Regulatory response to a hybrid ancestral nitrogenase in Azotobacter vinelandii

Rivier, Alex J.; Myers, Kevin S.; Garcia, Amanda K.; Sobol, Morgan S.; Kaçar, Betül

doi:10.1128/spectrum.02815-23

Title: Regulatory response to a hybrid ancestral nitrogenase in Azotobacter vinelandii

Journal Article · 13 September 2023 · Microbiology Spectrum

DOI: https://doi.org/10.1128/spectrum.02815-23 · OSTI ID:2202765

Rivier, Alex J. ^[1]; Myers, Kevin S. ^[1];

^[1];

^[1]

University of Wisconsin-Madison, WI (United States)

Biological nitrogen fixation, the microbial reduction of atmospheric nitrogen to bioavailable ammonia, represents both a major limitation on biological productivity and a highly desirable engineering target for synthetic biology. However, the engineering of nitrogen fixation requires an integrated understanding of how the gene regulatory dynamics of host diazotrophs respond across sequence-function space of its central catalytic metalloenzyme, nitrogenase. Here, we interrogate this relationship by analyzing the transcriptome of Azotobacter vinelandii engineered with a phylogenetically inferred ancestral nitrogenase protein variant. The engineered strain exhibits reduced cellular nitrogenase activity but recovers wild-type growth rates following an extended lag period. We find that expression of genes within the immediate nitrogen fixation network is resilient to the introduced nitrogenase sequence-level perturbations. Rather the sustained physiological compatibility with the ancestral nitrogenase variant is accompanied by reduced expression of genes that support trace metal and electron resource allocation to nitrogenase. Our results spotlight gene expression changes in cellular processes adjacent to nitrogen fixation as productive engineering considerations to improve compatibility between remodeled nitrogenase proteins and engineered host diazotrophs.

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Research Organization:: Great Lakes Bioenergy Research Center (GLBRC), Madison, WI (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018409

OSTI ID:: 2202765

Journal Information:: Microbiology Spectrum, Vol. 11, Issue 5; ISSN 2165-0497

Publisher:: American Society for MicrobiologyCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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Title: Regulatory response to a hybrid ancestral nitrogenase in Azotobacter vinelandii

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