{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:12:45Z","timestamp":1785888765650,"version":"3.56.0"},"reference-count":115,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T00:00:00Z","timestamp":1768867200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T00:00:00Z","timestamp":1768867200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000015","name":"U.S. Department of Energy","doi-asserted-by":"publisher","award":["DE-SC0018409"],"award-info":[{"award-number":["DE-SC0018409"]}],"id":[{"id":"10.13039\/100000015","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["T32GM135066"],"award-info":[{"award-number":["T32GM135066"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008982","name":"National Science Foundation","doi-asserted-by":"publisher","award":["2137424"],"award-info":[{"award-number":["2137424"]}],"id":[{"id":"10.13039\/501100008982","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mSystems"],"published-print":{"date-parts":[[2026,1,20]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Genetically engineered microbes have the potential to increase efficiency in the bioeconomy by overcoming growth-limiting production stress. Screens of gene perturbation libraries against production stressors can identify high-value engineering targets, but follow-up experiments needed to guard against false positives are slow and resource-intensive. In principle, the use of orthogonal gene perturbation approaches could increase recovery of true positives over false positives because the strengths of one technique compensate for the weaknesses of the other, but, in practice, two parallel screens are rarely performed at the genome scale. Here, we screen genome-scale CRISPRi (CRISPR interference) knockdown and transposon insertion libraries of the bioenergy-relevant Alphaproteobacterium,\n                      <jats:italic toggle=\"yes\">Zymomonas mobilis<\/jats:italic>\n                      , against growth inhibitors commonly found in deconstructed plant material. Integrating data from the two gene perturbation techniques, we established an approach for defining engineering targets with high specificity. This allowed us to identify all known genes in the cytochrome\n                      <jats:italic toggle=\"yes\">bc<\/jats:italic>\n                      <jats:sub>1<\/jats:sub>\n                      and cytochrome\n                      <jats:italic toggle=\"yes\">c<\/jats:italic>\n                      synthesis pathway as potential targets for engineering resistance to phenolic acids under anaerobic conditions, a subset of which we validated using precise gene deletions. Strikingly, this finding is specific to the cytochrome\n                      <jats:italic toggle=\"yes\">bc<\/jats:italic>\n                      <jats:sub>1<\/jats:sub>\n                      and cytochrome\n                      <jats:italic toggle=\"yes\">c<\/jats:italic>\n                      pathway and does not extend to other branches of the electron transport chain. We further show that exposure of\n                      <jats:italic toggle=\"yes\">Z. mobilis<\/jats:italic>\n                      to ferulic acid causes substantial remodeling of the cell envelope proteome, as well as the downregulation of TonB-dependent transporters. Our work provides a generalizable strategy for identifying high-value engineering targets from gene perturbation screens that is broadly applicable.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Engineering microorganisms to tolerate harsh production conditions will contribute to increased bioproduct yields. In this study, we systematically identified\n                        <jats:italic toggle=\"yes\">Zymomonas mobilis<\/jats:italic>\n                        genes that confer resistance or susceptibility to chemical stressors found in deconstructed plant material. We used complementary genetic techniques to cross-validate these genes at scale, providing a widely applicable method for precisely identifying genetic alterations that increase chemical resilience. We discovered genetic modifications that improve anaerobic growth of\n                        <jats:italic toggle=\"yes\">Z. mobilis<\/jats:italic>\n                        in the presence of inhibitory chemicals found in renewable plant-based feedstocks. These results have implications for engineering robust production strains to support efficient and resilient bioproduction. Our methodologies can be broadly applied to understand microbial responses to chemicals across systems, paving the way for developments in biomanufacturing, therapeutics, and agriculture.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/msystems.01001-25","type":"journal-article","created":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T14:00:16Z","timestamp":1764856816000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":4,"title":["Orthogonal chemical genomics approaches reveal genomic targets for increasing anaerobic chemical tolerance in\n                    <i>Zymomonas mobilis<\/i>"],"prefix":"10.1128","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-0778-7144","authenticated-orcid":true,"given":"Jacob 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