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 H2) or from C1-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 OM5 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 H2 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
clusters encoding proteins required for autotrophic growth (CO2 fixation via Calvin–Benson–Bassham
cycle), for CO metabolism (CO dehydrogenase), and for H2 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.