INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2026
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IF 8.7
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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Microbiology Spectrum
·
2021
·Vol. 9
·期 3
·
IF 7.171
Alia Rizvi Syeda Kazim,Yuting Jiang,Shengnan Li,Xiaoxiao He
α-Glucan is a major cell wall component and a virulence and adhesion factor for fungal cells. However, the biosynthetic pathway of α-glucan was still unclear. α-Glucan was shown to be composed mainly of 1,3-glycosidically linked glucose, with trace amounts of 1,4-glycosidically linked glucose. Besides the α-glucan synthetases, amylase-like proteins were also important for α-glucan synthesis. In our previous work, we showed that Aspergillus nidulans AmyG was an intracellular protein and was crucial for the proper formation of α-glucan. In the present study, we expressed and purified AmyG in an Escherichia coli system. Enzymatic characterization found that AmyG mainly functioned as an α-amylase that degraded starch into maltose. AmyG also showed weak glucanotransferase activity. Most intriguingly, supplementation with maltose in shaken liquid medium could restore the α-glucan content and the phenotypic defect of a ΔamyG strain. These data suggested that AmyG functions mainly as an intracellular α-amylase to provide maltose during α-glucan synthesis in A. nidulans.IMPORTANCE Short α-1,4-glucan was suggested as the primer structure for α-glucan synthesis. However, the exact structure and its source remain elusive. AmyG was essential to promote α-glucan synthesis and had a major impact on the structure of α-glucan in the cell wall. Data presented here revealed that AmyG belongs to the GH13_5 family and showed strong amylase function, digesting starch into maltose. Supplementation with maltose efficiently rescued the phenotypic defect and α-glucan deficiency in an ΔamyG strain but not in an ΔagsB strain. These results provide the first piece of evidence for the primer structure of α-glucan in fungal cells, although it might be specific to A. nidulans.
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Molecular Omics
·
2021
·Vol. 17
·期 4
·565-571
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IF 3.743
Lujie Yang,Quanqing Zhang,Ling Lin,Ying Xu,Yuanyu Huang,Zuojian Hu,Ke Wang,Cuiping Zhang,Pengyuan Yang,Hongxiu Yu
Glycosylation is a widely occurring posttranslational modification. Here, we applied a quick, convenient and high-throughput strategy (lectin array) to investigate the variation in glycans on different macrophage subtypes derived from THP-1 and RAW264.7 cells. For THP-1 cells, there were more significant differences in the glycan on M2 macrophages compared to the other two subtypes. In contrast, M1 macrophages exhibited more significant glycan remodeling than the other subtypes for the RAW264.7 cell line. The response of the lectins which recogonize the N-glycan and α2,6 sialic acid was higher during polarization into anti-inflammatory phase (THP-1 derived M2 subtypes), and lower in pro-inflammatory phase (RAW264.7 M1 subtypes). The regulation of several α2,6 sialyltransferase genes was coincident with the regulation of the α2,6 sialic acid on the two cell lines. The lectin response and glycosyltranferase gene expression confirmed that α2,6 sialic acid showed higher expression in the anti-inflammatory phase. This indicated that α2,6 sialic acid was a potential indicator for the anti-inflammatory response.
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Journal of Ginseng Research
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2021
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IF 6.06
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
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2020
·Vol. 100
·期 7
·3062-3070
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IF 2.614
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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APPLIED AND ENVIRONMENTAL MICROBIOLOGY
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2015
·Vol. 81
·期 6
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IF 3.668
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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Nature Communications
·
2025
·Vol. 16
·期 1
·1-14
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IF 15.7
Fan Rong,Guan Zeyuan,Zhou Guanghong,Yang Xi,Zhang Fei,Wu Menglong,Wang Xuecui,Liu Jian,Chen Pei,Liu Yanjun,Zhang Delin,Yin Ping,Yan Junjie
Amylopectin, the primary form of starch in plant leaves, seeds and tubers, features a tree-like architecture with branched glucose chains. Excess branches result in the formation of soluble phytoglycogen instead of starch granules. In higher plants and green algae, the debranching enzyme isoamylase ISA1 forms either homomultimer or hetero-multimer with ISA2 to facilitate branch trimming and starch granule formation, but the molecular basis remains largely unknown. In this study, we reconstitute the rice OsISA1-ISA2 complex in vitro and determine the cryo-EM structures of the OsISA1 homodimer, as well as the malto-oligosaccharide (MOS)-free and MOS-bound OsISA1-ISA2 heterocomplex. The OsISA1 dimer shows a tail-to-tail rod-like architecture, whereas the OsISA1-ISA2 complex mainly exhibits as a trimer, with OsISA2 flanking on the N-terminal segments of the dimeric OsISA1. Combined with comprehensive biochemical analyses, these structural data elucidate the organization of the ISA1-ISA2 heterocomplex in higher plants and demonstrate how ISA1 and ISA2 cooperate during amylopectin biosynthesis.
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Journal of Advanced Research
·
2025
·
IF 13
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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危险品化学品经营许可证（带存储）