Rational Engineering of Chorismate-Related Pathways in Saccharomyces cerevisiae for Improving Tyrosol Production

Guo, Wei; Huang, Qiulan; Líu, Hao; Hou, Shaoli; Niu, Suhao; Jiang, Yi; Bao, Xinhe; Shen, Yu; Xu, Fang

doi:10.3389/fbioe.2019.00152

Cited by 30 publications

(20 citation statements)

References 42 publications

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“…Salidroside is produced via the glycosylation of tyrosol at the 8‐OH group, which is catalysed by a regio‐specific uridine 5’‐diphospho‐glucosyltransferase (UGT; Fan et al ., 2017 ). Previous studies increased tyrosol titres to 927.68 ± 25.26 mg l −1 in shake flasks and 8.37 g l −1 in a bioreactor in a haploid industrial S. cerevisiae strain HLF‐Dα (Jiang et al ., 2018 ; Torrens‐Spence et al ., 2018 ; Guo et al ., 2019 ; Guo et al ., 2020 ). The highest salidroside titres reported for engineered S. cerevisiae are 239.5 mg l −1 in a shake flask and 1.82 g l −1 in a bioreactor (Jiang et al ., 2018 ; Torrens‐Spence et al ., 2018 ; Guo et al ., 2020 ).…”

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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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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“…βglucosidase activity was measured as we previously described, except that the temperature was 30 o C. 36 The unit of β-glucosidase activity were defined as the amount of enzyme The products were analyzed by HPLC as we previously described. 37 Statistics. The Student's t-test was performed for significant differences between two groups of data.…”

Section: Discussionmentioning

confidence: 99%

“…The products were analyzed by HPLC as we previously described. 37 An Inertsil NH2 column (250 mm × 7.8 mm; Shimadzu, Kyoto, Japan) and 80% acetonitrile as the mobile phase (1.0 ml/min, 45 °C) were used. Synthesized disaccharides products were detected by TLC following the removal of glucose from the solution.…”

Section: Introductionmentioning

confidence: 99%

Efficient Synthesis of Rare Disaccharides by Engineered β-Glucosidase Based on Hydropathy Index

Niu¹,

Liu²,

Feng³

et al. 2020

Preprint

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Oligosaccharides have important therapeutic applications. A useful route for oligosaccharides synthesis, especially rare disaccharides, is reverse hydrolysis by β-glucosidase. However, the low conversion efficiency of disaccharides from monosaccharides limits its large-scale production because the equilibrium is biased in the direction of hydrolysis. Based on the analysis of the docking results, we hypothesized that the hydropathy index of key amino acid residues in the catalytic site is closely related with disaccharide synthesis and more hydrophilic residues located in the catalytic site would enhance reverse hydrolysis activity. In this study, positive variants TrCel1bI177S, TrCel1bI177S/I174S, and TrCel1bI177S/I174S/W173H, and one negative variant TrCel1bN240I were designed according to the Hydropathy Index For Enzyme Activity (HIFEA) strategy. The reverse hydrolysis with TrCel1bI177S/I174S/W173H was accelerated and then the maximum total production (<a>195.8 mg/ml/mg enzyme</a>) of the synthesized disaccharides was increased 3.5-fold compared to that of wildtype. On the contrary, <a>TrCel1b</a>N240I lost reverse hydrolysis activity. The results demonstrate that<a> </a><a>the average hydropathy index</a> of <a>the key amino acid residues </a>in the catalytic site of TrCel1b is an important factor for the synthesis of laminaribiose, sophorose, and cellobiose. The HIFEA strategy provides a new perspective for the rational design of β-glucosidases used for the synthesis of oligosaccharides.

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“…Chromosomal overexpression of P. crispum aas, A. thaliana UGT85A1 as well as the enhancement of the expressions of ARO4 K229L , ARO7 G141S , and AROL resulted in the production of tyrosol and salidroside to 1394.6 and 732.5 mg/L, respectively (Jiang et al, 2018). An AAS-derived pathway containing P. crispum Aas and E. coli Adh was overexpressed for the production of tyrosol in S. cerevisiae (Guo et al, 2019). The replacement of PDC1 with E. coli tyr mutant tyrA M53I/A354V further improved the production of tyrosol by 440 times.…”

Section: S Cerevisiae S288cmentioning

confidence: 99%

Recent Advances in Metabolically Engineered Microorganisms for the Production of Aromatic Chemicals Derived From Aromatic Amino Acids

Shen

Niu

Yan

et al. 2020

Front. Bioeng. Biotechnol.

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Aromatic compounds derived from aromatic amino acids are an important class of diverse chemicals with a wide range of industrial and commercial applications. They are currently produced via petrochemical processes, which are not sustainable and eco-friendly. In the past decades, significant progress has been made in the construction of microbial cell factories capable of effectively converting renewable carbon sources into value-added aromatics. Here, we systematically and comprehensively review the recent advancements in metabolic engineering and synthetic biology in the microbial production of aromatic amino acid derivatives, stilbenes, and benzylisoquinoline alkaloids. The future outlook concerning the engineering of microbial cell factories for the production of aromatic compounds is also discussed.

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Rational Engineering of Chorismate-Related Pathways in Saccharomyces cerevisiae for Improving Tyrosol Production

Cited by 30 publications

References 42 publications

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

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

Efficient Synthesis of Rare Disaccharides by Engineered β-Glucosidase Based on Hydropathy Index

Recent Advances in Metabolically Engineered Microorganisms for the Production of Aromatic Chemicals Derived From Aromatic Amino Acids

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