Abstract
The advantage of using acetogens such as Acetobacterium woodii as biocatalysts converting the cheap substrate and greenhouse gas carbon dioxide (CO2) into value-added chemicals comes together with the disadvantage of a low overall ATP gain due to the bioenergetics associated with the Wood-Ljungdahl pathway. Expanding the product spectrum of recombinant A. woodii strains to compounds with high ATP-demanding biosynthesis is therefore challenging. As a least invasive strategy for improved ATP generation, the exploitation of the arginine deiminase pathway (ADI) was examined under native conditions and via using heterologously expressed genes in A. woodii. Several promoters were analyzed for application of different gene expression levels in A. woodii using β-glucuronidase assays. Heterologous expression of the ADI pathway genes from Clostridium autoethanogenum was controlled using either the constitutive pta-ack promoter from Clostridium ljungdahlii or a tightly regulated tetracycline-inducible promoter Ptet. Unlike constitutive expression, only induced expression of the ADI pathway genes led to a 36% higher maximal OD600 when using arginine (OD600 3.4) as nitrogen source and a 52% lower acetate yield per biomass compared to cells growing with yeast extract as nitrogen source (OD600 2.5). In direct comparison, a 69% higher maximal OD600 and about 60% lower acetate yield per biomass in induced to non-induced recombinant A. woodii cells was noticed when using arginine. Our data suggests the application of the ADI pathway in A. woodii for expanding the product spectrum to compounds with high ATP-demanding biosynthesis.
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Acknowledgments
We thank Eric Ransom and David Weiss for kindly providing plasmid pDSW1728. We thank Sophie Moormann, Julia Nothelfer, Judith Cammerer, and Lucille Exler for conducting enzymatic assays. The provision of the theophylline riboswitch sequence by Gerd Seibold and Christian Rückert is gratefully acknowledged. We thank the reviewers of this paper for constructive criticism and numerous helpful comments.
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This study was funded by the ERA-IB5 program (project CO2CHEM, grant 031A566A).
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Matthias H. Beck declares that he has no conflict of interest. Maximilian Flaiz declares that he has no conflict of interest. Frank R. Bengelsdorf declares that he has no conflict of interest. Peter Dürre declares that he has no conflict of interest.
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Beck, M.H., Flaiz, M., Bengelsdorf, F.R. et al. Induced heterologous expression of the arginine deiminase pathway promotes growth advantages in the strict anaerobe Acetobacterium woodii. Appl Microbiol Biotechnol 104, 687–699 (2020). https://doi.org/10.1007/s00253-019-10248-9
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DOI: https://doi.org/10.1007/s00253-019-10248-9