RB-TnSeq identifies genetic targets for improved tolerance of Pseudomonas putida towards compounds relevant to lignin conversion

Metabolic Engineering, Journal Year: 2023, Volume and Issue: 77, P. 208 - 218

Published: April 13, 2023

Language: Английский

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Elastomeric vitrimers from designer polyhydroxyalkanoates with recyclability and biodegradability

Science Advances, Journal Year: 2023, Volume and Issue: 9(47)

Published: Nov. 22, 2023

Cross-linked elastomers are stretchable materials that typically not recyclable or biodegradable. Medium-chain-length polyhydroxyalkanoates (mcl-PHAs) soft and ductile, making these bio-based polymers good candidates for biodegradable elastomers. Elasticity is commonly imparted by a cross-linked network structure, covalent adaptable networks have emerged as solution to prepare thermosets via triggered rearrangement of dynamic bonds. Here, we develop chemically installing the within biologically produced mcl-PHAs. Specifically, an engineered strain Pseudomonas putida was used produce mcl-PHAs containing pendent terminal alkenes chemical handles postfunctionalization. Thiol-ene chemistry incorporate boronic ester (BE) cross-links, resulting in PHA-based vitrimers. with BE at low density (<6 mole %) affords soft, elastomeric material demonstrates thermal reprocessability, biodegradability, denetworking end life. The mechanical properties show potential applications including adhesives robotics electronics.

Language: Английский

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Engineering Pseudomonas putida for efficient aromatic conversion to bioproduct using high throughput screening in a bioreactor

Metabolic Engineering, Journal Year: 2021, Volume and Issue: 66, P. 229 - 238

Published: May 6, 2021

Pseudomonas putida KT2440 is an emerging biomanufacturing host amenable for use with renewable carbon streams including aromatics such as para-coumarate. We used a pooled transposon library disrupting nearly all (4,778) non-essential genes to characterize this microbe under common stirred-tank bioreactor parameters quantitative fitness assays. Assessing differential values by monitoring changes in mutant strain abundance identified 33 gene mutants improved across multiple formats. Twenty-one deletion strains from subset were reconstructed, GacA, regulator, TtgB, ABC transporter, and PP_0063, lipid A acyltransferase. Thirteen roles varying cellular functions evaluated conversion of para-coumarate, heterologous bioproduct, indigoidine. Several mutants, the ΔgacA 35 fold showed 8-fold improvement indigoidine production (4.5 g/L, 0.29 g/g, 23% maximum theoretical yield) para-coumarate source.

Language: Английский

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Production of β-ketoadipic acid from glucose in Pseudomonas putida KT2440 for use in performance-advantaged nylons

Cell Reports Physical Science, Journal Year: 2022, Volume and Issue: 3(4), P. 100840 - 100840

Published: April 1, 2022

Biomass-derived chemicals can offer unique chemical functionality relative to petroleum-derived building blocks. To this end, here we report that β-ketoadipic acid (βKA), a C6 diacid with β-ketone group, be used as performance-advantaged replacement for adipic in nylon-6,6 analog. Building on our previous efforts produce shikimate-derived products from carbohydrates, Pseudomonas putida KT2440 is engineered βKA glucose, achieving 26 g/L titer. Following purification, imparts an increase of 69°C above the glass transition temperature and 20% reduced water permeability, equivalent nylon-6,10. Molecular simulations predict enhanced thermal properties result rigidity introduced by β-ketone. Process analysis predicts produced US$1.94/kg sugars, requiring 63% less energy emitting 43% greenhouse gases than fossil-based acid. Overall, study illustrates potential serve useful block bio-based polymers.

Language: Английский

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Dynamic flux regulation for high-titer anthranilate production by plasmid-free, conditionally-auxotrophic strains of Pseudomonas putida

Metabolic Engineering, Journal Year: 2022, Volume and Issue: 73, P. 11 - 25

Published: May 31, 2022

Language: Английский

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