Molecular Catalysis
·
2022
·
IF 5.089
Hengtao Tang,Zheng Zhou,Zhi Chen,Xin Ju,Liangzhi Li
Monosaccharides from biomass can be used as cost-effective raw materials to produce rare sugars which are valuable sweeteners. Enzymes play a pivotal role in such biomass biorefinery processes to develop novel synthetic routes, and improve the environmental friendliness of current routes. Here we propose a proof of concept biorefinery process to prepare useful rare sugars of D-xylulose and D-ribulose from D-xylose which was abundant in biomass. Ribose-5-phosphate isomerase B from Ochrobactrum sp. (OsRpiB) with a unique substrate scope was engineered to intensify the activity of transforming D-xylose to D-xylulose. A double mutant R94N/N137Q with 80% higher catalytic efficiency was identified, and kinetic and molecular dynamic studies were conducted to explore the mechanism underneath. The results revealed a correlation between enhancing stability and activity simultaneously. Then a one-pot enzyme cascade was constructed with OsRpiB and D-tagatose-3-epimerase from Pseudomonas cichorii and transformed D-xylulose further to D-ribulose. The conversions of D-xylose and D-xylulose in cascade reaction were 1.80-fold and 1.83-fold of that in separate reactions with the aid of process parameter adjustment. The presented work is the first reported enzyme cascade converting D-xylose to D-ribulose, and proves the promising role of monosaccharide isomerase platforms in the refinery of biomass.
研究领域:
生物化学
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酶工程
·
生物质精炼
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产品关联
DOI
高影响因子
BIOTECHNOLOGY LETTERS
·
2023
·Vol. 45
·期 10
·1355-1364
·
IF 2.7
Gao Yahui,Li Fen,Wang Yulu,Chen Zhou,Li Zijie
Purpose In our previous study, we constructed a one-pot multi-enzyme system for rare ketoses synthesis based on L-rhamnulose-1-phosphate aldolase (RhaD) from accessible glycerol in vitro. To eliminate tedious purification of enzymes, a facile Escherichia coli whole-cell cascade platform was established in this study. Methods To enhance the conversion rate, the reaction conditions, substrate concentrations and expressions of related enzymes were extensively optimized. Results The biosynthetic route for the cascade synthesis of rare ketoses in whole cells was successfully constructed and three rare ketoses including D-allulose, D-sorbose and L-fructose were produced using glycerol and D/L-glyceraldehyde (GA). Under optimized conditions, the conversion rates of rare ketoses were 85.0% and 93.0% using D-GA and L-GA as the receptor, respectively. Furthermore, alditol oxidase (AldO) was introduced to the whole-cell system to generate D-GA from glycerol, and the total production yield of D-sorbose and D-allulose was 8.2 g l −1 only from the sole carbon source glycerol. Conclusion This study demonstrates a feasible and cost-efficient method for rare sugars synthesis and can also be applied to the green synthesis of other value-added chemicals from glycerol.
研究领域:
生物化学
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合成生物学
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代谢工程
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DOI
高影响因子
CHEMICAL ENGINEERING SCIENCE
·
2021
·
IF 4.311
Xiaoyan Chen,Fan Wang,Qiang Yu,Shijie Liu,Wen Wang,Yu Zhang,Zhongming Wang,Zhenhong Yuan
One pot cascade of enzymatic catalysis was applied for d -allulose synthesis from d -glucose by the combination of d -glucose isomerase (GI) from Escherichia coli MG1655 and d -allulose 3-epimerase (DAE) from Agrobacterium tumefaciens C58. The synthesis process was investigated by characterizing the initial substrate concentration, molar ratios of GI and DAE, the enzyme loading, and contents of cobalt and magnesium ions. The increase of enzyme loading and the molar ratios of GI vs DAE caused the raising of d -allulose yield from 4% to 9%, verifying that the generation rate of d -fructose was an essential factor for rare sugar conversion improvement. The optimal enzyme loadings required for reactions with substrate concentrations of 100 g L −1 and 200 g L −1 were 60 U and 200 U. Kinetics stimulation was conducted to reveal the underlying relationship within the three sugars converted by dual-enzyme, with the fitting curve validated the reliability of the model.
研究领域:
生物化学
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酶工程
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食品科学
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产品关联
DOI
高影响因子
BIORESOURCE TECHNOLOGY
·
2016
·
IF 4.917
Hongsen Zhang,Gang Liu,Jian Zhang,Jie Bao
High titer gluconic acid and xylonic acid were simultaneously fermented by Gluconobacter oxydans DSM 2003 using corn stover feedstock after dry dilute sulfuric acid pretreatment, biodetoxification and high solids content hydrolysis. Maximum sodium gluconate and xylonate were produced at the titer of 132.46 g/L and 38.86 g/L with the overall yield of 97.12% from glucose and 90.02% from xylose, respectively. The drawbacks of filamentous fungus Aspergillus niger including weak inhibitor tolerance, large pellet formation and no xylose utilization were solved by using the bacterium strain G. oxydans . The obtained sodium gluconate/xylonate product was highly competitive as cement retarder additive to the commercial product from corn feedstock. The techno-economic analysis (TEA) based on the Aspen Plus modeling was performed and the minimum sodium gluconate/xylonate product selling price (MGSP) was calculated as $0.404/kg. This study provided a practical and economic competitive process of lignocellulose utilization for production of value-added biobased chemicals.
研究领域:
生物工程
·
可再生资源利用
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产品关联
DOI
PubMed
高影响因子
ADVANCED SYNTHESIS & CATALYSIS
·
2016
·Vol. 358
·期 24
·4030-4040
·
IF 6.453
Lingtian Wu,Juanjuan Qiu,Shanshan Wu,Xiaoliu Liu,Chao Liu,Zheng Xu,Sha Li,Hong Xu
Isomaltulose is a new functional sweetener that possesses biological characteristics and can be obtained by enzymatic synthesis using sucrose as substrate. In this study, the sucrose isomerase (SIase) was successfully immobilized on the sponge synthesized with ϵ-poly- l -lysine and gelatin. The inhibition zones proved that the sponge possessed a significant bacteriostatic function, which can effectively prevent the collapse of the scaffolds of the immobilized enzymes. The immobilized SIase activity reached up to 71.58 U g −1 (the SIase loading was about 153.43 mg g −1 ), and the activity recovery was approximately 84.50%. After immobilization, the optimum pH of SIase decreased from 6.0 to 5.5 and the optimum temperature increased from 30 °C to 40 °C. The affinity of SIase to substrate was basically unchanged. Immobilized SIase still exhibited more than 90% sucrose conversion after 13 consecutive cycles, which indicated that it had a good operational stability. Furthermore, the immobilized SIase has the potential for isomaltulose production, with 200 g L −1 sucrose solution as its substrate in the food industry. Isomaltulose was isolated in 83.58% yield and high purity (97.3%).
研究领域:
生物化学
·
食品科学
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产品关联
DOI
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2025
·
IF 3.7
Chen-Xin Guo,Jiyan Li,Yu Wang,Tang Li,Heng Yin
Stevia rebaudiana , a perennial herb, is recognized not only for its sweet steviol glycosides but also for its rich flavonoid content, which confer pharmacological properties including anti-inflammatory, antimicrobial, and anticancer activities. However, the enzymatic basis underlying flavonoid modification in S. rebaudiana remains poorly understood. In this study, we identified, cloned, and heterologously expressed a novel flavonoid glycosyltransferase gene, SrUGT72B1 in E. coli . The recombinant SrUGT72B1 catalyzed the glycosylation of multiple flavonoids using UDP-glucose as the primary sugar donor, and exhibited broad substrate promiscuity toward apigenin, luteolin, phloretin and kaempferol. In addition to UDP-glucose, SrUGT72B1 also accepted UDP-xylose and UDP-rhamnose, with UDP-glucose exhibiting the highest catalytic efficiency. Biochemical characterization revealed that the enzyme functions optimally at pH 9.0 and 50 °C. Notably, SrUGT72B1 demonstrates regioselective 5-O-glycosylation toward apigenin, a rare activity among plant glycosyltransferases. Molecular docking and molecular dynamics simulations provided structural insights into this unique regioselectivity and substrate recognition. Together, these findings establish SrUGT72B1 as a previously uncharacterized flavonoid 5-O-glycosyltransferase, expanding the functional landscape of plant UGTs and offering potential applications in the biosynthesis of value-added flavonoid glycosides.
研究领域:
植物学
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生物化学
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分子生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
·
2024
·
IF 3.3
Zirou Xiao,Shuang Wu,Hongshan Liang,Bin Li,Jing Li
BACKGROUND Growing public concern over the health risks of high-sugar diets has led to a consensus on the necessity of sugar reduction. This research evaluated the effects of oligosaccharides (OS), sodium carboxymethyl cellulose (CMCNa) and d -allulose as a compound improver on the quality of sugar-reduced bread, aiming to assess the feasibility of substantial sucrose reduction at the same time as maintaining desirable bread characteristics. RESULTS Compared to bread with a 90% sugar reduction, the improved formulation increased specific volume by 50.14% and reduced hardness by 66.69%. It exhibited a homogeneous structure and crust color closely resembling that of full-sugar bread. Low-field NMR analysis revealed better moisture control, delaying water loss and starch retrogradation, with relative crystallinity and retrogradation enthalpy decreased by 45.31% and 59.96%, respectively. Additionally, the combination of XOS and d -allulose boosted volatile compound production, increasing the abundance of aldehydes, esters and heterocyclic compounds, enriching the flavor with fruity and baked aromas. CONCLUSION The improvers enhanced the texture, appearance, flavor and storage stability of sugar-reduced bread, yielding qualities that are comparable to or even surpass conventional bread. These findings provide a new insight for the development of quality improvers designed for sugar-reduced bakery products. © 2024 Society of Chemical Industry.
研究领域:
食品科学
·
营养学
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产品关联
DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2025
·
IF 8
Zirou Xiao,Shuang Wu,Hongshan Liang,Bin Li,Jing Li
The reduction of sucrose in dough formulations disrupts mechanical properties, fermentation performance, and structural organization, leading to rigidity and impaired gas retention. This study investigated how composite improvers—oligosaccharides (XOS/KOS), CMCNa, and D-allulose—mitigate these challenges by restoring viscoelastic balance, enhancing fermentation, and stabilizing protein and starch interactions. Correlation analysis was further conducted to explore the complex relationships between these parameters. Results revealed that D-allulose optimized gas retention by contributing to a denser gluten network, achieving the highest retention rate of 90.92 %. Oligosaccharides improved water availability, promoting gluten development and enhancing starch gelatinization, while CMCNa strengthened the gluten network through its binding and film-forming properties, collectively increasing fermentation height by 79.78 % compared to sugar-reduced dough. These structural improvements were further confirmed by SEM images, which revealed a more uniform microstructure with continuous gluten films encapsulating starch granules. These findings elucidate the mechanisms underlying the deterioration of sugar-reduced dough and highlight the potential of composite improvers in mitigating the adverse effects of sucrose reduction, paving the way for the development of high-quality sugar-reduced products.
研究领域:
食品科学
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生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
·
2022
·Vol. 38
·期 8
·1-11
·
IF 4.253
Xue Kai,Liu Chun-Li,Yang Yankun,Liu Xiuxia,Zhan Jinling,Bai Zhonghu
d -allulose is a rare low-calorie sugar that has many fundamental biological functions. d -allulose 3-epimerase from Agrobacterium tumefaciens (AT-DAEase) catalyzes the conversion of d -fructose to d -allulose. The enzyme has attracted considerable attention because of its mild catalytic properties. However, the bioconversion efficiency and reusability of AT-DAEase limit its industrial application. Magnetic metal–organic frameworks (MOFs) have uniform pore sizes and large surface areas and can facilitate mass transport and enhance the capacity for enzyme immobilization. Here, we successfully encapsulated cobalt-type AT-DAEase into the cobalt-based magnetic MOF ZIF-67@Fe 3 O 4 using a self-assembly strategy. We confirmed the immobilization of enzyme AT-DAEase and characterized the enzymatic properties of the MOF-immobilized AT-DAEase@ZIF-67@Fe 3 O 4 . The AT-DAEase@ZIF-67@Fe 3 O 4 nanoparticles had higher catalytic activity (65.1 U mg −1 ) and bioconversion ratio (38.1%) than the free AT-DAEase. The optimal conditions for maximum enzyme activity of the AT-DAEase@ZIF-67@Fe 3 O 4 nanoparticles were 55 °C and pH 8.0, which were significantly higher than those of the free AT-DAEase (50 °C and pH 7.5). The AT-DAEase@ZIF-67@Fe 3 O 4 nanoparticles displayed significantly improved thermal stability and excellent recycling performance, with 80% retention of enzyme activity at a temperature range of 45–70 °C and > 45% of its initial activity after eight cycles of enzyme use. The AT-DAEase@ZIF-67@Fe 3 O 4 nanoparticles have great potential for large-scale industrial preparation of d -allulose by immobilizing cobalt-type AT-DAEase into magnetic MOF ZIF-67@Fe 3 O 4 .
研究领域:
生物化学
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食品科学
·
材料科学
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产品关联
DOI
PubMed
高影响因子
Plant Methods
·
2019
·Vol. 15
·期 1
·1-13
·
IF 3.17
Song Yadi,Chen Shang,Wang Xiujuan,Zhang Rui,Tu Lichan,Hu Tianyuan,Liu Xihong,Zhang Yifeng,Huang Luqi,Gao Wei
Background Tripterygium wilfordii Hook. f. ( T. wilfordii ) is an important medicinal plant with anti-inflammatory, immunosuppressive and anti-tumor activities. The main bioactive ingredients are diterpenoids and triterpenoids, such as triptolide, triptophenolide and celastrol. However, the production of terpenoids from original plants, hairy roots and dedifferentiated cells (DDCs) are not satisfactory for clinical applications. To find a new way to further improve the production of terpenoids, we established a new culture system of cambial meristematic cells (CMCs) with stem cell-like properties, which had strong vigor and high efficiency to produce large amounts of terpenoids of T. wilfordii . Results CMCs of T. wilfordii were isolated and cultured for the first time. CMCs were characterized consistent with stem cell identities based on their physiological and molecular analysis, including morphology of CMCs, hypersensitivity to zeocin, thin cell wall and orthogonal partial least square-discriminant analysis, combination of transcriptional data analysis. After induction with methyl jasmonate (MJ), the maximal production of triptolide, celastrol and triptophenolide in CMCs was 312%, 400% and 327% higher than that of control group, respectively. As for medium, MJ-induced CMCs secreted 231% triptolide and 130% triptophenolide at the maximum level into medium higher than that of control group. Maximal celastrol production of induced CMCs medium was 48% lower than that of control group. Long-term induction significantly enhanced the production of terpenoids both in cells and medium. The reason for increasing the yield of terpenoids was that expression levels of 1 - deoxy - d - xylulose - 5 - phosphate synthase ( DXS ), 1 - deoxy - d - xylulose - 5 - phosphate reductoisomerase ( DXR ) and hydroxymethylglutaryl - CoA synthase ( HMGS ) were upregulated in CMCs after induction. Conclusions For the first time, CMCs of T. wilfordii were isolated, cultured, characterized and applied. Considering the significant enrichment of terpenoids in CMCs of T. wilfordii , CMCs could provide an efficient and controllable platform for sustainable production of terpenoids, which can be a better choice than DDCs.
研究领域:
植物生物技术
·
植物化学
·
药用植物
✓ 已验证
DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
·
2021
·
IF 5.89
Wang Zhaoyu,Li Yang,Li Mingyi,Zhang Xiaohui,Ji Qingxia,Zhao Xiaojuan,Bi Yanhong,Luo Si
The conversion of flavonoid glycosides and their analogs to their lipophilic ester derivatives was developed by nanobiocatalysts from immobilizing Thermomyces lanuginosus lipase (TLL) on polydopamine-functionalized magnetic Fe3O4 nanoparticles (Fe3O4-PDA-TLL). The behavior investigation revealed that Fe3O4-PDA-TLL exhibits a preference for long chain length fatty acids (i.e., C10 to C14) with higher reaction rates of 12.6–13.9 mM/h. Regarding the substrate specificity, Fe3O4-PDA-TLL showed good substrate spectrum and favorably functionalized the primary OH groups, suggesting that the steric hindrances impeded the secondary or phenolic hydroxyl groups of substrates into the bonding site of the active region of TLL to afford the product.
研究领域:
生物化学
·
酶工程
·
有机化学
✓ 已验证
DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2024
·
IF 3.4
Dingyu Guo,Zhengchao Wang,Wanqing Wei,Wei Song,Jing Wu,Jian Wen,Guipeng Hu,Xiaomin Li,Cong Gao,Xiulai Chen,Liming Liu
D-allulose is a naturally occurring rare sugar and beneficial to human health. However, the efficient biosynthesis of D-allulose remains a challenge. Here, we mined a new D-tagatose 3-epimerase from Kroppenstedtia eburnean (KeDt3e) with high catalytic efficiency . Initially, crucial factors contributing to the low conversion of KeDt3e were identified through crystal structure analysis, density functional theory calculations (DFT), and molecular dynamics (MD) simulations. Subsequently, based on the mechanism, combining restructuring the flexible region, proline substitution based onconsensus sequence analysis, introducing disulfide bonds , and grafting properties, and reshaping the active center, the optimal mutant M5 of KeDt3e was obtained with enhanced thermostability and activity. The optimal mutant M5 exhibited an enzyme activity of 130.8 U/mg, representing a 1.2-fold increase; T m value increased from 52.7 °C to 71.2 °C; and half-life at 55 °C extended to 273.7 min, representing a 58.2-fold improvement, and the detailed mechanism of performance improvement was analyzed. Finally, by screening the ribosome-binding site (RBS) of the optimal mutant M5 recombinant bacterium (G01), the engineered strain G08 with higher expression levels was obtained. The engineered strain G08 catalyzed 500 g/L D-fructose to produce 172.4 g/L D-allulose, with a conversion of 34.4% in 0.5 h and productivity of 344.8 g/L/h on a 1 L scale. This study presents a promising approach for industrial-scale production of D-allulose.
研究领域:
生物化学
·
食品科学
·
酶工程
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）