Efficient production of 2,3-butanediol by recombinant Saccharomyces cerevisiae through modulation of gene expression by cocktail δ-integration

Yamada, R.; Wakita, Kazuki; Mitsui, Ryosuke; Nishikawa, Riru; Ogino, Hiroyasu

doi:10.1016/j.biortech.2017.05.034

Cited by 28 publications

(18 citation statements)

References 44 publications

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“…Initially, DNA fragments encoding 15 promoters from S. cerevisiae , P. pastoris , and H. polymorpha , were PCR‐amplified using the plasmid library pPPE_LibBTLAnc (Yamada, Kimoto, & Ogino, 2016) as the template, as well as ProF and ProR as primers. All promoters we selected were used for GMES in previous studies (Yamada et al, 2017a; Yamada et al, 2017b; Yamada et al, 2017c; Yamada et al, 2018). Next, DNA fragments encoding eight enzymes ( ERG10 , ERG13 , tHMG1 , ERG12 , ERG8 , ERG19 , IDI1 , and ERG20 ) in the mevalonate pathway and their terminator sequences were PCR‐amplified using S. cerevisiae genomic DNA as the template and corresponding primers.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, the global metabolic engineering strategy (GMES) using the cocktail δ‐integration for yeast has been developed (Yamada, Wakita, & Ogino, 2017a), which involves the simultaneous integration of various multicopy genes via δ‐integration, followed by the selection of desirable transformants in S. cerevisiae . With regard to utilizing the GMES, glycolysis‐related genes (Yamada et al, 2017a), lactate production‐involved genes (Yamada, Wakita, Mitsui, & Ogino, 2017b), 2,3‐butanediol production (Yamada, Wakita, Mitsui, & Ogino, 2017c), and carotenoid production (Yamada, Yamauchi, Ando, Kumata, & Ogino, 2018) in S. cerevisiae were optimized. The GMES is a strategy that comprehensively and randomly modifies the transcription level of genes in target metabolic pathways in yeast.…”

Section: Introductionmentioning

confidence: 99%

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Construction of yeast producing patchoulol by global metabolic engineering strategy

Mitsui

Nishikawa

Yamada

et al. 2020

Biotech & Bioengineering

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Patchoulol is a sesquiterpene alcohol found in the leaves of the patchouli plant that can be extracted by steam distillation. Notably, patchoulol is an essential natural product frequently used in the chemical industry. However, patchouli produces an insignificant amount of patchoulol, not to mention steam distillation, and requires a lot of energy and time. Recombinant microorganisms that can be cultured in mild conditions and can produce patchoulol from renewable biomass resources may be a promising alternative. We previously developed the global metabolic engineering strategy (GMES), which produces a comprehensive metabolic modification in yeast, using the cocktail δ‐integration method. In this study, we aimed to produce patchoulol by modifying engineered yeast. The expression of nine genes involved in patchoulol synthesis was modulated using GMES. Regarding patchoulol production, the resultant strain, YPH499/PAT167/MVA442, showed a concentration of 42.1 mg/L, a production rate of 8.42 mg/L/d, and a yield of 2.05 mg/g‐glucose, respectably. These concentration values, production rate, and yield obtained through batch‐fermentation in this study were high level when compared to previously reported recombinant microorganism studies. GMES could be used as a potential strategy for producing secondary metabolites from plants in recombinant Saccharomyces cerevisiae.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Construction of yeast producing patchoulol by global metabolic engineering strategy

Mitsui

Nishikawa

Yamada

et al. 2020

Biotech & Bioengineering

Self Cite

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“…In yeast, delta sites are located in the transposon Ty of the yeast genome and there are over 400 delta sites in S. cerevisiae ( Lee and Da Silva, 1997 ). Genomic integration based on delta sites has been applied to the production of commercially valuable compounds by engineered S. cerevisiae , including resveratrol ( Liu et al, 2017 ), glucaric acid ( Chen et al, 2018 ), 2,3-butanediol ( Yamada et al, 2017 ), and carotenoids ( Yamada et al, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

Developing Multi-Copy Chromosomal Integration Strategies for Heterologous Biosynthesis of Caffeic Acid in Saccharomyces cerevisiae

et al. 2022

Front. Microbiol.

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Caffeic acid, a plant-sourced phenolic compound, has a variety of biological activities, such as antioxidant and antimicrobial properties. The caffeic acid biosynthetic pathway was initially constructed in S. cerevisiae, using codon-optimized TAL (coTAL, encoding tyrosine ammonia lyase) from Rhodobacter capsulatus, coC3H (encoding p-coumaric acid 3-hydroxylase) and coCPR1 (encoding cytochrome P450 reductase 1) from Arabidopsis thaliana in 2 μ multi-copy plasmids to produce caffeic acid from glucose. Then, integrated expression of coTAL via delta integration with the POT1 gene (encoding triose phosphate isomerase) as selection marker and episomal expression of coC3H, coCPR1 using the episomal plasmid pLC-c3 were combined, and caffeic acid production was proved to be improved. Next, the delta and rDNA multi-copy integration methods were applied to integrate the genes coC3H and coCPR1 into the chromosome of high p-coumaric acid yielding strain QT3-20. The strain D9 constructed via delta integration outperformed the other strains, leading to 50-fold increased caffeic acid production in optimized rich media compared with the initial construct. The intercomparison between three alternative multi-copy strategies for de novo synthesis of caffeic acid in S. cerevisiae suggested that delta-integration was effective in improving caffeic acid productivity, providing a promising strategy for the production of valuable bio-based chemicals in recombinant S. cerevisiae.

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“…Containing two chiral carbon atoms and existing in three isomer forms (R,R-, S,S-and meso-), 2,3-butanediol (2,3-BD) is a promising four-carbon biorefinery platform chemical (Celi nska and Grajek, 2009;Yan et al 2009). It is used in high-quality aviation fuel, food additives, plastics and drugs (Yan et al 2009;Ji et al 2011;Xie et al 2015;Bialkowska, 2016;Yamada et al 2017). In addition, R,R-2,3-BD is an important intermediate widely used in Letters in Applied Microbiology 69, 424--430 © 2019 The Society for Applied Microbiology the production of high-value pharmaceuticals, agricultural compounds and liquid crystals due to its unique optical activity (Ji et al 2015;Bialkowska, 2016;Yamada et al 2017).…”

Section: Introductionmentioning

confidence: 99%

Effects of amino acids on the fermentation of inulin or glucose to produceR,R‐2,3‐butanediol usingPaenibacillus polymyxaZJ‐9

Gao

Jiang

Chun-xiao

et al. 2019

Lett Appl Microbiol

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This study investigated the effects of different amino acids on the fermentation of pure inulin and glucose using Paenibacillus polymyxa ZJ‐9 in order to improve the production of R,R‐2,3‐butanediol (R,R‐2,3‐BD) respectively. The inulin extract from Jerusalem artichoke tubers contained 19 common amino acids, which were detected by HPLC. Arg featured the highest content (1290 mg l−1). A single add‐back experiment of 20 common amino acids indicated that Asn, Ser, His and Arg are key amino acids in R,R‐2,3‐BD synthesis during inulin fermentation using P. polymyxa ZJ‐9. The corresponding yields of R,R‐2,3‐BD reached 24·32, 22·32, 22·03 and 21·04 g l−1 after the four key amino acids (1·5 g l−1 each) and glucose were evaluated in this fermentation. The yields were considerably higher than that of the control group (12·11 g l−1). With the addition of the mixture of four amino acids (1·5 g l−1 each), the highest yields of R,R‐2,3‐BD (25·07 and 17·47 g l−1) were obtained with the increase of 107·0 and 89·1% during the fermentation of glucose and pure inulin respectively. Significance and Impact of the Study Paenibacillus polymyxa is a micro‐organism with a reported potential for industrialized production of R,R‐2,3‐butanediol. The nitrogen sources have a significant effect on R,R‐2,3‐butanediol formation using P. polymyxa. This study demonstrated a highly efficient new method to improve the yield of R,R‐2,3‐butanediol without adding other nitrogen sources except amino acids during the fermentation. This will therefore decrease the production cost of R,R‐2,3‐butanediol and provide a new strategy for promoting synthesis of amino acid‐dependent products.

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Efficient production of 2,3-butanediol by recombinant Saccharomyces cerevisiae through modulation of gene expression by cocktail δ-integration

Cited by 28 publications

References 44 publications

Construction of yeast producing patchoulol by global metabolic engineering strategy

Construction of yeast producing patchoulol by global metabolic engineering strategy

Developing Multi-Copy Chromosomal Integration Strategies for Heterologous Biosynthesis of Caffeic Acid in Saccharomyces cerevisiae

Effects of amino acids on the fermentation of inulin or glucose to produceR,R‐2,3‐butanediol usingPaenibacillus polymyxaZJ‐9

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