Comparative metabolic profiling of engineered Saccharomyces cerevisiae with enhanced flavonoids production

Lyu, Xiaomei; Ng, Kuan Rei; Mark, Rita; Lee, Jie Lin; Chen, Wei Ning

doi:10.1016/j.jff.2018.03.012

Cited by 9 publications

(7 citation statements)

References 32 publications

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“…ARO4 K229L and ARO7 G141S are the mutated feedback-resistant enzymes of the shikimate pathway. They are capable of eliminating the feedback inhibition of DAHP (3-deoxy- d -arabino-heptulosonate 7-phosphate) and increase the level of NADPH, which is vital to the biosynthesis of flavonoids in S. cerevisiae.…”

Section: Resultsmentioning

confidence: 99%

“…ARO4 K229L and ARO7 G141S are the mutated feedback-resistant enzymes of the shikimate pathway. They are capable of eliminating the feedback inhibition of DAHP (3-deoxy-D-arabino-heptulosonate 7-phosphate) and increase the level of NADPH, 30 which is vital to the biosynthesis of flavonoids in S. cerevisiae. PDC5 (pyruvate decarboxylase) and ARO10 (phenylpyruvate decarboxylase) are active phenylpyruvate decarboxylases; by knocking out both of them, the amount of byproducts, such as phenyl ethanol, ρ-hydroxyphenyl ethanol, and related acids, would be decreased drastically.…”

Section: Resultsmentioning

confidence: 99%

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Enhanced Biosynthesis of Dihydromyricetin in Saccharomyces cerevisiae by Coexpression of Multiple Hydroxylases

Lyu

et al. 2020

J. Agric. Food Chem.

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Dihydromyricetin (DHM) is a traditional plant-extracted flavonoid with some health benefits. This study aimed to metabolically engineer the strains for DHM bioproduction. Two strains of BK-11 and BQ-21 were integrated with flavonoid 3hydroxylase (F3H) or both F3H and flavonoid 3′-hydroxylase (F3′H). The resulting strains have expressed the enzymes of GmCPR and SlF3′5′H, and then, the promoters of INO1p and TDH1p were used to enhance further the DHM production from naringenin in Saccharomyces cerevisiae. Through multiple-copy integration, 709.6 mg/L DHM was obtained by adding 2.5 g/L naringenin in a 5 L bioreactor, implying that the synergistic effect between F3′H and flavonoid 3′5′-hydroxylase is likely to promote the DHM production. An yield of 246.4 mg/L DHM was obtained from glucose by deleting genes for branch pathways and integrating PhCHS, MsCHI, Pc4CL, and FjTAL. To our knowledge, this is the highest production reported for the de novo biosynthesis of DHM.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Enhanced Biosynthesis of Dihydromyricetin in Saccharomyces cerevisiae by Coexpression of Multiple Hydroxylases

Lyu

et al. 2020

J. Agric. Food Chem.

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“…In addition, gene and protein expression levels are also changed dramatically in the metabolic process of plants under such conditions. The application of metabolomics methods offers a rational way to reveal the implications of these metabolic changes on a broad scale [31]. Currently, metabolomics-based technologies have been broadly employed to clarify the metabolic responses of various plants (both halophytes and glycophytes) to salt stress, such as Hordeum vulgare [32], Oryza sativa [33], Suaeda corniculata [34], Glycine max [30], and Zea mays [35].…”

Section: Introductionmentioning

confidence: 99%

Metabolomics Analysis Reveals the Alkali Tolerance Mechanism in Puccinellia tenuiflora Plants Inoculated with Arbuscular Mycorrhizal Fungi

et al. 2020

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Soil alkalization is a major environmental threat that affects plant distribution and yield in northeastern China. Puccinellia tenuiflora is an alkali-tolerant grass species that is used for salt-alkali grassland restoration. However, little is known about the molecular mechanisms by which arbuscular mycorrhizal fungi (AMF) enhance P. tenuiflora responses to alkali stress. Here, metabolite profiling in P. tenuiflora seedlings with or without arbuscular mycorrhizal fungi (AMF) under alkali stress was conducted using liquid chromatography combined with time-of-flight mass spectrometry (LC/TOF-MS). The results showed that AMF colonization increased seedling biomass under alkali stress. In addition, principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA) demonstrated that non-AM and AM seedlings showed different responses under alkali stress. A heat map analysis showed that the levels of 88 metabolites were significantly changed in non-AM seedlings, but those of only 31 metabolites were significantly changed in AM seedlings. Moreover, the levels of a total of 62 metabolites were significantly changed in P. tenuiflora seedlings after AMF inoculation. The results suggested that AMF inoculation significantly increased amino acid, organic acid, flavonoid and sterol contents to improve osmotic adjustment and maintain cell membrane stability under alkali stress. P. tenuiflora seedlings after AMF inoculation produced more plant hormones (salicylic acid and abscisic acid) than the non-AM seedlings, probably to enhance the antioxidant system and facilitate ion balance under stress conditions. In conclusion, these findings provide new insights into the metabolic mechanisms of P. tenuiflora seedlings with arbuscular mycorrhizal fungi under alkali conditions and clarify the role of AM in the molecular regulation of this species under alkali stress.

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“…The plasmid used for the experiment is pCEV-G1-Km, where 4CL and CHS were cloned and expressed. While the Y-28 strains contained the pCEV-G1-Ph plasmid expressing TAL from Flavobacterium johnsoniae and CHI from Medicago sativa, which were given by Lyu et al…”

Section: Methodsmentioning

confidence: 99%

“…The plasmid used for the experiment is pCEV-G1-Km, 26 where 4CL and CHS were cloned and expressed. While the Y-28 strains contained the pCEV-G1-Ph plasmid expressing TAL from Flavobacterium johnsoniae and CHI from Medicago sativa, which were given by Lyu et al 28 All 4CL and CHS genes used in this study were synthesized by GenScipt, with codon optimization according to S. cerevisiae and cloned in pUC57. The DNA sequences were obtained from the NCBI GenBank with the following reference numbers for 4CL: A. thaliana (NM_113019.3), M. truncatula (XM_003610795.2), P. crispum (KF765780.1), and S. coelicolor (NC_003888.3) and for CHS: A. thaliana (NM_121396.3), G. uralensis (EF026979.1), H. androsaemum (AF315345.1), and V. vinifera (NM_001280950.1).…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

Gene Source Screening as a Tool for Naringenin Production in Engineered Saccharomyces cerevisiae

Mark

Lyu

et al. 2019

ACS Omega

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Flavonoids are plant secondary metabolites with great potential in the food industry. Metabolic engineering of Saccharomyces cerevisiae is a sustainable production technique. However, the current naringenin production yield is low because of inefficient enzymatic activity. Hence, this study uses gene source screening as a tool to identify the best gene source for enzymes such as 4-coumarate: coenzyme ligase (4CL) and chalcone synthase (CHS). For the first time, the 4CL gene from Medicago truncatula and the CHS gene from Vitis vinifera were expressed in S. cerevisiae, and this combination provided the highest yield of naringenin, which was 28-fold higher as compared to the reference strain. The combinations obtained similar performance in the Y-28 strains, where the highest production was 28.68 mg/L. Our results demonstrated that the selection and combination of enzymes from the correct gene source could greatly improve naringenin production. For the future, this could help commercialize flavonoid production, which would result in natural food preservatives and additives.

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Comparative metabolic profiling of engineered Saccharomyces cerevisiae with enhanced flavonoids production

Cited by 9 publications

References 32 publications

Enhanced Biosynthesis of Dihydromyricetin in Saccharomyces cerevisiae by Coexpression of Multiple Hydroxylases

Enhanced Biosynthesis of Dihydromyricetin in Saccharomyces cerevisiae by Coexpression of Multiple Hydroxylases

Metabolomics Analysis Reveals the Alkali Tolerance Mechanism in Puccinellia tenuiflora Plants Inoculated with Arbuscular Mycorrhizal Fungi

Gene Source Screening as a Tool for Naringenin Production in Engineered Saccharomyces cerevisiae

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