JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2026
·Vol. 74
·期 30
·23649-23661
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IF 6.7
Huang Haidong,Li Yangyang,Song Jinsong,Zhao Yuhan,Liu Yanfeng,Li Jianghua,Du Guocheng,Chen Jian,Li Zhendong,Lv Xueqin,Xu Xianhao,Liu Long
Trehalose is a nonreducing disaccharide widely used for
its biomolecule-protective
properties. However, multienzyme cascade production remains limited
by low enzyme expression and suboptimal catalytic performance. To
address this, thermostable maltooligosyltrehalose synthase (TreY)
and trehalohydrolase (TreZ) from Arthrobacter ramosus were individually expressed intracellularly in Bacillus subtilis , and the crude lysates were combined for
trehalose biosynthesis, achieving 281.4 g/L trehalose and a yield
of 0.7 g trehalose/g maltodextrin. Integrated computational screening
identified MalQ-3 from Cyanobacterium stanieri as a suitable 4-α-glucanotransferase for soluble expression.
Subsequent semirational engineering generated MalQ-3-M2 (S54P/V472F),
with enhanced activity and stability associated with improved substrate-pocket
dynamics, thereby facilitating glucan-chain rearrangement and short-chain
reutilization. MalQ-3-M2 was
separately expressed in B. subtilis and incorporated into the crude-lysate cascade, increasing the trehalose
titer to 338 g/L and the yield to 0.85 g trehalose/g maltodextrin.
Overall, this work establishes a scalable B. subtilis platform for efficient trehalose production.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2026
·
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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JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2023
·
IF 3.4
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
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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FOOD CHEMISTRY
·
2020
·
IF 6.306
Yu Lu,Cheng Li,Jie Liu,Wenqi Huang,Yuerong Yang,Yue Jia,Tingting Liu,Linjuan Huang,Zhongfu Wang
Goat milk oligosaccharides represent an unexplored multi-functional ingredient for the dairy industry. Here, we qualitatively and quantitatively compared the N/O -glycome at different lactation stages via online hydrophilic interaction chromatography-tandem mass spectrometry. Complex N -glycans and high mannose N -glycans constituted 82.1% and 17.9% of the glycan pool, respectively. N -glycans with isomers containing non-bisected antenna complex structures accounted for 30.8%. N -glycans modified with Neu5Ac, Neu5Gc and fucosylated were 3.7%, 5.3% and 35.3%. The triantennary trifucosylated complex N -glycan (H5N5F3) was reported for the first time. A comparison between colostrum and mature milk revealed a 1.20-fold decrease in total N -glycans and 1.66-fold decrease in fucosylation with ongoing lactation, echoing the trend in human milk. Similarly, Neu5Ac- and Neu5Gc-modified sialylation decreased by 1.69 and 3.62 times, respectively. In the O -glycome, 46.2% of structures were O -linked core 1, 23.1% were O -linked core 2, 7.7% were O -linked core 3 and core 4. As lactation progressed, overall O -glycans content decreased by 1.26-fold. Unlike human milk, Neu5Ac- and Neu5Gc-modified sialylation increased by 4.4 and 2 times, respectively. These findings will facilitate research on the structure-function relationship of goat milk oligosaccharides and the development of formula food targeting different age groups.
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JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
·
2020
·Vol. 100
·期 7
·3062-3070
·
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
·
2015
·Vol. 81
·期 6
·
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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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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