Systems-Level Analysis of Oxygen Exposure in Zymomonas mobilis : Implications for Isoprenoid Production

Journal Article · 12 February 2019 · mSystems

DOI: https://doi.org/10.1128/msystems.00284-18 · OSTI ID:1494210

Martien, Julia I. ^[1]; Hebert, Alexander S. ^[1]; Stevenson, David M. ^[1]; Regner, Matthew R. ^[1]; Khana, Daven B. ^[1]; Coon, Joshua J. ^[2]; Amador-Noguez, Daniel ^[3]; Vickers, Claudia ^[3]

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1. Univ. of Wisconsin, Madison, WI (United States)
2. Univ. of Wisconsin, Madison, WI (United States); Morgridge Inst. for Research, Madison, WI (United States)
3. Morgridge Institute for Research, Madison, Wisconsin, USA

Zymomonas mobilisis an aerotolerant anaerobe and prolific ethanologen with attractive characteristics for industrial bioproduct generation. However, there is currently insufficient knowledge of the impact that environmental factors have on flux through industrially relevant biosynthetic pathways. Here, we examined the effect of oxygen exposure on metabolism and gene expression inZ. mobilisby combining targeted metabolomics, mRNA sequencing, and shotgun proteomics. We found that exposure to oxygen profoundly influenced metabolism, inducing both transient metabolic bottlenecks and long-term metabolic remodeling. In particular, oxygen induced a severe but temporary metabolic bottleneck in the methyl erythritol 4-phosphate pathway for isoprenoid biosynthesis caused by oxidative damage to the iron-sulfur cofactors of the final two enzymes in the pathway. This bottleneck was resolved with minimal changes in expression of isoprenoid biosynthetic enzymes. Instead, it was associated with pronounced upregulation of enzymes related to iron-sulfur cluster maintenance and biogenesis (i.e., flavodoxin reductase and thesufoperon). We also detected major changes in glucose utilization in the presence of oxygen. Specifically, we observed increased gluconate production following exposure to oxygen, accounting for 18% of glucose uptake. Our results suggest that under aerobic conditions, electrons derived from the oxidation of glucose to gluconate are diverted to the electron transport chain, where they can minimize oxidative damage by reducing reactive oxygen species such as H₂O₂. This model is supported by the simultaneous upregulation of three membrane-bound dehydrogenases, cytochromecperoxidase, and a cytochromebdoxidase following exposure to oxygen. Microbially generated biofuels and bioproducts have the potential to provide a more environmentally sustainable alternative to fossil-fuel-derived products. In particular, isoprenoids, a diverse class of natural products, are chemically suitable for use as high-grade transport fuels and other commodity molecules. However, metabolic engineering for increased production of isoprenoids and other bioproducts is limited by an incomplete understanding of factors that control flux through biosynthetic pathways. Here, we examined the native regulation of the isoprenoid biosynthetic pathway in the biofuel producerZymomonas mobilis. We leveraged oxygen exposure as a means to perturb carbon flux, allowing us to observe the formation and resolution of a metabolic bottleneck in the pathway. Our multi-omics analysis of this perturbation enabled us to identify key auxiliary enzymes whose expression correlates with increased production of isoprenoid precursors, which we propose as potential targets for future metabolic engineering.

• Research Organization: Univ. of Wisconsin, Madison, WI (United States)
• Sponsoring Organization: USDOE Office of Science (SC)
• Grant/Contract Number: FC02-07ER64494; SC0018409
• OSTI ID: 1494210
• Alternate ID(s): OSTI ID: 1609268
• Journal Information: mSystems, Vol. 4, Issue 1; ISSN 2379-5077
• Publisher: American Society for MicrobiologyCopyright Statement
• Country of Publication: United States
• Language: English