Modular Engineering of Tyrosol Production in <i>Escherichia coli</i>

Yang, Haiquan; Xue, Yuxiang; Yang, Chen; Shen, Wei; Fan, You; Chen, Xianzhong

doi:10.1021/acs.jafc.9b00227

Cited by 21 publications

(14 citation statements)

References 26 publications

(74 reference statements)

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“…Tyrosol, which is a valuable chemical for the synthesis of pharmaceuticals, was produced at 2.8 g/liter from 10 g/liter glucose with a yield of 0.28 g/g glucose (0.36 mol/mol glucose). The yield was much higher than that observed in previous studies (53)(54)(55). Other aromatic compounds have been successfully produced using engineered E. coli, e.g., phenol (35), DOPA (56, 57), salvianic acid A (58), caffeyl and coniferyl alcohol (59), 4-hydroxymandelic acid (60), and phenylacetic acid and 4-hydroxyphenylacetic acid (36).…”

Section: Discussionmentioning

confidence: 87%

Chromosome Engineering To Generate Plasmid-Free Phenylalanine- and Tyrosine-Overproducing Escherichia coli Strains That Can Be Applied in the Generation of Aromatic-Compound-Producing Bacteria

Koma

Kishida

Yoshida

et al. 2020

Appl Environ Microbiol

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ABSTRACT Many phenylalanine- and tyrosine-producing strains have used plasmid-based overexpression of pathway genes. The resulting strains achieved high titers and yields of phenylalanine and tyrosine. Chromosomally engineered, plasmid-free producers have shown lower titers and yields than plasmid-based strains, but the former are advantageous in terms of cultivation cost and public health/environmental risk. Therefore, we engineered here the Escherichia coli chromosome to create superior phenylalanine- and tyrosine-overproducing strains that did not depend on plasmid-based expression. Integration into the E. coli chromosome of two central metabolic pathway genes (ppsA and tktA) and eight shikimate pathway genes (aroA, aroB, aroC, aroD, aroE, aroGfbr, aroL, and pheAfbr), controlled by the T7lac promoter, resulted in excellent titers and yields of phenylalanine; the superscript “fbr” indicates that the enzyme encoded by the gene was feedback resistant. The generated strain could be changed to be a superior tyrosine-producing strain by replacing pheAfbr with tyrAfbr. A rational approach revealed that integration of seven genes (ppsA, tktA, aroA, aroB, aroC, aroGfbr, and pheAfbr) was necessary as the minimum gene set for high-yield phenylalanine production in E. coli MG1655 (tyrR, adhE, ldhA, pykF, pflDC, and ascF deletant). The phenylalanine- and tyrosine-producing strains were further applied to generate phenyllactic acid-, 4-hydroxyphenyllactic acid-, tyramine-, and tyrosol-producing strains; yield of these aromatic compounds increased proportionally to the increase in phenylalanine and tyrosine yields. IMPORTANCE Plasmid-free strains for aromatic compound production are desired in the aspect of industrial application. However, the yields of phenylalanine and tyrosine have been considerably lower in plasmid-free strains than in plasmid-based strains. The significance of this research is that we succeeded in generating superior plasmid-free phenylalanine- and tyrosine-producing strains by engineering the E. coli chromosome, which was comparable to that in plasmid-based strains. The generated strains have a potential to generate superior strains for the production of aromatic compounds. Actually, we demonstrated that four kinds of aromatic compounds could be produced from glucose with high yields (e.g., 0.28 g tyrosol/g glucose).

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Section: Discussionmentioning

confidence: 87%

Chromosome Engineering To Generate Plasmid-Free Phenylalanine- and Tyrosine-Overproducing Escherichia coli Strains That Can Be Applied in the Generation of Aromatic-Compound-Producing Bacteria

Koma

Kishida

Yoshida

et al. 2020

Appl Environ Microbiol

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“…Conversely, microbial biosynthesis provides a sustainable and economically feasible platform for producing natural products originally derived from plants (Lee et al ., 2012 ; Borodina and Nielsen, 2014 ; Paddon and Keasling, 2014 ). In recent years, E. coil was successfully used to produce tyrosol and salidroside (Satoh et al ., 2012 ; Bai et al ., 2014 ; Chung et al ., 2017 ; Fan et al ., 2017 ; Xue et al ., 2017 ; Liu et al ., 2018 ; Yang et al ., 2019 ). Through these studies, titres of tyrosol achieved using sugar (glucose, or xylose and glucose mixture) as the carbon source, have increased from 69.08 mg l −1 (0.5 mM) to 1.47 g l −1 (Satoh et al ., 2012 ; Liu et al ., 2018 ; Yang et al ., 2019 ).…”

Section: Introductionmentioning

confidence: 99%

“…In recent years, E. coil was successfully used to produce tyrosol and salidroside (Satoh et al ., 2012 ; Bai et al ., 2014 ; Chung et al ., 2017 ; Fan et al ., 2017 ; Xue et al ., 2017 ; Liu et al ., 2018 ; Yang et al ., 2019 ). Through these studies, titres of tyrosol achieved using sugar (glucose, or xylose and glucose mixture) as the carbon source, have increased from 69.08 mg l −1 (0.5 mM) to 1.47 g l −1 (Satoh et al ., 2012 ; Liu et al ., 2018 ; Yang et al ., 2019 ). The production of salidroside from sugar increased from 56.90 to 670.58 mg l −1 in shake flasks and to 6.03 g l −1 in a 5 l bioreactor, using a codon‐optimized UDP‐glycosyltransferase synUgt85a1 from Arabidopsis thaliana and adopting a co‐culture strategy (Liu et al ., 2018 ).…”

Section: Introductionmentioning

confidence: 99%

Multi‐modular engineering of Saccharomyces cerevisiae for high‐titre production of tyrosol and salidroside

Liu

Tian

Zhou

et al. 2020

Microbial Biotechnology

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Tyrosol and its glycosylated product salidroside are important ingredients in pharmaceuticals, nutraceuticals and cosmetics. Despite the ability of Saccharomyces cerevisiae to naturally synthesize tyrosol, high yield from de novo synthesis remains a challenge. Here, we used metabolic engineering strategies to construct S. cerevisiae strains for high-level production of tyrosol and salidroside from glucose. First, tyrosol production was unlocked from feedback inhibition. Then, transketolase and ribose-5phosphate ketol-isomerase were overexpressed to balance the supply of precursors. Next, chorismate synthase and chorismate mutase were overexpressed to maximize the aromatic amino acid flux towards tyrosol synthesis. Finally, the competing pathway was knocked out to further direct the carbon flux into tyrosol synthesis. Through a combination of these interventions, tyrosol titres reached 702.30 AE 0.41 mg l À1 in shake flasks, which were approximately 26-fold greater than that of the WT strain. RrU8GT33 from Rhodiola rosea was also applied to cells and maximized salidroside production from tyrosol in S. cerevisiae. Salidroside titres of 1575.45 AE 19.35 mg l À1 were accomplished in shake flasks. Furthermore, titres of 9.90 AE 0.06 g l À1 of tyrosol and 26.55 AE 0.43 g l À1 of salidroside were achieved in 5 l bioreactors, both are the highest titres reported to date. The synergistic engineering strategies presented in this study could be further applied to increase the production of high valueadded aromatic compounds derived from the aromatic amino acid biosynthesis pathway in S. cerevisiae.

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“…S1/S9a). The theoretical copy numbers of plasmids pACYCDuet-1, pCDFDuet-1, pETDuet-1, and pRSFDuet-1 are 10, 20, 40, and 100, respectively [ 36 ]. In this study, the actual copy numbers of empty plasmids pACYCDuet-1, pCDFDuet-1, pETDuet-1, and pRSFDuet-1 are 3.38, 4.89, 9.12, and 26.3 at the cultivation conditions, respectively (Additional file 1 : Fig.…”

Section: Resultsmentioning

confidence: 99%

Efficient production of cembratriene-ol in Escherichia coli via systematic optimization

et al. 2023

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Background The tobacco leaf–derived cembratriene-ol exhibits anti-insect effects, but its content in plants is scarce. Cembratriene-ol is difficult and inefficiently chemically synthesised due to its complex structure. Moreover, the titer of reported recombinant hosts producing cembratriene-ol was low and cannot be applied to industrial production. Results In this study, Pantoea ananatis geranylgeranyl diphosphate synthase (CrtE) and Nicotiana tabacum cembratriene-ol synthase (CBTS) were heterologously expressed to synthsize the cembratriene-ol in Escherichia coli. Overexpression of cbts*, the 1-deoxy-d-xylulose 5-phosphate synthase gene dxs, and isopentenyl diphosphate isomerase gene idi promoted the production of cembratriene-ol. The cembratriene-ol titer was 1.53-folds higher than that of E. coli Z17 due to the systematic regulation of ggpps, cbts*, dxs, and idi expression. The production of cembratriene-ol was boosted via the overexpression of genes ispA, ispD, and ispF. The production level of cembratriene-ol in the optimal medium at 72 h was 8.55-folds higher than that before fermentation optimisation. The cembratriene-ol titer in the 15-L fermenter reached 371.2 mg L− 1, which was the highest titer reported. Conclusion In this study, the production of cembratriene-ol in E. coli was significantly enhanced via systematic optimization. It was suggested that the recombinant E. coli producing cembratriene-ol constructed in this study has potential for industrial production and applications.

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Modular Engineering of Tyrosol Production in Escherichia coli

Cited by 21 publications

References 26 publications

Chromosome Engineering To Generate Plasmid-Free Phenylalanine- and Tyrosine-Overproducing Escherichia coli Strains That Can Be Applied in the Generation of Aromatic-Compound-Producing Bacteria

Chromosome Engineering To Generate Plasmid-Free Phenylalanine- and Tyrosine-Overproducing Escherichia coli Strains That Can Be Applied in the Generation of Aromatic-Compound-Producing Bacteria

Multi‐modular engineering of Saccharomyces cerevisiae for high‐titre production of tyrosol and salidroside

Efficient production of cembratriene-ol in Escherichia coli via systematic optimization

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