Artificially Created UDP-Glucose 2-Epimerase Enables Concise UDP/GDP-Mannose Production via the Synthase–Epimerase Route DOI

Zhongbao Ma,

Liting Zhao,

Qiong Wang

и другие.

ACS Catalysis, Год журнала: 2024, Номер unknown, С. 18072 - 18084

Опубликована: Ноя. 22, 2024

Uridine/guanosine diphosphate-mannose (UDP/GDP-Man) is the major mannosyl donor in producing mannose-containing oligo/polysaccharides. Its acquisition greatly limited by its complex and costly synthetic process, which requires multiple substrates enzymes. The natural UDP/GDP-glucose 2-epimerase functioning C2 epimerization between UDP/GDP-Glc UDP/GDP-Man remains unreported main hurdle to realize concise production of UDP/GDP-Man. Here, UDP-glucose (Glc2E), behaves like a naturally evolved enzyme, created exhibits high-efficient catalysis UDP-Man. Multidimensional engineering, including redesigning nucleobase recognition region, displacement substrate tunnel entrance, expansion space for sugar ring rotation, employed develop Glc2E from CDP-tyvelose 2-epimerase. converts 55.63% UDP-Glc UDP-Man, trace value initial stTyvE, aptitude GDP-Glc evolves unobserved activity 23.94% conversion. Coupling sucrose synthase with achieves theoretical synthase–epimerase route inexpensive sucrose. space-time-yield UDP-Man maximized 8.05 g/L/h within 2.5 h, final titer 22.54 g/L, demonstrating competitive application potential. Moreover, GDP-Man synthesized successfully at 3.49 g/L. Our work inspires enzyme engineering epimerases glycosyltransferases that catalyze nucleotide sugars. unlocks feasible approach cost-competitive donors.

Язык: Английский

Metabolic engineering of Escherichia coli BL21(DE3) cocultured with glucose and xylose for efficient production of 2′-fucosyllactose DOI
Yunqi Zhu,

Mingli Zhao,

Hao Wang

и другие.

Bioresource Technology, Год журнала: 2025, Номер unknown, С. 132062 - 132062

Опубликована: Янв. 1, 2025

Язык: Английский

Процитировано

3
Highly-efficient in vivo production of lacto-N-fucopentaose V by a regio-specific α1,3/4-fucosyltransferase from Bacteroides fragilis NCTC 9343 DOI
Ningning Wang, Yingying Zhu, Liang Wang

и другие.

International Journal of Biological Macromolecules, Год журнала: 2024, Номер 266, С. 130955 - 130955

Опубликована: Март 16, 2024

Язык: Английский

Процитировано

5
Construction and application of 3-fucosyllactose whole-cell biosensor for high-throughput screening of overproducers DOI
Qinggang Li, Chuan Liu,

Jinhuai He

и другие.

Bioresource Technology, Год журнала: 2024, Номер 402, С. 130798 - 130798

Опубликована: Май 3, 2024

Язык: Английский

Процитировано

4
Recent advances of 3-fucosyllactose in health effects and production DOI
Zhihui Du, Zeyu Li,

Cuie Guang

и другие.

Archives of Microbiology, Год журнала: 2024, Номер 206(9)

Опубликована: Авг. 14, 2024

Язык: Английский

Процитировано

4
Highly Efficient In Vivo Production of Sialyllacto-N-tetraose C via Screening of Beneficial β1,4-galactosyltransferase and α2,6-sialyltransferase DOI
Yuanlin Liu,

Qian Lin,

Mian Sheng

и другие.

Journal of Agricultural and Food Chemistry, Год журнала: 2025, Номер 73(9), С. 5376 - 5384

Опубликована: Фев. 18, 2025

Biological production of human milk oligosaccharides (HMOs) using metabolically engineered strains is a research hotspot in food biotechnology, but less effort has been made on the biological sialylated complex HMOs. Sialyllacto-N-tetraose c only monosialylated HMO top 15 In this study, metabolic pathway LST was constructed Escherichia coli BL21(DE3) by introducing three sequential glycosyltransferases: β1,3-N-acetylglucosaminyltransferase, β1,4-galactosyltransferase, and α2,6-sialyltransferase. The cytidine 5′-monophospho (CMP)-N-acetylneuraminic acid (Neu5Ac) enhanced to improve production. β1,4-galactosyltransferase from Helicobacter pylori J99 (HpGalT) α2,6-sialyltransferase Vespertiliibacter pulmonis (ED6ST) were screened as pair key glycosyltransferases for enhancing final strain could produce 1.718 9.745 g/L shake-flask fed-batch cultivation, respectively, indicating feasibility efficient biosynthesis

Язык: Английский

Процитировано

0
Structure, Function, Synthesis and Improved Strategies of Fucosylated Human Milk Oligosaccharides and Their Future Perspectives: A Review DOI

Ya-Ya Yang,

Shunli Jing,

Le Zhang

и другие.

Food Bioscience, Год журнала: 2025, Номер unknown, С. 106584 - 106584

Опубликована: Апрель 1, 2025

Язык: Английский

Процитировано

0
Glycosyltransferases: glycoengineers in human milk oligosaccharide synthesis and manufacturing DOI Creative Commons

A. Slater,

Andrew G. McDonald, Rita M. Hickey

и другие.

Frontiers in Molecular Biosciences, Год журнала: 2025, Номер 12

Опубликована: Апрель 30, 2025

Human milk oligosaccharides (HMOs) are a diverse group of complex carbohydrates that play crucial roles in infant health, promoting beneficial gut microbiota, modulating immune responses, and protecting against pathogens. Central to the synthesis HMOs glycosyltransferases, specialized class enzymes catalyse transfer sugar moieties form glycan structures characteristic HMOs. This review provides an in-depth analysis beginning with their classification based on structural functional characteristics. The catalytic activity these is explored, highlighting mechanisms by which they facilitate precise addition monosaccharides HMO biosynthesis. Structural insights into glycosyltransferases also discussed, shedding light how conformational features enable specific glycosidic bond formations. maps out key biosynthetic pathways involved production, including lactose, subsequent fucosylation sialylation processes, all intricately regulated glycosyltransferases. Industrial methods for synthesis, chemical, enzymatic, microbial approaches, examined, emphasizing role processes. Finally, discusses future directions glycosyltransferase research, particularly enhancing efficiency developing advanced analytical techniques better understand complexity biological functions

Язык: Английский

Процитировано

0
Glycosyltransferases in human milk oligosaccharide synthesis: structural mechanisms and rational design DOI
Yanfeng Liu, Guocheng Du, Jian Chen

и другие.

Current Opinion in Biotechnology, Год журнала: 2025, Номер 93, С. 103315 - 103315

Опубликована: Май 9, 2025

Язык: Английский

Процитировано

0
Characterization of a GH20 β-N-Acetylhexosaminidase from Flavobacterium algicola Suitable to Synthesize Lacto-N-triose II DOI
Chengqiang Li,

Zhuoning Cao,

Hong Jiang

и другие.

Journal of Agricultural and Food Chemistry, Год журнала: 2024, Номер 72(9), С. 4849 - 4857

Опубликована: Фев. 22, 2024

β-N-Acetylhexosaminidases have attracted much attention in the enzymatic synthesis of lacto-N-triose II (LNT2) as a backbone precursor human milk oligosaccharides (HMOs). In this study, novel glycoside hydrolase (GH) 20 family β-N-acetylhexosaminidase, FlaNag2353, from Flavobacterium algicola was biochemically characterized and applied to synthesize LNT2. FlaNag2353 displayed optimal activity p-nitrophenyl N-acetyl-β-d-glucosaminide (pNP-GlcNAc) at 40 °C pH 8.0. addition its excellent hydrolysis toward pNP-GlcNAc chitooligosaccharides, showed trans-glycosylation activity. Under conditions 9.0 55 for 2 h utilizing 200 mM lactose 10 pNP-GlcNAc, synthesized LNT2 with conversion ratio 4.15% calculated pNP-GlcNAc. Moreover, when 9.7% (w/v) industrial waste whey powder, achieved 2.39%. This study has significant implications broadening applications GH20 β-N-acetylhexosaminidases promoting high-value utilization powder.

Язык: Английский

Процитировано

2
Artificially Created UDP-Glucose 2-Epimerase Enables Concise UDP/GDP-Mannose Production via the Synthase–Epimerase Route DOI

Zhongbao Ma,

Liting Zhao,

Qiong Wang

и другие.

ACS Catalysis, Год журнала: 2024, Номер unknown, С. 18072 - 18084

Опубликована: Ноя. 22, 2024

Uridine/guanosine diphosphate-mannose (UDP/GDP-Man) is the major mannosyl donor in producing mannose-containing oligo/polysaccharides. Its acquisition greatly limited by its complex and costly synthetic process, which requires multiple substrates enzymes. The natural UDP/GDP-glucose 2-epimerase functioning C2 epimerization between UDP/GDP-Glc UDP/GDP-Man remains unreported main hurdle to realize concise production of UDP/GDP-Man. Here, UDP-glucose (Glc2E), behaves like a naturally evolved enzyme, created exhibits high-efficient catalysis UDP-Man. Multidimensional engineering, including redesigning nucleobase recognition region, displacement substrate tunnel entrance, expansion space for sugar ring rotation, employed develop Glc2E from CDP-tyvelose 2-epimerase. converts 55.63% UDP-Glc UDP-Man, trace value initial stTyvE, aptitude GDP-Glc evolves unobserved activity 23.94% conversion. Coupling sucrose synthase with achieves theoretical synthase–epimerase route inexpensive sucrose. space-time-yield UDP-Man maximized 8.05 g/L/h within 2.5 h, final titer 22.54 g/L, demonstrating competitive application potential. Moreover, GDP-Man synthesized successfully at 3.49 g/L. Our work inspires enzyme engineering epimerases glycosyltransferases that catalyze nucleotide sugars. unlocks feasible approach cost-competitive donors.

Язык: Английский

Процитировано

1