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CARBOHYDRATE POLYMERS · 2025 · IF 12.5

Development of derivatization-enhanced EIEIO-driven glycosidic linkage sequencing (DEED-GL-Seq) strategy and its application to glycogen-sourced oligosaccharides profiling

Xin Zheng,Yufang Ma,Xinge Cui,Yi Wang,Jingjing Jiang,Ting Liu,Zhijun Zhang,Mingsheng Ma,Xiaohong Han,Cai Tie

The structural complexity arising from diverse glycosidic linkages in oligosaccharides hinders the elucidation of their biological functions. Existing analytical methods often lack the necessary feasibility and accessibility for comprehensive linkage analysis. To address this, we developed Derivatization-Enhanced EIEIO-Driven Glycosidic Linkage Sequencing (DEED-GL-Seq), a novel strategy leveraging differential electron density modulation around glycosidic bonds upon N 2 ,N 2 ,N 4 ,N 4 -tetraethyl-6-hydrazineyl-1,3,5-triazine-2,4-diamine (T3) derivatization. This method integrates EIEIO MS 2 to identify glycosidic bond-specific diagnostic fragments for linkage determination. Validation confirmed DEED-GL-Seq's specificity, sensitivity, reliability, and standard-independence. Coupling with fine-tuned HILIC separation further enhanced its analytical power for complex matrices. We applied DEED-GL-Seq to profile oligosaccharides derived from partially hydrolyzed glycogen, validating our results against reference standards. By analyzing the abundance ratios of specific trisaccharides, we predicted the branching degrees of glycogen and amylopectin, corroborating existing literature. Notably, DEED-GL-Seq identified established (Glc₄) and novel (HEX-1, HEX-2, HEPTA-1, HEPTA-2) potential glycosidic biomarkers for GSD-II in urine, with the novel oligosaccharides showing superior diagnostic performance. The unique structure of HEX-2 suggests distinct biosynthetic pathways in GSD-II pathophysiology. DEED-GL-Seq represents a significant advancement in glycomics, offering a powerful tool for comprehensive oligosaccharide profiling and laying a foundation for in-depth functional investigations. This work presents a novel application paradigm for EIEIO technology.

CAS / MDL: 34612-38-9 35997-20-7
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES · 2026 · IF 8.7

Bioinformatics-assisted mining and engineering of α-amylase for efficient hydrolysis of raw corn and microalgal starch

Xueting Zhang,Tong Ye,Yanli Zhang,Li Huang,Yimeng Song,Xucheng Zhang,Yinliang Zhang,Yazhong Xiao,Wei Fang

Raw starch-degrading α-amylases play a key role in the cold hydrolysis of starch granules. Using bioinformatic approaches, one α-amylase-encoding gene, AmySy , was identified in the database. AmySy exhibited high activity toward raw corn and microalgal starch, with specific activities of 7570 ± 234 and 2564 ± 33 U/mg, respectively. Through a combination of computational design strategies, the variant AmySy:G207H was obtained, which exhibited enhanced catalytic performance. The half-life of AmySy:G207H at 35 °C was extended to 113 h, representing a 3.0-fold increase over AmySy. Using 30% raw corn flour as the substrate, the hydrolysis rate of AmySy:G207H reached 34.7% after a 5-h reaction. A chemo-enzymatic saccharification process for microalgae was developed using AmySy:G207H. After a three-hour reaction, the reducing sugar yield reached 367.5 mg/g microalgal biomass, which was 2.5 times that obtained through chemical hydrolysis alone. Owing to its ability to hydrolyze raw corn and microalgal starch, as well as its excellent thermostability, AmySy:G207H can be considered an efficient enzyme for the conversion of raw starch into sugar.

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JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS · 2023 · IF 3.4

Characterization and comparison of polysaccharides from Achyranthes bidentata, Cyathula officinalis and Achyranthes aspera by saccharides mapping

HuaYang Si,Yimeng Chen,Jie Yang,Xiaodong Wen

Although the roots of Achyranthes bidentata (AB), Cyathula officinalis (CO) and Achyranthes aspera (AA) are different drugs, they are always confused in clinical practice due to their similar chemical components and functions. As polysaccharides are abundant in these drugs, a systematic comparison of polysaccharides from AB, CO and AA is not only necessary to understand their similar but not identical functions, but also helpful for the quality control of them. In this study, polysaccharides from 22 batches of AB, CO and AA were compared with monosaccharide composition , molecular weight distribution and saccharide mapping. Polysaccharides of AB, CO and AA had similar monosaccharide compositions but their relative contents of fructose, glucuronic acid , galacturonic acid and glucose were significant different, and could be used as key markers to distinguish them. Results from molecular weight distribution and saccharide mapping showed polysaccharides from AB, CO and AA were mainly composed of fructans with β-2,1 and β-2, 6- D -fructosidic linkages, but their degree of polymerization were different. Meanwhile, pectins were also contained in these three drugs. AB is partial to immunomodulation while CO is partial to removing blood stasis . Fructans and pectins are the similar bioactive substance basis of AB, CO and AA whereas their structural difference might be contributed to the efficacy differentia of these three drugs. This study provides a better understanding on the profiles of polysaccharides from AB, CO and AA, further guiding their clinical usage and facilitating their quality control.

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Journal of Ginseng Research · 2021 · IF 6.06

Evaluation of storage period of fresh ginseng for quality improvement of dried and red processed varieties

Na Zhang,Xin Huang,Yun-Long Guo,Hao Yue,Chang-Bao Chen,Shu-Ying Liu

Background Dried and red ginseng are well-known types of processed ginseng and are widely used as healthy food. The dried and red ginseng quality may vary with the storage period of raw ginseng. Therefore, herein, the effect of the storage period of fresh ginseng on processed ginseng quality was evaluated through multicomponent quantification with statistical analysis. Methods A method based on ultrahigh performance liquid chromatography coupled to triple quadrupole mass spectrometry in multiple-reaction monitoring mode (UPLC-MRM-MS) was developed for quantitation of ginsenosides and oligosaccharides in dried and red ginseng. Principal component analysis and partial least squares discriminant analysis were conducted to evaluate the dynamic distributions of ginsenosides and oligosaccharides after different storage periods. Results Eighteen PPD, PPT and OLE ginsenosides and nine reducing and nonreducing oligosaccharides were identified and quantified. With storage period extension, the ginsenoside content in the processed ginseng increased slightly in the first 2 weeks and decreased gradually in the following 9 weeks. The content of reducing oligosaccharides decreased continuously as storage time extending, while that of the nonreducing oligosaccharides increased. Chemical conversions occurred during storage, based on which potential chemical markers for the storage period evaluation of fresh ginseng were screened. Conclusion According to ginsenoside and oligosaccharide distributions, it was found that the optimal storage period was 2 weeks and that the storage period of fresh ginseng should not exceed 4 weeks at 0 °C. This study provides deep insights into the quality control of processed ginseng and comprehensive factors for storage of raw ginseng.

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JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE · 2020 ·Vol. 100 ·期 7 ·3062-3070 · IF 2.614

Profiling of carbohydrates in commercial beers and their influence on beer quality

Miaomiao Li,Jinhua Du,Kaili Zhang

BACKGROUND The carbohydrates in beer play an important role as they are essential for fermentation. Any change in their composition may influence the sensory characteristics of the beer and so their determination is of great interest. This study compares the carbohydrates in three types of commercial beer – barley malt beer, wheat beer, and barley malt beer with adjuncts – and examines their influence on beer quality, which is important for selecting raw ingredients and production conditions, and for quality control. RESULTS Among the oligosaccharides in three types of beer, raffinose was the most, followed by maltotetraose, maltotriose and maltose. Monosaccharides were only present in small amounts. Dextrin, oligosaccharides with 2–6 polymerization degree and non-starch polysaccharides (NSP) make up 15.90–34.83%, 17.59–38.63%, and 2.33–7.47% of the total carbohydrates in beer, respectively. The dextrin content and NSP content were significantly ( P < 0.05) different in wheat beer and barley malt beer, and their content was significantly ( P < 0.01) correlated with the content of extracts in beer. Non-starch polysaccharide, dextrin, trisaccharide, and tetrasaccharide content significantly ( P < 0.05) correlated with beer viscosity. These beer samples could be categorized clearly into three groups by principal component analysis. CONCLUSION The oligosaccharides in beer reflect yeast utilization, depending on the type of beer. Dextrin, oligosaccharides with 2–4 polymerization, and NSP, were major carbohydrates in beer. Their composition and concentration influenced its characteristics and quality, and played an important role in the discrimination of different beer types. © 2020 Society of Chemical Industry

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ANALYTICA CHIMICA ACTA · 2019 · IF 5.256

Copper-based two-dimensional metal-organic framework nanosheets as horseradish peroxidase mimics for glucose fluorescence sensing

Mei-Yuan Shi,Ming Xu,Zhi-Yuan Gu

Ultrathin two-dimensional (2D) metal-organic framework (MOF) nanosheets have attracted numerous attention due to their unparalleled advantages such as unique structures, large surface areas, good stability and high catalytic efficiency . In this paper, 2D Cu(bpy) 2 (OTf) 2 nanosheets (Cu-MOF, bpy  = 4,4-bipyridine, OTf = trifluoromethanesulfonate) are first utilized to achieve the fluorescent detection for H 2 O 2 and glucose with their intrinsic peroxidase-like activity. The fluorescent biosensor Cu-MOF nanosheets were achieved by the efficient catalysis of non-fluorescent thiamine (TH) to strong fluorescent thiochrome in the presence of H 2 O 2 . With the combination of glucose oxidase , highly selective and sensitive fluorescent detection for glucose could be established with a detection limit of 0.41 μM and two separate linear ranges of 10–100 μM and 100–1000 μM, respectively. Furthermore, the developed method has been applied to human serum for the quantitative determination of glucose as a clinical diagnosis indicator of diabetes mellitus.

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ACS Catalysis · 2018 · IF 11.384

Stoichiometric Conversion of Cellulosic Biomass by in Vitro Synthetic Enzymatic Biosystems for Biomanufacturing

Dongdong Meng,Xinlei Wei,Yi-Heng P. Job Zhang,Zhiguang Zhu,Chun You,Yanhe Ma

Cellulosic biomass is the earth’s most abundant renewable resource, which is considered to be a promising feedstock for manufacturing biofuels and biochemicals. In this study, stoichiometric enzymatic phosphorolysis of cellulosic biomass for manufacturing biochemicals or biofuels by in vitro synthetic enzymatic biosystems was designed. Three cascade phosphorolytic enzymes, cellodextrin phosphorylase, cellobiose phosphorylase, and polyphosphate-dependent glucokinase, were used for the biotransformation of cellodextrins to high-energy phosphorylated sugars (that is, glucose 1-phosphate and glucose 6-phosphate). A series of downstream exergonic reactions then converted these high-energy phosphorylated sugars into myo-inositol, resulting in a near-stoichiometric conversion of cellodextrins with a high product yield of 98% (w/w). Moreover, this enzymatic biosystem can even work for the acid-treated biomass hydrolysate containing microorganism-toxic compounds. The construction of this in vitro synthetic enzymatic biosystem provided an alternative method for the utilization of cellulosic biomass rather than cellulolytic enzyme hydrolysis to fermentative monomeric sugars followed by microorganism fermentation, showing potentials in the production of biocommodities such as hydrogen, rare sugars, and electricity from cellulosic biomass.

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APPLIED AND ENVIRONMENTAL MICROBIOLOGY · 2015 ·Vol. 81 ·期 6 · IF 3.668

AmyM, a Novel Maltohexaose-Forming α-Amylase from Corallococcus sp. Strain EGB

Zhoukun Li,Jiale Wu,Biying Zhang,Fei Wang,Xianfeng Ye,Yan Huang,Qiang Huang,Zhongli Cui

A novel α-amylase, AmyM, was purified from the culture supernatant of Corallococcus sp. strain EGB. AmyM is a maltohexaose-forming exoamylase with an apparent molecular mass of 43 kDa. Based on the results of matrix-assisted laser desorption ionization–time of flight mass spectrometry and peptide mass fingerprinting of AmyM and by comparison to the genome sequence of Corallococcus coralloides DSM 2259, the AmyM gene was identified and cloned into Escherichia coli. amyM encodes a secretory amylase with a predicted signal peptide of 23 amino acid residues, which showed no significant identity with known and functionally verified amylases. amyM was expressed in E. coli BL21(DE3) cells with a hexahistidine tag. The signal peptide efficiently induced the secretion of mature AmyM in E. coli. Recombinant AmyM (rAmyM) was purified by Ni-nitrilotriacetic acid (NTA) affinity chromatography, with a specific activity of up to 14,000 U/mg. rAmyM was optimally active at 50°C in Tris-HCl buffer (50 mM; pH 7.0) and stable at temperatures of <50°C. rAmyM was stable over a wide range of pH values (from pH 5.0 to 10.0) and highly tolerant to high concentrations of salts, detergents, and various organic solvents. Its activity toward starch was independent of calcium ions. The Km and V max of recombinant AmyM for soluble starch were 6.61 mg ml−1 and 44,301.5 μmol min−1 mg−1, respectively. End product analysis showed that maltohexaose accounted for 59.4% of the maltooligosaccharides produced. These characteristics indicate that AmyM has great potential in industrial applications.

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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES · 2024 · IF 7.7

Optimization and analysis of dual-enzymatic synthesis for the production of linear glucan

Yue Guo,Shenglin Sun,Mingfei Gu,Liyue Zhang,Li Cheng,Zhaofeng Li,Caiming Li,Xiaofeng Ban,Yan Hong,Zhengbiao Gu

Linear α-glucan (LG), a linear polymer linked by α-1,4 bonds, has received increasing attention for its potential applications in synthetic polymer production. Notably, the functionality of LG is strongly influenced by its degree of polymerization (DP). In this study, SP and GP were successfully constructed and expressed. The reaction of enzymatic co -polymerization into LG was investigated. The preferred reaction was carried out at 37 °C and pH 7.4 for 72 h, with a maximum conversion rate of 25 %. Afterwards, two approaches were used to modulate the molecular structures of LGs. Firstly, LGs with distinct molecular weights ranging from 1062.33 ± 16.04 g/mol to 5679 ± 80.29 g/mol were obtained by adjusting the substrate/primer ratio during the LG synthesis process. Secondly, two distinct products could be produced by altering the enzyme addition method: short-chain LG with a DP < 10 (64.34 ± 0.54 %) or long-chain LG with a DP > 45 (45.57 ± 2.18 %). Additionally, theoretical synthesis model was constructed which subdivided the reaction into three stages to evaluate this dual-enzyme cooperative system. These findings have significant implications in promoting the application of LG in the fields of biomedicine and material science.

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Journal of Advanced Research · 2025 · IF 13

Saccharide mapping apparatus for real-time PAGE detection of polysaccharides

Baojie Zhu,Jing Zhao,Shaoping Li

Introduction As a key technique in saccharide mapping, polysaccharides analysis using carbohydrate gel electrophoresis (PACE) analysis platform has always been self-assembled and built in various laboratories independently, but there is no unified commercial analysis instrument platform so far. Objective This research aimed to establish an integrated commercially viable PACE analysis platform to standardize the application of PACE in the field of saccharides analysis and the quality control of polysaccharides. Methods We established an online detection platform of saccharide-mapping apparatus (SMA) and took advantage of it to characterize the oligosaccharides for quality control of polysaccharides in herbal medicine. The polysaccharides from Gastrodia elata Bl. were finally selected to verify the potential of SMA including precision, repeatability, stability, and sensitivity. Results The online detection platform of SMA was more integrated and commercially viable, which can investigate oligosaccharides from polysaccharides after hydrolysis more quickly and accurately for quality control. Consequently, it could enable real-time monitoring of oligosaccharide migration, facilitate the determination of the optimal endpoint of polyacrylamide gel electrophoresis (PAGE), and improve the sensitivity of PACE. In addition, the refrigeration device configured in SMA can solve the problem of temperature fluctuation caused by joule heat during gel electrophoresis, which greatly enhances the separation resolution. Finally, the proposed platform was applied to real samples of polysaccharides from Gastrodia elata Bl., and the results successfully distinguished the polysaccharides from Gastrodia elata Bl. with different cultivars. Conclusion The developed online detection platform of SMA offers a rapid and convenient approach for quality control of polysaccharides in Chinese medicines and can be applied for future advancements toward commercialization.

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