Genetic Engineering of Oligotropha carboxidovorans Strain OM5-A Promising Candidate for the Aerobic Utilization of Synthesis Gas

Siebert D, Busche T, Metz AY, Smaili M, Queck BAW, Kalinowski J, Eikmanns BJ (2020)
ACS SYNTHETIC BIOLOGY 9(6): 1426-1440.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Autor*in
Siebert, Daniel; Busche, TobiasUniBi; Metz, Aline Y.; Smaili, Medina; Queck, Bastian A. W.; Kalinowski, JörnUniBi; Eikmanns, Bernhard J.
Abstract / Bemerkung
Due to climate change and worldwide pollution, development of highly sustainable routes for industrial production of basic and specialty chemicals is critical nowadays. One possible approach is the use of CO2- and CO-utilizing microorganisms in biotechnological processes to produce value-added compounds from synthesis gas (mixtures of CO2, CO, and H-2) or from Cl-containing industrial waste gases. Such syngas fermentation processes have already been established, e.g., biofuel production using strictly anaerobic acetogenic bacteria. However, aerobic processes may be favorable for the formation of more costly (ATP-intensive) products. Oligotropha carboxidovorans strain OMS is an aerobic carboxidotrophic bacterium and potentially a promising candidate for such processes. We here performed RNA-Seq analysis comparing cells of this organism grown heterotrophically with acetate or autotrophically with CO2, CO, and H-2 as carbon and energy source and found a variety of chromosomally and of native plasmid-encoded genes to be highly differentially expressed. In particular, genes and gene dusters encoding proteins required for autotrophic growth (CO2 fixation via Calvin-Benson-Bassham cyde), for CO metabolism (CO dehydrogenase), and for H-2 utilization (hydrogenase), all located on megaplasmid pHCG3, were much higher expressed during autotrophic growth with synthesis gas. Furthermore, we successfully established reproducible transformation of O. carboxidovorans via electroporation and developed gene deletion and gene exchange protocols via two-step recombination, enabling inducible and stable expression of heterologous genes as well as construction of defined mutants of this organism. Thus, this study marks an important step toward metabolic engineering of O. carboxidovorans and effective utilization of C1-containing gases with this organism.
Stichworte
Oligotropha carboxidovorans; RNA-Seq; autotrophic metabolism; genome; editing; carbon monoxide; carbon dioxide
Erscheinungsjahr
2020
Zeitschriftentitel
ACS SYNTHETIC BIOLOGY
Band
9
Ausgabe
6
Seite(n)
1426-1440
ISSN
2161-5063
Page URI
https://pub.uni-bielefeld.de/record/2944774

Zitieren

Siebert D, Busche T, Metz AY, et al. Genetic Engineering of Oligotropha carboxidovorans Strain OM5-A Promising Candidate for the Aerobic Utilization of Synthesis Gas. ACS SYNTHETIC BIOLOGY. 2020;9(6):1426-1440.
Siebert, D., Busche, T., Metz, A. Y., Smaili, M., Queck, B. A. W., Kalinowski, J., & Eikmanns, B. J. (2020). Genetic Engineering of Oligotropha carboxidovorans Strain OM5-A Promising Candidate for the Aerobic Utilization of Synthesis Gas. ACS SYNTHETIC BIOLOGY, 9(6), 1426-1440. doi:10.1021/acssynbio.0c00098
Siebert, D., Busche, T., Metz, A. Y., Smaili, M., Queck, B. A. W., Kalinowski, J., and Eikmanns, B. J. (2020). Genetic Engineering of Oligotropha carboxidovorans Strain OM5-A Promising Candidate for the Aerobic Utilization of Synthesis Gas. ACS SYNTHETIC BIOLOGY 9, 1426-1440.
Siebert, D., et al., 2020. Genetic Engineering of Oligotropha carboxidovorans Strain OM5-A Promising Candidate for the Aerobic Utilization of Synthesis Gas. ACS SYNTHETIC BIOLOGY, 9(6), p 1426-1440.
D. Siebert, et al., “Genetic Engineering of Oligotropha carboxidovorans Strain OM5-A Promising Candidate for the Aerobic Utilization of Synthesis Gas”, ACS SYNTHETIC BIOLOGY, vol. 9, 2020, pp. 1426-1440.
Siebert, D., Busche, T., Metz, A.Y., Smaili, M., Queck, B.A.W., Kalinowski, J., Eikmanns, B.J.: Genetic Engineering of Oligotropha carboxidovorans Strain OM5-A Promising Candidate for the Aerobic Utilization of Synthesis Gas. ACS SYNTHETIC BIOLOGY. 9, 1426-1440 (2020).
Siebert, Daniel, Busche, Tobias, Metz, Aline Y., Smaili, Medina, Queck, Bastian A. W., Kalinowski, Jörn, and Eikmanns, Bernhard J. “Genetic Engineering of Oligotropha carboxidovorans Strain OM5-A Promising Candidate for the Aerobic Utilization of Synthesis Gas”. ACS SYNTHETIC BIOLOGY 9.6 (2020): 1426-1440.

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