Enhancing Flavan-3-ol Biosynthesis in <i>Saccharomyces cerevisiae</i>

Sun, Chengcheng; Li, Guangjian; Li, Hongbiao; Lyu, Yunbin; Yu, Shiqin; Zhou, Jingwen

doi:10.1021/acs.jafc.1c04489

Cited by 21 publications

(23 citation statements)

References 45 publications

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“…There are also some studies providing a platform for the production of some industrial substances such as L-citrulline, flavan-3-ols, β-carotene, etc. by designing E. coli , lactic acid bacteria and yeast into engineered bacteria [ 62 – 66 ].…”

Section: Research and Application Of Engineered Probioticsmentioning

confidence: 99%

Engineered probiotics

Lyu

Liu

et al. 2022

Microb Cell Fact

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Engineered probiotics are a kind of new microorganisms produced by modifying original probiotics through gene editing. With the continuous development of tools and technology progresses, engineering renovation of probiotics are becoming more diverse and more feasible. In the past few years there have been some advances in the development of engineered probiotics that will benefit humankind. This review briefly introduces the theoretical basis of gene editing technology and focuses on some recent engineered probiotics researches, including inflammatory bowel disease, bacterial infection, tumor and metabolic diseases. It is hoped that it can provide help for the further development of genetically modified microorganisms, stimulate the potential of engineered probiotics to treat intractable diseases, and provide new ideas for the diagnosis of some diseases or some industrial production.

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Section: Research and Application Of Engineered Probioticsmentioning

confidence: 99%

Engineered probiotics

Lyu

Liu

et al. 2022

Microb Cell Fact

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“…The sample was then centrifuged at 9600 g for 2 min and membrane filtered (0.22 μm). Both (2 S )-naringenin and naringenin-7-O-glucoside were further characterized using liquid chromatography–mass spectrometry . A Waters MALDI SYNAPT Q-TOF MS system (Waters, MA, USA) was equipped with a BEH C18 column (2.1 mm × 100 mm, 1.7 μm; waters, MA, USA) at 40 °C.…”

Section: Methodsmentioning

confidence: 99%

“…Both (2S)-naringenin and naringenin-7-O-glucoside were further characterized using liquid chromatography−mass spectrometry. 20 A Waters MALDI SYNAPT Q-TOF MS system (Waters, MA, USA) was equipped with a BEH C18 column (2.1 mm × 100 mm, 1.7 μm; waters, MA, USA) at 40 °C. Mobile phases were: solution A (0.1% trifluoroacetic acid in acetonitrile) and solution B (0.1% trifluoroacetic acid in water) The applied A/B gradient was: 0−8 min, 5%−15% A; 8−16 min, 21% A; 16−24 min, 60% A; 24−28 min, 90% A; and 28−30 min, 5% A.…”

Section: Methodsmentioning

confidence: 99%

Glycosylation Modification Enhances (2S)-Naringenin Production in Saccharomyces cerevisiae

Lyv

et al. 2022

ACS Synth. Biol.

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(2S)-Naringenin is an important flavonoid precursor, with multiple nutritional and pharmacological activities. Both (2S)-naringenin and other flavonoid production are hindered by poor water solubility and inhibited cell growth. To address this, we increased solubility and improved cell growth by partially glycosylating (2S)-naringenin to naringenin-7-O-glucoside, which facilitated increased extracellular secretion, by knocking out endogenous glycosyl hydrolase genes, EXG1 and SPR1, and expressing the glycosyltransferase gene (UGT733C6). Naringenin-7-O-glucoside synthesis was further improved by optimizing UDP-glucose and shikimate pathways. Then, hydrochloric acid was used to hydrolyze naringenin-7-O-glucoside to (2S)-naringenin outside the cell. Thus, our optimized Saccharomyces cerevisiae strain E32T19 produced 1184.1 mg/L (2S)-naringenin, a 7.9-fold increase on the starting strain. Therefore. we propose that glycosylation modification is a useful strategy for the efficient heterologous biosynthesis of (2S)-naringenin in S. cerevisiae.

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“…7-DHC was separated in 100% methanol (mobile phase) at 280 nm at 1 mL/min in a separation time of 20 min ( Xia et al, 2022 ). LC-MS method was used to identify 7-DHC ( Sun et al, 2021 ).…”

Section: Methodsmentioning

confidence: 99%

Reengineering of 7-dehydrocholesterol biosynthesis in Saccharomyces cerevisiae using combined pathway and organelle strategies

et al. 2022

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7-Dehydrocholesterol (7-DHC) is a widely used sterol and a precursor of several costly steroidal drugs. In this study, 7-DHC biosynthesis pathway was constructed and modified in Saccharomyces cerevisiae. Firstly, the biosynthesis pathway was constructed by knocking out the competitive pathway genes ERG5 and ERG6 and integrating two DHCR24 copies from Gallus gallus at both sites. Then, 7-DHC titer was improved by knocking out MOT3, which encoded a transcriptional repressor for the 7-DHC biosynthesis pathway. Next, by knocking out NEM1 and PAH1, 7-DHC accumulation was improved, and genes upregulation was verified by quantitative PCR (qPCR). Additionally, tHMG1, IDI1, ERG2, ERG3, DHCR24, POS5, and CTT1 integration into multi-copy sites was used to convert precursors to 7-DHC, and increase metabolic flux. Finally, qPCR confirmed the significant up-regulation of key genes transcriptional levels. In a 96 h shaker flask fermentation, the 7-DHC titer was 649.5 mg/L by de novo synthesis. In a 5 L bioreactor, the 7-DHC titer was 2.0 g/L, which was the highest 7-DHC titer reported to date. Our study is of great significance for the industrial production of 7-DHC and steroid development for medical settings.

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Enhancing Flavan-3-ol Biosynthesis in Saccharomyces cerevisiae

Cited by 21 publications

References 45 publications

Engineered probiotics

Engineered probiotics

Glycosylation Modification Enhances (2S)-Naringenin Production in Saccharomyces cerevisiae

Reengineering of 7-dehydrocholesterol biosynthesis in Saccharomyces cerevisiae using combined pathway and organelle strategies

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