Refactoring of a synthetic raspberry ketone pathway with EcoFlex

Hleba, Yonek B; Bischoff, Sarah; Bell, David; Polizzi, Karen M.; Freemont, Paul S.

doi:10.1186/s12934-021-01604-4

Cited by 15 publications

(16 citation statements)

References 30 publications

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“…Frontiers in Bioengineering and Biotechnology frontiersin.org raspberry ketone-producing pathway was reconstructed in E. coli (Moore et al, 2021), and led to 12.9 mg/L raspberry ketone production by fine-tuning the expression level of each pathway gene using a promoter library. Zingerone is another phenylbutanones and the key component responsible for the pungency of ginger.…”

Section: Benzalacetone Synthasementioning

confidence: 99%

Engineered biosynthesis of plant polyketides by type III polyketide synthases in microorganisms

Liu

2022

Front. Bioeng. Biotechnol.

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Plant specialized metabolites occupy unique therapeutic niches in human medicine. A large family of plant specialized metabolites, namely plant polyketides, exhibit diverse and remarkable pharmaceutical properties and thereby great biomanufacturing potential. A growing body of studies has focused on plant polyketide synthesis using plant type III polyketide synthases (PKSs), such as flavonoids, stilbenes, benzalacetones, curcuminoids, chromones, acridones, xanthones, and pyrones. Microbial expression of plant type III PKSs and related biosynthetic pathways in workhorse microorganisms, such as Saccharomyces cerevisiae, Escherichia coli, and Yarrowia lipolytica, have led to the complete biosynthesis of multiple plant polyketides, such as flavonoids and stilbenes, from simple carbohydrates using different metabolic engineering approaches. Additionally, advanced biosynthesis techniques led to the biosynthesis of novel and complex plant polyketides synthesized by diversified type III PKSs. This review will summarize efforts in the past 10 years in type III PKS-catalyzed natural product biosynthesis in microorganisms, especially the complete biosynthesis strategies and achievements.

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Section: Benzalacetone Synthasementioning

confidence: 99%

Engineered biosynthesis of plant polyketides by type III polyketide synthases in microorganisms

Liu

2022

Front. Bioeng. Biotechnol.

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“…coli . Moore et al used the toolkit EcoFlex to establish a combinatorial promoter library to optimize the expression levels of the genes in the RK synthesis pathway including TAL , PCL , BAS , RKS , and MatB . The final engineered E.…”

Section: Introductionmentioning

confidence: 99%

Modular Metabolic Engineering and Synthetic Coculture Strategies for the Production of Aromatic Compounds in Yeast

et al. 2023

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Microbial-derived aromatics provide a sustainable and renewable alternative to petroleum-derived chemicals. In this study, we used the model yeast Saccharomyces cerevisiae to produce aromatic molecules by exploiting the concept of modularity in synthetic biology. Three different modular approaches were investigated for the production of the valuable fragrance raspberry ketone (RK), found in raspberry fruits and mostly produced from petrochemicals. The first strategy used was modular cloning, which enabled the generation of combinatorial libraries of promoters to optimize the expression level of the genes involved in the synthesis pathway of RK. The second strategy was modular pathway engineering and involved the creation of four modules, one for product formation: RK synthesis module (Mod. RK); and three for precursor synthesis: aromatic amino acid synthesis module (Mod. Aro), p-coumaric acid synthesis module (Mod. p-CA), and malonyl-CoA synthesis module (Mod. M-CoA). The production of RK by combinations of the expression of these modules was studied, and the best engineered strain produced 63.5 mg/L RK from glucose, which is the highest production described in yeast, and 2.1 mg RK/g glucose, which is the highest yield reported in any organism without p-coumaric acid supplementation. The third strategy was the use of modular cocultures to explore the effects of division of labor on RK production. Two two-member communities and one three-member community were created, and their production capacity was highly dependent on the structure of the synthetic community, the inoculation ratio, and the culture media. In certain conditions, the cocultures outperformed their monoculture controls for RK production, although this was not the norm. Interestingly, the cocultures showed up to 7.5-fold increase and 308.4 mg/L of 4-hydroxy benzalacetone, the direct precursor of RK, which can be used for the semi-synthesis of RK. This study illustrates the utility of modularity in synthetic biology tools and their applications to the synthesis of products of industrial interest.

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“…Phenylalanine and tyrosine can be de‐aminated by phenylalanine/tyrosine ammonia lyase (PAL/TAL) to produce cinnamic acid and p ‐coumaric acid, respectively, from which other phenylpropanoids are derived. However, production can be limited by the toxicity of phenylpropanoids towards the production hosts (Lou et al, 2012; Vestergaard & Ingmer, 2019), as well as flux limitations through certain key reactions (Gu et al, 2020; Moore et al, 2021). Genetic engineering strategies have been used to improve tolerance of production hosts towards phenylpropanoids, improve flux through rate‐limiting steps, and increase the supply of metabolic precursors and co‐factors.…”

Section: Introductionmentioning

confidence: 99%

Combining genetic engineering and bioprocess concepts for improved phenylpropanoid production

Virklund¹,

Jensen

Nielsen

et al. 2022

Biotech & Bioengineering

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The group of natural aromatic compounds known as phenylpropanoids has diverse applications, but current methods of production which are largely based on synthesis from petrochemicals or extraction from agricultural biomass are unsustainable. Bioprocessing is a promising alternative, but improvements in production titers and rates are required to make this method profitable. Here the recent advances in genetic engineering and bioprocess concepts for the production of phenylpropanoids are presented for the purpose of identifying successful strategies, including adaptive laboratory evolution, enzyme engineering, in‐situ product removal, and biocatalysis. The pros and cons of bacterial and yeast hosts for phenylpropanoid production are discussed, also in the context of different phenylpropanoid targets and bioprocess concepts. Finally, some broad recommendations are made regarding targets for continued improvement and areas requiring specific attention from researchers to further improve production titers and rates.

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Refactoring of a synthetic raspberry ketone pathway with EcoFlex

Cited by 15 publications

References 30 publications

Engineered biosynthesis of plant polyketides by type III polyketide synthases in microorganisms

Engineered biosynthesis of plant polyketides by type III polyketide synthases in microorganisms

Modular Metabolic Engineering and Synthetic Coculture Strategies for the Production of Aromatic Compounds in Yeast

Combining genetic engineering and bioprocess concepts for improved phenylpropanoid production

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