An Efficient Transformation Method for the Bioplastic‐Producing “Knallgas” Bacterium <i>Ralstonia eutropha</i> H16

Tee, Kang Lan; Grinham, James; Othusitse, Arona Martin; Gonzalez‐Villanueva, Miriam; Johnson, Abayomi Oluwanbe; Wong, Tuck Seng

doi:10.1002/biot.201700081

Cited by 27 publications

(27 citation statements)

References 26 publications

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“…To determine their effects on growth in glycerol, plasmids were transformed into C. necator H16 by electroporation [36]. A pre-culture was prepared from a single colony in MSM with 1.00% ( w / v ) gluconate.…”

Section: Methodsmentioning

confidence: 99%

Adaptive Laboratory Evolution of Cupriavidus necator H16 for Carbon Co-Utilization with Glycerol

Gonzalez‐Villanueva

Galaiya

Staniland

et al. 2019

IJMS

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Cupriavidus necator H16 is a non-pathogenic Gram-negative betaproteobacterium that can utilize a broad range of renewable heterotrophic resources to produce chemicals ranging from polyhydroxybutyrate (biopolymer) to alcohols, alkanes, and alkenes. However, C. necator H16 utilizes carbon sources to different efficiency, for example its growth in glycerol is 11.4 times slower than a favorable substrate like gluconate. This work used adaptive laboratory evolution to enhance the glycerol assimilation in C. necator H16 and identified a variant (v6C6) that can co-utilize gluconate and glycerol. The v6C6 variant has a specific growth rate in glycerol 9.5 times faster than the wild-type strain and grows faster in mixed gluconate–glycerol carbon sources compared to gluconate alone. It also accumulated more PHB when cultivated in glycerol medium compared to gluconate medium while the inverse is true for the wild-type strain. Through genome sequencing and expression studies, glycerol kinase was identified as the key enzyme for its improved glycerol utilization. The superior performance of v6C6 in assimilating pure glycerol was extended to crude glycerol (sweetwater) from an industrial fat splitting process. These results highlight the robustness of adaptive laboratory evolution for strain engineering and the versatility and potential of C. necator H16 for industrial waste glycerol valorization.

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Section: Methodsmentioning

confidence: 99%

Adaptive Laboratory Evolution of Cupriavidus necator H16 for Carbon Co-Utilization with Glycerol

Gonzalez‐Villanueva

Galaiya

Staniland

et al. 2019

IJMS

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“…The low electroporation efficiency in C. necator can be circumvented in several ways. Some groups have been able to electroporate plasmids into C. necator with high-molecular-weight buffers, often containing sucrose [65]. However, Xiong and coworkers identified several restriction endonucleases encoded by the C. necator genome and created knockouts of five putative restriction systems in C. necator.…”

Section: Synthetic Biology Tools For C Necatormentioning

confidence: 99%

“…Plasmids with the RP4 (RK2) origin can also be used [68]. Additional plasmids that have been used for stable gene expression in C. necator include pMOL28, RP4, RSF1010, and pSa as well as pBHR1, and modular vectors are freely available with these features through the Standard European Vector Architecture (SEVA) [65,69]. Endogenous inducible promoters can be used in certain contexts, but are coupled to the metabolic state of the cell.…”

Section: Synthetic Biology Tools For C Necatormentioning

confidence: 99%

Metabolic Engineering of Cupriavidus necator H16 for Sustainable Biofuels from CO2

Panich

Fong

Singer

2021

Trends in Biotechnology

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Decelerating global warming is one of the predominant challenges of our time and will require conversion of CO 2 to usable products and commodity chemicals. Of particular interest is the production of fuels, because the transportation sector is a major source of CO 2 emissions. Here, we review recent technological advances in metabolic engineering of the hydrogen-oxidizing bacterium Cupriavidus necator H16, a chemolithotroph that naturally consumes CO 2 to generate biomass. We discuss recent successes in biofuel production using this organism, and the implementation of electrolysis/artificial photosynthesis approaches that enable growth of C. necator using renewable electricity and CO 2 . Last, we discuss prospects of improving the nonoptimal growth of C. necator in ambient concentrations of CO 2 . Mitigation of Atmospheric CO 2 with BioconversionSatisfying current and future global energy demand while allowing favorable environmental outcomes will require innovative solutions in the fuel sector. Although portions of the transportation infrastructure can be electrified (such as automobile transport), energy-dense carbon-based fuels will continue to be required for aviation, long-distance trucking, rocketry, maritime shipping, and industrial operations. Petroleum-based fuels are finite, given that BP and the International Energy Agency (IEA) project that petroleum sources are likely to be depleted between 2050 and 2070 using current oil extraction technologies i-iii . Climate change caused by an increase in CO 2 (and CH 4 ) levels in Earth's atmosphere has brought cataclysmic weather events and has decimated ocean habitats in recent years, a trend that will likely continue through our lifetimes given that CO 2 is being emitted into the atmosphere at a rate of ≈32 billion tons of CO 2 per year [1-6] iv . Therefore, sustainable and economically viable bioconversion of CO 2 to fuels and commodity chemicals should be realized within the next decade. A small but significant mitigation of anthropogenic CO 2 release can be achieved by using CO 2 from atmospheric air as a feedstock to produce biofuels in microorganisms: if 10% of global aviation fuel (accounting for~2.6% of total CO 2 emissions [7]) were replaced with Sustainable Aviation Fuels (SAFs) from Cupriavidus necator at a titer of 1 g/l and assuming 90 g/l biomass accumulation [8],~500 000 tons of sustainable fuel per year could be produced from recycled CO 2 (at a carbon density of 0.8 kg/l). In addition, the accumulated biomass would sequester~80 million tons of CO 2 per year (mitigating ≈0.25% of emissions).CO 2 is an ideal feedstock for the production of biofuels because it is an inexpensive, nontoxic, and abundant starting material (≈850 billion tons of CO 2 are currently present in the atmosphere). Furthermore, CO 2 is noncompetitive with the global food supply chain. Technoeconomic analysis indicates that CO 2 -derived biofuel production could be a US$10-250 billion industry by 2030 v . However, CO 2 is dilute in the atmosphere (≈0.04% CO 2 by volume) and ...

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“…SCCGE1002. These bacterial species are found in soil and have the ability to nodulate in legumes and to accumulate polyhydroxybutyrate (PHB) under stress conditions (Ormeño‐Orrillo et al., ; Tee et al., ). On the other hand, the regions comprising T6SS‐2 and T6SS‐3 show a higher identity to B. cenocepacia and P .…”

Section: Resultsmentioning

confidence: 99%

Comparative genomics of Paraburkholderia kururiensis and its potential in bioremediation, biofertilization, and biocontrol of plant pathogens

et al. 2019

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Burkholderia harbors versatile Gram‐negative species and is β‐Proteobacteria. Recently, it was proposed to split the genus in two main branches: one of animal and plant pathogens and another, Paraburkholderia, harboring environmental and plant‐beneficial species. Currently, Paraburkholderia comprises more than 70 species with ability to occupy very diverse environmental niches. Herein, we sequenced and analyzed the genome of Paraburkholderia kururiensis type strain KP23T, and compared to P. kururiensis M130, isolated in Brazil, and P. kururiensis susbp. thiooxydans, from Korea. This study focused on the gene content of the three genomes with special emphasis on their potential of plant‐association, biocontrol, and bioremediation. The comparative analyses revealed several genes related to plant benefits, including biosynthesis of IAA, ACC deaminase, multiple efflux pumps, dioxygenases, and degradation of aromatic compounds. Importantly, a range of genes for protein secretion systems (type III, IV, V, and VI) were characterized, potentially involved in P. kururiensis well documented ability to establish endophytic association with plants. These findings shed light onto bacteria‐plant interaction mechanisms at molecular level, adding novel information that supports their potential application in bioremediation, biofertilization, and biocontrol of plant pathogens. P. kururiensis emerges as a promising model to investigate adaptation mechanisms in different ecological niches.

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An Efficient Transformation Method for the Bioplastic‐Producing “Knallgas” Bacterium Ralstonia eutropha H16

Cited by 27 publications

References 26 publications

Adaptive Laboratory Evolution of Cupriavidus necator H16 for Carbon Co-Utilization with Glycerol

Adaptive Laboratory Evolution of Cupriavidus necator H16 for Carbon Co-Utilization with Glycerol

Metabolic Engineering of Cupriavidus necator H16 for Sustainable Biofuels from CO2

Comparative genomics of Paraburkholderia kururiensis and its potential in bioremediation, biofertilization, and biocontrol of plant pathogens

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