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Exploiting Hydrogenophaga pseudoflava for aerobic syngas-based production of chemicals.
Metab Eng 2019; 55:220-230ME

Abstract

Gasification is a suitable technology to generate energy-rich synthesis gas (syngas) from biomass or waste streams, which can be utilized in bacterial fermentation processes for the production of chemicals and fuels. Established microbial processes currently rely on acetogenic bacteria which perform an energetically inefficient anaerobic CO oxidation and acetogenesis potentially hampering the biosynthesis of complex and ATP-intensive products. Since aerobic oxidation of CO is energetically more favorable, we exploit in this study the Gram-negative β-proteobacterium Hydrogenophaga pseudoflava DSM1084 as novel host for the production of chemicals from syngas. We sequenced and annotated the genome of H. pseudoflava and established a genetic engineering toolbox, which allows markerless chromosomal modification via the pk19mobsacB system and heterologous gene expression on pBBRMCS2-based plasmids. The toolbox was extended by identifying strong endogenous promotors such as PgapA2 which proved to yield high expression under heterotrophic and autotrophic conditions. H. pseudoflava showed relatively fast heterotrophic growth in complex and minimal medium with sugars and organic acids which allows convenient handling in lab routines. In autotrophic bioreactor cultivations with syngas, H. pseudoflava exhibited a growth rate of 0.06 h-1 and biomass specific uptakes rates of 14.2 ± 0.3 mmol H2 gCDW-1 h-1, 73.9 ± 1.8 mmol CO gCDW-1 h-1, and 31.4 ± 0.3 mmol O2 gCDW-1 h-1. As proof of concept, we engineered the carboxydotrophic bacterium for the aerobic production of the C15 sesquiterpene (E)-α-bisabolene from the C1 carbon source syngas by heterologous expression of the (E)-α-bisabolene synthase gene agBIS. The resulting strain H. pseudoflava (pOCEx1:agBIS) produced 59 ± 8 μg (E)-α-bisabolene L-1 with a volumetric productivity Qp of 1.2 ± 0.2 μg L-1 h-1 and a biomass-specific productivity qp of 13.1 ± 0.6 μg gCDW-1 h-1. The intrinsic properties and the genetic repertoire of H. pseudoflava make this carboxydotrophic bacterium a promising candidate for future aerobic production processes to synthesize more complex or ATP-intensive chemicals from syngas.

Authors+Show Affiliations

Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.Center for Biotechnology, Bielefeld University, Universitätsstraβe 27, 33615, Bielefeld, Germany.Department of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.Institute of Microbiology and Biotechnology, University of Ulm, 89069, Ulm, Germany; Microbial Biotechnology, Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Straubing, Germany.Institute of Microbiology and Biotechnology, University of Ulm, 89069, Ulm, Germany.Institute of Microbiology and Biotechnology, University of Ulm, 89069, Ulm, Germany.Department of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.Center for Biotechnology, Bielefeld University, Universitätsstraβe 27, 33615, Bielefeld, Germany.Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany.Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany; Microbial Biotechnology, Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Straubing, Germany. Electronic address: bastian.blombach@tum.de.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31319152

Citation

Grenz, Sebastian, et al. "Exploiting Hydrogenophaga Pseudoflava for Aerobic Syngas-based Production of Chemicals." Metabolic Engineering, vol. 55, 2019, pp. 220-230.
Grenz S, Baumann PT, Rückert C, et al. Exploiting Hydrogenophaga pseudoflava for aerobic syngas-based production of chemicals. Metab Eng. 2019;55:220-230.
Grenz, S., Baumann, P. T., Rückert, C., Nebel, B. A., Siebert, D., Schwentner, A., ... Blombach, B. (2019). Exploiting Hydrogenophaga pseudoflava for aerobic syngas-based production of chemicals. Metabolic Engineering, 55, pp. 220-230. doi:10.1016/j.ymben.2019.07.006.
Grenz S, et al. Exploiting Hydrogenophaga Pseudoflava for Aerobic Syngas-based Production of Chemicals. Metab Eng. 2019 Jul 15;55:220-230. PubMed PMID: 31319152.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Exploiting Hydrogenophaga pseudoflava for aerobic syngas-based production of chemicals. AU - Grenz,Sebastian, AU - Baumann,Philipp T, AU - Rückert,Christian, AU - Nebel,Bernd A, AU - Siebert,Daniel, AU - Schwentner,Andreas, AU - Eikmanns,Bernhard J, AU - Hauer,Bernhard, AU - Kalinowski,Jörn, AU - Takors,Ralf, AU - Blombach,Bastian, Y1 - 2019/07/15/ PY - 2019/03/21/received PY - 2019/06/25/revised PY - 2019/07/12/accepted PY - 2019/7/19/pubmed PY - 2019/7/19/medline PY - 2019/7/19/entrez KW - Bisabolene KW - Carbon dioxide KW - Carbon monoxide KW - Hydrogen KW - Hydrogenophaga pseudoflava KW - Syngas SP - 220 EP - 230 JF - Metabolic engineering JO - Metab. Eng. VL - 55 N2 - Gasification is a suitable technology to generate energy-rich synthesis gas (syngas) from biomass or waste streams, which can be utilized in bacterial fermentation processes for the production of chemicals and fuels. Established microbial processes currently rely on acetogenic bacteria which perform an energetically inefficient anaerobic CO oxidation and acetogenesis potentially hampering the biosynthesis of complex and ATP-intensive products. Since aerobic oxidation of CO is energetically more favorable, we exploit in this study the Gram-negative β-proteobacterium Hydrogenophaga pseudoflava DSM1084 as novel host for the production of chemicals from syngas. We sequenced and annotated the genome of H. pseudoflava and established a genetic engineering toolbox, which allows markerless chromosomal modification via the pk19mobsacB system and heterologous gene expression on pBBRMCS2-based plasmids. The toolbox was extended by identifying strong endogenous promotors such as PgapA2 which proved to yield high expression under heterotrophic and autotrophic conditions. H. pseudoflava showed relatively fast heterotrophic growth in complex and minimal medium with sugars and organic acids which allows convenient handling in lab routines. In autotrophic bioreactor cultivations with syngas, H. pseudoflava exhibited a growth rate of 0.06 h-1 and biomass specific uptakes rates of 14.2 ± 0.3 mmol H2 gCDW-1 h-1, 73.9 ± 1.8 mmol CO gCDW-1 h-1, and 31.4 ± 0.3 mmol O2 gCDW-1 h-1. As proof of concept, we engineered the carboxydotrophic bacterium for the aerobic production of the C15 sesquiterpene (E)-α-bisabolene from the C1 carbon source syngas by heterologous expression of the (E)-α-bisabolene synthase gene agBIS. The resulting strain H. pseudoflava (pOCEx1:agBIS) produced 59 ± 8 μg (E)-α-bisabolene L-1 with a volumetric productivity Qp of 1.2 ± 0.2 μg L-1 h-1 and a biomass-specific productivity qp of 13.1 ± 0.6 μg gCDW-1 h-1. The intrinsic properties and the genetic repertoire of H. pseudoflava make this carboxydotrophic bacterium a promising candidate for future aerobic production processes to synthesize more complex or ATP-intensive chemicals from syngas. SN - 1096-7184 UR - https://www.unboundmedicine.com/medline/citation/31319152/Exploiting_Hydrogenophaga_pseudoflava_for_aerobic_syngas-based_production_of_chemicals L2 - https://linkinghub.elsevier.com/retrieve/pii/S1096-7176(19)30124-7 DB - PRIME DP - Unbound Medicine ER -