. The small fragment of HG improves the expression of fucosylated O -glycans in HT29-MTX cells, mainly owing to an increase in di-fucosylated O -glycans.
研究领域:
微生物学
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胃肠病学
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细胞生物学
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营养科学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Xiang Ye,Shengqi Jiang,Wei Niu,Rong Bai,Chunjie Yang,Siying Wang,Ziwei Li,Linlu Zhang,Huiyu Han,Jing Xi,Wu Ding
The influence of electron beam irradiation (EBI) treatment on the modification of gelatin-galactose glycosylation was thoroughly examined. The results of the degree of grafting and browning revealed that EBI triggered the glycosylation reaction of gelatin. The degree of glycosylation exhibited a gradual increase with the rising irradiation dose, reaching a maximum of 25 kGy. Moreover, the irradiation process opened up gelatin's internal structure, exposing its hydrophobic groups. This exposure led to an enhancement in sample surface hydrophobicity. The fluorescence intensity at the maximum emission wavelength of the fluorescence spectra decreased; Fourier infrared spectroscopy demonstrated a new absorption peak at 1074 cm −1 for the glycosylation product. These findings substantiate that gelatin formed a new product through covalent bonding with galactose. Glycosylation boosted the emulsification stability of gelatin from 1.92 min to 10.42 min and improved its emulsification and rheological properties. These outcomes affirm that EBI can effectively induce the glycosylation reaction of gelatin, thereby enhancing its functional properties. In addition, EBI has the potential to supplant the conventional heating glycosylation method. This study lays a solid theoretical foundation for the future application of glycosylation and gelatin.
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
JOURNAL OF CARBOHYDRATE CHEMISTRY
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2016
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IF 0.738
Faisal Nureldin Awad,Anna Kulinich,Ming Jun Yao,Xu Chu Duan,Zhi Peng Cai,Bin Gu,Li Liu,Josef Voglmeir
GRAPHICAL
研究领域:
生物化学
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酶学
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DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2023
·
IF 8.2
Jing-Jing Liu,Zhan-Yi Wang,Bin-Bin Jiang,Shu-Qin Gao,Ying-Wu Lin
Nonenzymatic glycation and the subsequent accumulation of advanced glycation end-products (AGEs) in proteins are factors underlying long-term pathogenesis in diabetes. The study of protein glycation is crucial for elucidating their relationship with diabetes mellitus and related disorders. This study explores the interaction between d -ribose and human myoglobin (HMb), as well as the protective effect of thymoquinone (TQ) on glycation. A time-dependent in-vitro glycation study was performed to investigate the mechanism of d -ribose-induced structural interference of HMb in the absence and presence of TQ. Spectroscopic and proteomic analysis indicated that the presence of TQ significantly reduced the total amount of AGEs while maintaining structural characteristics of HMb. 14 glycated sites on HMb were further identified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) after incubation with d -ribose for 12 h, predominantly interacting with lysine residues. TQ was found to disrupt this interaction, reducing the glycated sites from 14 to 12 sites and the percentage of glycated peptides from 26.50 % to 12.97 %. Additionally, there was a significant decrease in the degree of glycation at the same sites. In summary, our findings suggest that TQ has the potential to act as an anti-glycation agent and provide a comprehensive understanding underlying the inhibition mechanism of glycation.
研究领域:
生物化学
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医学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2023
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IF 7.4
Jiajia Li,Shuang Yue,Ziyuan Gao,Wenhua Hu,Zhaoliang Liu,Guoqiang Xu,Zhen Wu,Xumin Zhang,Guolin Zhang,Fuliang Qian,Junhong Jiang,Shuang Yang
Ribonuclease (RNA) modifications can alter cellular function and lead to differential immune responses by acting as discriminators between RNAs from different phyla. RNA glycosylation has recently been observed at the cell surface, and its dysregulation in disease may change RNA functions. However, determining which RNA substrates can be glycosylated remains to be explored. Here, we develop a solid-phase chemoenzymatic method (SPCgRNA) for targeting glycosylated RNAs, by which glycosylated RNA substrates can be specifically recognized. We found the differential N-glycosylation of small RNAs in hTERT-HPNE and MIA PaCa-2 cancer cells using SPCgRNA. RNA-Seq showed that the changes in glyco-miRNAs prepared from SPCgRNA were consistent with those of traditional methods. The KEGG signaling pathway analysis revealed that differential miRNA glycosylation can affect tumor cell proliferation and survival. Further studies found that NGI-1 significantly inhibited the proliferation, migration, and circulation of MIA PaCa-2 and promoted cell apoptosis. In addition, β-1,4-galactosyltransferase 1 (B4GALT1) not only affected the expression level of glycosylated miRNAs hsa-miR-21-5p but also promoted cell apoptosis and inhibited the cell cycle possibly through the p53 signaling pathway, while B4GALT1 and p53 were also affected following the hsa-miR-21-5p increase. These results suggest that B4GALT1 may catalyze miRNAs glycosylation, which further promotes cancer cell progression.
研究领域:
分子生物学
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生物化学
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肿瘤学
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DOI
PubMed
高影响因子
DRUG DEVELOPMENT RESEARCH
·
2025
·Vol. 86
·期 1
·e70051
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IF 4.2
Qiwen Shi,Tao Zhou,Wenjun Hou,Yuqi Zhou,Shufen Deng,Ying Song
Diabetes nephropathy (DN) is a severe diabetic chronic microvascular complication and the major cause of end-stage renal disease (ESRD). Our study aimed to investigate the effects of isoliquiritigenin (ISL) a natural flavonoid compound on DN and to explore the underlying mechanisms. The db/db mice were received intragastric treatments of ISL (5, 10, or 20 mg/kg), vehicle or positive drug metformin (300 mg/kg) once a day for 12 weeks, and the db/m mice treated with vehicle were used as controls. ISL significantly ameliorated pathological changes and functional injury in the kidneys of db/db mice in a dose-dependent manner. The administration of 20 mg/kg ISL reduced the levels of serum creatinine (Scr; 49.0 ± 1.7 vs. 56.9 ± 2.9 μmol/L; p < 0.01), blood urine nitrogen (BUN; 9.6 ± 1.3 vs. 12.0 ± 1.1 mmol/L; p < 0.05), albumin to creatinine ratio (ACR; 1925.8 ± 798.1 vs. 4269.4 ± 925.6 μg/mg; p < 0.01) and urinary albumin (276.2 ± 39.9 vs. 576.9 ± 108.9 μg; p < 0.05). Further study identified advanced glycation end product (AGE)–receptor for AGE (RAGE) axis as a target of ISL. ISL (20 mg/kg) lowered renal AGE level (2.5 ± 0.5 vs. 3.8 ± 0.6 μg/mg; p < 0.01) and RAGE expression, leading to improvements in renal fibrosis, oxidative stress, and inflammation. To sum up, our study demonstrated that ISL displayed preventive effects on the experimental model of DN through suppressing AGE–RAGE pathway, and provided some insights into the application of ISL in DN treatment.
研究领域:
药理学
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内分泌学
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肾病学
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DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR LIQUIDS
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2024
·
IF 5.3
Yanqing Zhang,Lixia Yuan,Qiulu Wang,Xuejie Zhang,Yiyun Yang,Min Liu
The accumulation of advanced glycation end products (AGEs) generated from glycation reaction accelerates the development and deterioration of some diseases. Herein, the AGEs model of fructose-induced glycation of human serum albumin (HSA) was constructed and characterized. Both caffeic acid (CA) and dihydrocaffeic acid (DHCA) can inhibit the formation of AGEs, and the inhibitory effect of CA was more pronounced, as confirmed by fluorescence spectroscopy, gel electrophoresis, scanning electron microscopy, and carbonyl and thiol contents analysis. The binding of CA and DHCA to HSA was investigated by fluorescence spectroscopy and molecular docking. The primary inhibitory mechanism was the occupation of the glycation site LYS199 of HSA, and the more significant inhibitory effect of CA was related to its stronger binding affinity. Furthermore, the conformational changes caused by glycation and CA/DHCA binding were monitored using fluorescence, circular dichroism, and dynamic light scattering analysis. The results would help to elucidate the inhibitory mechanism of CA and DHCA on AGEs formation based on their different structures and inhibitory effects, and provide guidance for choosing natural inhibitors for diabetes.
研究领域:
生物化学
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药理学
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分子生物学
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高影响因子
Molecular Catalysis
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2022
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IF 5.089
Ruizhi Han,Baocheng Chai,Yulin Jiang,Jie Ni,Ye Ni
Poor water solubility limits the application of genistein in food and pharmaceutical fields. Glycosylation can greatly improve the water solubility of genistein by attaching long chain sugars. Here, glycosylation of sophoricoside (genistein monoglucoside) by cyclodextrin glycosyltransferase from Paenibacillus macerans ( Pm CGTase) was investigated. Based on molecular docking and hydrolytic activity analysis, saturation mutagenesis was conducted at residues Y195 and S77 of Pm CGTase. Variants S77N, Y195S, Y195I, S77N/Y195S, and S77N/Y195I showed enhanced product specificity toward long-chain glycosylated sophoricosides (LCGS). In particular, S77N/Y195I displayed 119% increased LCGS product specificity and 20.6% higher sophoricoside conversion than the wild type. Increased comprehensive activities of Y195S and Y195I contributed to enhanced conversion, and decreased hydrolytic activity of S77N promoted LCGS accumulation. Kinetics results revealed improved catalytic efficiency and substrate affinity of variants toward both the glycosyl donor and acceptor. Furthermore, MD simulation elucidates that the decreased substrate binding energy of variants could lead to improved kinetics, and increased hydrogen binding between residues at -6/-7 subsites and the glycosyl donor is primarily responsible for enhanced LCGS specificity. An “Increased Income-Reduced Expenditure” mechanism was proposed, explaining that the enhanced LCGS product specificity of Pm CGTase is closely related to the increased hydrogen bond interactions at -6/-7 subsites and decreased hydrolytic activity.
研究领域:
生物化学
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酶工程
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药学
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食品科学
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DOI
高影响因子
FOOD HYDROCOLLOIDS
·
2023
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IF 10.7
Hulin Geng,Wanyi Sun,Shengnan Zhan,Ru Jia,Qiaoming Lou,Tao Huang
In this study, the influence of glycosylation with κ-Carrageenan (κC), high methoxyl pectin (HMP), d -Galactose (Gal) on the gelling, rheological properties and mechanisms of fish gelatin (FG) were comparatively investigated. Results showed that κC was easiest to glycosylated with FG in comparison to HMP and Gal, also had the most significant improvement on the gelling properties and viscosity of FG. HMP was most difficult to glycosylated FG, but still had an excellent enhancement on viscosity. The results of intrinsic fluorescence, FTIR, X-Ray and SEM indicated that proper glycosylation changed the conformation structure and improved the contents of hydrogen bonds and α-helix, stabilizing the gels network. LC-MS/MS results showed that the main glycosylation sites were lysine and arginine amino groups, which was regarded as N-terminal glycosylation. In conclusion, proper N- glycosylation could significantly improve the gelling and rheological properties of FG. This study may provide a new guideline for modification FG to expand its application.
研究领域:
食品科学
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生物技术
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材料科学
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DOI
高影响因子
ACS Omega
·
2022
·
IF 4.132
Moaz M. Abdou,Paul M. O’Neill,Eric Amigues,Magdalini Matziari
A novel TEA-catalyzed sugar-esterification of phosphinic acids was used as a general and efficient approach for the synthesis of a variety of phosphinates without any transition metal. The high efficiency of the current methodology and a convenient experimental procedure compensate for the moderate yields obtained. Another advantage is that the reaction tolerates different substituents attached to the phosphinic acids and the sugar moieties alongside the ease of isolation of the product.
研究领域:
有机化学
·
合成化学
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DOI
PubMed
高影响因子