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In this study, the biotechnological potential of\n                    <jats:italic>Paenarthrobacter nicotinovorans<\/jats:italic>\n                    ATCC 49919 for selective accumulation of 6-hydroxy-(L)-nicotine (6HLN), the first intermediate of the pyridine pathway, was evaluated under flask and bioreactor conditions. Two metabolic engineering strategies were explored: transcriptional silencing of the 6-hydroxynicotine oxidase gene (\n                    <jats:italic>6hlnO<\/jats:italic>\n                    ) using a CRISPR\/dCas9 system and overexpression of the nicotine dehydrogenase large subunit (\n                    <jats:italic>ndhL<\/jats:italic>\n                    ) to enhance entry flux into the pathway. Among the tested strains, the\n                    <jats:italic>ndhL<\/jats:italic>\n                    -overexpressing derivative exhibited the highest 6HLN accumulation during growth (up to 1.78 g L\u207b\n                    <jats:sup>1<\/jats:sup>\n                    ; space\u2013time yield 104.4 mg L\u207b\n                    <jats:sup>1<\/jats:sup>\n                    h\u207b\n                    <jats:sup>1<\/jats:sup>\n                    ). Resting-cell bioconversion experiments identified 100 g wet cell weight L\u207b\n                    <jats:sup>1<\/jats:sup>\n                    as an optimal biomass loading. Chemical inhibition of 6-hydroxy-L-nicotine oxidase with 0.8 mM ZnSO\n                    <jats:sub>4<\/jats:sub>\n                    further expanded the 6HLN accumulation window without substantially impairing nicotine depletion. Scale-up experiments in a stirred bioreactor demonstrated effective conversion of both pure nicotine and tobacco-derived extracts, with matrix-dependent differences in conversion kinetics. Dissolved oxygen profiles correlated reproducibly with substrate depletion and product turnover, providing a potential online process control signature. Although not achieving near-stoichiometric yields reported for knockout-based systems, the described platform offers a controllable and feedstock-flexible approach for nicotine waste valorization.\n                  <\/jats:p>","DOI":"10.1007\/s42770-026-02064-x","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T11:58:51Z","timestamp":1787054331000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Biotechnological potential of Paenarthrobacter nicotinovorans ATCC 49919 for the microbial conversion of nicotine and tobacco extracts into 6-hydroxynicotine"],"prefix":"10.1007","volume":"57","author":[{"given":"Iustin-Tiberius","family":"Munteanu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Hu\u015fleag","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert Iustin","family":"Susanu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreea-Mihaela","family":"Mle\u015fni\u021b\u0103","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4439-4251","authenticated-orcid":false,"given":"Marius","family":"Mih\u0103\u015fan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,18]]},"reference":[{"key":"2064_CR1","doi-asserted-by":"publisher","unstructured":"Zhang H, Du Y, Guan E, Xu X, Zhang P, Gao L (2025) Meta-omics insights into microbial dynamics and nicotine removal during aerobic fermentation of fresh waste tobacco leaves. 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