Metabolic regulation of NADH supply and hydrogen production in Enterobacter aerogenes by multi-gene engineering

Bai, Ruoxuan; Chu, Wanying; Qiao, Zimu; Lü, Ping; Jiang, Ke; Xu, Yizhuang; Yang, Jiayao; Gao, Ting; Xu, Fangxu; Zhao, Hongxin

doi:10.1016/j.ijhydene.2022.10.015

Cited by 8 publications

(2 citation statements)

References 43 publications

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“…putida strain AA7 ( Ppu ) where genetic tools/methods developed in E. coli have been shown to be adaptable for Enterobacter strains − and a plethora of tools developed in other P. putida strains that may be adapted to Ppu . − The rest of the SynCom, Stenotrophomonas maltophilia strain AA1 ( Sma ), Brucella pituitosa strain AA2 ( Bpi , formerly Ochrobactrum pituitosum), Curtobacterium pusillum strain AA3 ( CPU ), Chryseobacterium indologenes strain AA5 ( Cin ), and Herbaspirillum robiniae strain AA6 ( Hro , formerly Herbaspirillum frisingense), vary in the availability of genetic tools.…”

Section: Introductionmentioning

confidence: 99%

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for Studying Plant–Microbe Interactions

van Schaik,

Li,

Cheadle

et al. 2023

ACS Synth. Biol.

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Sustainably enhancing crop production is a global necessity to meet the escalating demand for staple crops while sustainably managing their associated carbon/nitrogen inputs. Leveraging plant-associated microbiomes is a promising avenue for addressing this demand. However, studying these communities and engineering them for sustainable enhancement of crop production have remained a challenge due to limited genetic tools and methods. In this work, we detail the development of the Maize Root Microbiome ToolKit (MRMTK), a rapid Modular Cloning (MoClo) toolkit that only takes 2.5 h to generate desired constructs (5400 potential plasmids) that replicate and express heterologous genes in Enterobacter ludwigii strain AA4 (Elu), Pseudomonas putida strain AA7 (Ppu), Herbaspirillum robiniae strain AA6 (Hro), Stenotrophomonas maltophilia strain AA1 (Sma), and Brucella pituitosa strain AA2 (Bpi), which comprise a model maize root synthetic community (SynCom). In addition to these genetic tools, we describe a highly efficient transformation protocol (107–109 transformants/μg of DNA) 1 for each of these strains. Utilizing this highly efficient transformation protocol, we identified endogenous Expression Sequences (ES; promoter and ribosomal binding sites) for each strain via genomic promoter trapping. Overall, MRMTK is a scalable and adaptable platform that expands the genetic engineering toolbox while providing a standardized, high-efficiency transformation method across a diverse group of root commensals. These results unlock the ability to elucidate and engineer plant–microbe interactions promoting plant growth for each of the 5 bacterial strains in this study.

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Section: Introductionmentioning

confidence: 99%

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for Studying Plant–Microbe Interactions

van Schaik,

Li,

Cheadle

et al. 2023

ACS Synth. Biol.

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show abstract

“…Genetic tools are available for two SynCom species: Enterobacter ludwigii strain AA4 (Elu, formerly E. cloacae) and Pseudomonas putida strain AA7 (Ppu) where genetic tools/methods developed in E. coli have been shown to be adaptable for Enterobacter strains (44)(45)(46) and a plethora of tools developed in other Pseudomonas putida strains that may be adapted to strain AA7 (16)(17)(18)(19). The rest of the SynCom, Stenotrophomonas maltophilia strain AA1 (Sma), Brucella pituitosa strain AA2 (Bpi, formerly Ochrobactrum pituitosum), Curtobacterium pusillum strain AA3 (Cpu), Chryseobacterium indologenes strain AA5 (Cin) and Herbaspirillum robiniae strain AA6 (Hro, formerly H. frisingense), vary in the availability of genetic tools.…”

Section: Introductionmentioning

confidence: 99%

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for studying Plant-Microbe Interactions

Schaik

Cheadle

et al. 2023

Preprint

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Sustainably enhancing crop production is a necessity given the increasing demands for staple crops and their associated carbon/nitrogen inputs. Plant-associated microbiomes offer one avenue for addressing this demand; however, studying these communities and engineering them has remained a challenge due to limited genetic tools and methods. In this work, we detail the development of the Maize Root ToolKit (MRTK); a rapid Modular Cloning (MoClo) toolkit that only takes 2.5 hours to generate desired constructs (5400 potential plasmids) that replicate and express heterologous genes in Enterobacter ludwigii strain AA4 (Elu), Pseudomonas putida AA7 (Ppu), Herbaspirillum robiniae strain AA6 (Hro), Stenotrophomonas maltophilia strain AA1 (Sma) and Brucella pituitosa strain AA2 (Bpi) which comprise a model maize root synthetic community (SynCom). In addition to these genetic tools, we describe a highly efficient transformation protocol (10^7-10^9 transformants/ug of DNA) for each of these strains. Utilizing this highly efficient transformation protocol, we identified endogenous expression sequences for each strain (ES; promoter and ribosomal binding sites) via genomic promoter trapping. Overall, the MRTK is a scalable platform that expands the genetic engineering toolbox while providing a standardized, high efficiency transformation method that can be implemented across a diverse group of root commensals. These results unlock the ability to elucidate and engineer plant-microbe interactions promoting plant growth for each of the 5 bacterial strains in this study.

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Boosting solar hydrogen production via electrostatic interaction mediated E. coli-TiO2−x biohybrid system

Lv,

Huang,

Gao

et al. 2024

Nano Res.

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Metabolic regulation of NADH supply and hydrogen production in Enterobacter aerogenes by multi-gene engineering

Cited by 8 publications

References 43 publications

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for Studying Plant–Microbe Interactions

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for Studying Plant–Microbe Interactions

Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and Identification of Potential Bacterial Chassis for studying Plant-Microbe Interactions

Boosting solar hydrogen production via electrostatic interaction mediated E. coli-TiO2−x biohybrid system

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