Recent advances in the metabolic engineering of microorganisms for the production of 3-hydroxypropionic acid as C3 platform chemical

Chung, Wook-Jin; Liu, Huaiwei; Nisola, Grace M.; Lee, Seung Hwan; Park, Si Jae

doi:10.1007/s00253-013-4802-4

Cited by 66 publications

(28 citation statements)

References 69 publications

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“…5A). Although higher titers have been attained through a two-step conversion from glycerol, our titers are the highest reported for production of 3HP proceeding through E. coli central metabolism (25). These titers were obtained with rich medium in 96-well plates supplemented once with 50 mM glucose at the beginning of fermentation.…”

Section: Resultsmentioning

confidence: 77%

Genetically encoded sensors enable real-time observation of metabolite production

Rogers

Church

2016

Proc. Natl. Acad. Sci. U.S.A.

167

114

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Engineering cells to produce valuable metabolic products is hindered by the slow and laborious methods available for evaluating product concentration. Consequently, many designs go unevaluated, and the dynamics of product formation over time go unobserved. In this work, we develop a framework for observing product formation in real time without the need for sample preparation or laborious analytical methods. We use genetically encoded biosensors derived from small-molecule responsive transcription factors to provide a fluorescent readout that is proportional to the intracellular concentration of a target metabolite. Combining an appropriate biosensor with cells designed to produce a metabolic product allows us to track product formation by observing fluorescence. With individual cells exhibiting fluorescent intensities proportional to the amount of metabolite they produce, high-throughput methods can be used to rank the quality of genetic variants or production conditions. We observe production of several renewable plastic precursors with fluorescent readouts and demonstrate that higher fluorescence is indeed an indicator of higher product titer. Using fluorescence as a guide, we identify process parameters that produce 3-hydroxypropionate at 4.2 g/L, 23-fold higher than previously reported. We also report, to our knowledge, the first engineered route from glucose to acrylate, a plastic precursor with global sales of $14 billion. Finally, we monitor the production of glucarate, a replacement for environmentally damaging detergents, and muconate, a renewable precursor to polyethylene terephthalate and nylon with combined markets of $51 billion, in real time, demonstrating that our method is applicable to a wide range of molecules.biotechnology | directed evolution | biosensor | metabolic engineering | synthetic biology B iological production of valuable products such as pharmaceuticals or renewable chemicals holds the potential to transform the global economy. However, the rate at which bioengineers are able to engineer new living catalysts is hampered by an exceedingly slow design-build-test cycle. We describe a method to accelerate the design-build-test cycle for metabolic engineering by enabling the observation of product formation within microbes as it occurs.Biological production of a desired product is accomplished by guiding a low-cost starting material such as glucose through a series of intracellular enzymatic reactions, ultimately yielding a molecule of economic interest. The choice of culture conditions, the creation of enzyme variants, and the tuning of endogenous cellular metabolism create a vast universe of potential designs. Because of the complexity of biology, appropriate genetic designs and culture conditions are not known a priori. Even sophisticated modeling paradigms can result in very large design spaces (1, 2). Evaluating genetic designs or modulating process parameters to achieve a desired outcome is therefore a major bottleneck in the bioengineering design-build-test cycle.Current methods...

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Section: Resultsmentioning

confidence: 77%

Genetically encoded sensors enable real-time observation of metabolite production

Rogers

Church

2016

Proc. Natl. Acad. Sci. U.S.A.

167

114

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“…Microbial conversion of glycerol into 3-HP has been reported most extensively in bacterial systems, such as E. coli and Klebsiella pneumonia [24][25][26]. Both CoA-dependent and CoA-independent pathways have been described for the production of 3-HP with glycerol as carbon substrate, nevertheless, the CoA-dependent pathway has been less explored as it is more complicated compared with the CoA-independent pathway.…”

Section: Production Of 3-hp From Glycerolmentioning

confidence: 99%

Biobased organic acids production by metabolically engineered microorganisms

Chen

Nielsen

2016

Current Opinion in Biotechnology

143

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“…Therefore, several PHB copolymers have been produced from low-cost glucose via engineered metabolic pathways, including poly(3-hydroxybutyrate-co-lactide) [P(3HB-co-LA)] or PHBLA [16,17], poly(3-hydroxybutyrate-co-3-hydroxypropionate) [P(3HB-co-3HP)] or P3HP3HB [18,19], P3HB4HB [20], poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] or PHBV [21,22], poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] or PHBHHx [23,24], and poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) [P(3HB-co-3HA)] [20] (Table 1). Several synthetic pathways have been proposed and realized for the production of poly-3-hydroxypropionate (P3HP) from glucose or glycerol [18,[25][26][27]. All these PHAs were possible because the PHA synthase PhaC1 of Ralstonia eutropha can actively polymerize scl PHA monomers that differ from its native substrate [28].…”

Section: Engineering Bacterial Pha Synthesis Mechanismsmentioning

confidence: 99%