Rapid prototyping of microbial production strains for the biomanufacture of potential materials monomers

Robinson, Christopher J.; Carbonell, Pablo; Jervis, Adrian J.; Yan, Cunyu; Hollywood, Katherine A.; Dunstan, Mark S.; Currin, Andrew; Swainston, Neil; Spiess, Reynard; Taylor, Sandra; Mulherin, Paul; Parker, Steven J.; Rowe, William; Matthews, Nicholas E.; Malone, Kirk J.; Feuvre, Rosalind Le; Shapira, Philip; Barran, Perdita E.; Turner, Nicholas J.; Micklefield, Jason; Breitling, Rainer; Takano, Eriko; Scrutton, Nigel S.

doi:10.1016/j.ymben.2020.04.008

Cited by 57 publications

(86 citation statements)

References 67 publications

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“…To see if we could achieve similar success for naringenin, we switched the PAL and 4CL genes with the three TAL and five 4CL candidates to give a total of 15 pathway variants ( Figure 1B ). Due to the small library size, the full combinatorial library of expression constructs was assembled using our automated pathway assembly pipeline ( 8 , 9 , 27–29 ) ( Supplementary Figure 1 ). In brief, gene parts were designed to be compatible with DNA assembly via the ligase cycling reaction, and in silico generated pipetting worklists were generated to drive the liquid handling robotic platforms.…”

Section: Resultsmentioning

confidence: 99%

“…Having established the optimal genetic pathway construct and best fermentation conditions, we looked to streamline our fermentation conditions by removing additional feedstock requirements. We previously constructed a gene double knockout (DKO) strain to boost the metabolic flux towards tyrosine overproduction (Δ tyrR , Δ pheLA ) ( 9 ). We deleted the tyrR gene, a transcriptional regulator of aromatic amino acid biosynthesis, from the DH5α genome and then deleted pheLA (bifunctional chorismate mutase/prephenate dehydratase) to boost tyrosine levels and restrict phenylalanine production ( Supplementary Figure S7 ).…”

Section: Resultsmentioning

confidence: 99%

“…The pathways were built using an automated ligase cycling reaction assembly method as described previously ( 8 , 9 , 24 ). Automated, worklist-driven liquid handling was implemented for bridging oligo pooling and ligase cycling reaction setup.…”

Section: Methodsmentioning

confidence: 99%

“…Biosynthesis via microbial fermentation offers an attractive alternative, with great strides having been made over the last 10 years in engineering biological systems to produce a number of industrial targets. Engineering microbial hosts for the production of fine and specialty chemicals, including flavonoids, offers an attractive and green route to these compounds and has become an ongoing focus of the synthetic biology community ( 8 , 9 ).…”

Section: Introductionmentioning

confidence: 99%

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Engineering Escherichia coli towards de novo production of gatekeeper (2S)-flavanones: naringenin, pinocembrin, eriodictyol and homoeriodictyol

et al. 2020

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Natural plant-based flavonoids have drawn significant attention as dietary supplements due to their potential health benefits, including anti-cancer, anti-oxidant, and anti-asthmatic activities. Naringenin, pinocembrin, eriodictyol and homoeriodictyol are classified as (2S)-flavanones, an important sub-group of naturally-occurring flavonoids, with wide-reaching applications in human health and nutrition. These four compounds occupy a central position as branch point intermediates towards a broad spectrum of naturally occurring flavonoids. Here, we report the development of E. coli production chassis for each of these key gatekeeper flavonoids. Selection of key enzymes, genetic construct design, and the optimization of process conditions resulted in the highest reported titers for naringenin (484 mg/L), improved production of pinocembrin (198 mg/L) and eriodictyol (55 mg/L from caffeic acid), and provided the first example of in vivo production of homoeriodictyol directly from glycerol (17 mg/L). This work provides a springboard for future production of diverse downstream natural and non-natural flavonoid targets.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Engineering Escherichia coli towards de novo production of gatekeeper (2S)-flavanones: naringenin, pinocembrin, eriodictyol and homoeriodictyol

et al. 2020

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“…Significant progress towards designing and building the 16 synthetic chromosomes of yeast is already being achieved [32]. Recently, scientists at SYNBIOCHEM, the University of Manchester-based SBRC, have successfully demonstrated the effectiveness of the DBLT engineering biology approachautomating the biomanufacturing compound agnostic pipeline for the on-demand production of a diverse range of industrially relevant chemical building blocks [33].…”

Section: The Bioeconomy Drivermentioning

confidence: 99%

Developing synthetic biology for industrial biotechnology applications

Clarke

Kitney

2020

Biochemical Society Transactions

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Since the beginning of the 21st Century, synthetic biology has established itself as an effective technological approach to design and engineer biological systems. Whilst research and investment continues to develop the understanding, control and engineering infrastructural platforms necessary to tackle ever more challenging systems — and to increase the precision, robustness, speed and affordability of existing solutions — hundreds of start-up companies, predominantly in the US and UK, are already translating learnings and potential applications into commercially viable tools, services and products. Start-ups and SMEs have been the predominant channel for synthetic biology commercialisation to date, facilitating rapid response to changing societal interests and market pull arising from increasing awareness of health and global sustainability issues. Private investment in start-ups across the US and UK is increasing rapidly and now totals over $12bn. Health-related biotechnology applications have dominated the commercialisation of products to date, but significant opportunities for the production of bio-derived materials and chemicals, including consumer products, are now being developed. Synthetic biology start-ups developing tools and services account for between 10% (in the UK) and ∼25% (in the US) of private investment activity. Around 20% of synthetic biology start-ups address industrial biotechnology targets, but currently, only attract ∼11% private investment. Adopting a more networked approach — linking specialists, infrastructure and ongoing research to de-risk the economic challenges of scale-up and supported by an effective long-term funding strategy — is set to transform the impact of synthetic biology and industrial biotechnology in the bioeconomy.

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Biorefinery Products as Naturally‐Based Key Raw Materials for Adhesives

Fink

2023

Biobased Adhesives

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Rapid prototyping of microbial production strains for the biomanufacture of potential materials monomers

Cited by 57 publications

References 67 publications

Engineering Escherichia coli towards de novo production of gatekeeper (2S)-flavanones: naringenin, pinocembrin, eriodictyol and homoeriodictyol

Engineering Escherichia coli towards de novo production of gatekeeper (2S)-flavanones: naringenin, pinocembrin, eriodictyol and homoeriodictyol

Developing synthetic biology for industrial biotechnology applications

Biorefinery Products as Naturally‐Based Key Raw Materials for Adhesives

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