Enhancing Glycosylation of Flavonoids by Engineering the Uridine Diphosphate Glucose Supply in <i>Escherichia coli</i>

Liu, Shike; Li, Dong; Qin, Zhijie; Zeng, Weizhu; Zhou, Jingwen

doi:10.1021/acs.jafc.3c05264

Cited by 8 publications

(3 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The expression levels of exogenous genes need to be optimized to achieve the highest 5-MTHF titer. An appropriate gene copy number is a significant factor, as excessively high copy number might burden host growth, while low copy number might reduce expression levels, both of which are not conducive to the accumulation of 5-MTHF . Therefore, we selected five different replicons with varying copy numbers: pSC101 (low), ColA (low), p15A (medium), pBR322 (medium), and pMB1 (derivative) (high) (Table S3).…”

Section: Resultsmentioning

confidence: 99%

Engineering Redox Cofactor Balance for Improved 5-Methyltetrahydrofolate Production in Escherichia coli

Yang,

Wu,

et al. 2024

J. Agric. Food Chem.

View full text Add to dashboard Cite

5-Methyltetrahydrofolate (5-MTHF) is the sole active form of folate functioning in the human body and is widely used as a nutraceutical. Unlike the pollution from chemical synthesis, microbial synthesis enables green production of 5-MTHF. In this study, Escherichia coli BL21 (DE3) was selected as the host. Initially, by deleting 6-phosphofructokinase 1 and overexpressing glucose-6-phosphate 1-dehydrogenase and 6-phosphogluconate dehydrogenase, the glycolysis pathway flux decreased, while the pentose phosphate pathway flux enhanced. The ratios of NADH/NAD+ and NADPH/NADP+ increased, indicating elevated NAD(P)H supply. This led to more folate being reduced and the successful accumulation of 5-MTHF to 44.57 μg/L. Subsequently, formate dehydrogenases from Candida boidinii and Candida dubliniensis were expressed, which were capable of catalyzing the reaction of sodium formate oxidation for NAD(P)H regeneration. This further increased the NAD(P)H supply, leading to a rise in 5-MTHF production to 247.36 μg/L. Moreover, to maintain the balance between NADH and NADPH, pntAB and sthA, encoding transhydrogenase, were overexpressed. Finally, by overexpressing six key enzymes in the folate to 5-MTHF pathway and employing fed-batch cultivation in a 3 L fermenter, strain Z13 attained a peak 5-MTHF titer of 3009.03 μg/L, the highest level reported in E. coli so far. This research is a significant step toward industrial-scale microbial 5-MTHF production.

show abstract

Section: Resultsmentioning

confidence: 99%

Engineering Redox Cofactor Balance for Improved 5-Methyltetrahydrofolate Production in Escherichia coli

Yang,

Wu,

et al. 2024

J. Agric. Food Chem.

View full text Add to dashboard Cite

show abstract

“…Optimization of UDPG supply is a commonly used tool in glycosylation pathway construction [ 47 ]. In S. cerevisiae , glucose is first converted to glucose 6-phosphate (G-6-P), then to glucose 1-phosphate (G-1-P) by the action of PGM1/PGM2 , and UDPG is finally catalyzed by UGP1 [ 22 ].…”

Section: Discussionmentioning

confidence: 99%

Systematic Engineering of Saccharomyces cerevisiae for the De Novo Biosynthesis of Genistein and Glycosylation Derivatives

Wang,

Xiao,

Zhang

et al. 2024

JoF

View full text Add to dashboard Cite

Isoflavones are predominantly found in legumes and play roles in plant defense and prevention of estrogen-related diseases. Genistein is an important isoflavone backbone with various biological activities. In this paper, we describe how a cell factory that can de novo synthesize genistein was constructed in Saccharomyces cerevisiae. Different combinations of isoflavone synthase, cytochrome P450 reductase, and 2-hydroxyisoflavone dehydratase were tested, followed by pathway multicopy integration, to stably de novo synthesize genistein. The catalytic activity of isoflavone synthase was enhanced by heme supply and an increased intracellular NADPH/NADP+ ratio. Redistribution of the malonyl-CoA flow and balance of metabolic fluxes were achieved by adjusting the fatty acid synthesis pathway, yielding 23.33 mg/L genistein. Finally, isoflavone glycosyltransferases were introduced into S. cerevisiae, and the optimized strain produced 15.80 mg/L of genistin or 10.03 mg/L of genistein-8-C-glucoside. This is the first de novo synthesis of genistein-8-C-glucoside in S. cerevisiae, which is advantageous for the green industrial production of isoflavone compounds.

show abstract

“…Since the backbone of flavonoid is based on a 15-carbon skeleton with two phenyl rings connected by a heterocyclic ring (ring C), the biosynthetic pathway of diversified flavonoids requires various types of functional enzymes. The modified derivatives of flavonoids are synthesized by methylation, hydroxylation, glycosylation, isoprenylation, and halogenation [63,64]. The diversified flavonoids showed improved bioavailability compared with flavonoid aglycone.…”

Section: Improving Intracellular Udp-sugar Level For Biosynthesis Of ...mentioning

confidence: 99%

Metabolic Engineering of Corynebacterium glutamicum for the Production of Flavonoids and Stilbenoids

Chu,

Tran,

Pham

et al. 2024

Molecules

View full text Add to dashboard Cite

Flavonoids and stilbenoids, crucial secondary metabolites abundant in plants and fungi, display diverse biological and pharmaceutical activities, including potent antioxidant, anti-inflammatory, and antimicrobial effects. However, conventional production methods, such as chemical synthesis and plant extraction, face challenges in sustainability and yield. Hence, there is a notable shift towards biological production using microorganisms like Escherichia coli and yeast. Yet, the drawbacks of using E. coli and yeast as hosts for these compounds persist. For instance, yeast’s complex glycosylation profile can lead to intricate protein production scenarios, including hyperglycosylation issues. Consequently, Corynebacterium glutamicum emerges as a promising alternative, given its adaptability and recent advances in metabolic engineering. Although extensively used in biotechnological applications, the potential production of flavonoid and stilbenoid in engineered C. glutamicum remains largely untapped compared to E. coli. This review explores the potential of metabolic engineering in C. glutamicum for biosynthesis, highlighting its versatility as a cell factory and assessing optimization strategies for these pathways. Additionally, various metabolic engineering methods, including genomic editing and biosensors, and cofactor regeneration are evaluated, with a focus on C. glutamicum. Through comprehensive discussion, the review offers insights into future perspectives in production, aiding researchers and industry professionals in the field.

show abstract

Enhancing Glycosylation of Flavonoids by Engineering the Uridine Diphosphate Glucose Supply in Escherichia coli

Cited by 8 publications

References 39 publications

Engineering Redox Cofactor Balance for Improved 5-Methyltetrahydrofolate Production in Escherichia coli

Engineering Redox Cofactor Balance for Improved 5-Methyltetrahydrofolate Production in Escherichia coli

Systematic Engineering of Saccharomyces cerevisiae for the De Novo Biosynthesis of Genistein and Glycosylation Derivatives

Metabolic Engineering of Corynebacterium glutamicum for the Production of Flavonoids and Stilbenoids

Contact Info

Product

Resources

About