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Characterization using a polyphasic taxonomic approach, whole-genome sequencing and phylogenomic analyses revealed a close genetic relationship among two strains from Mars 2020 cleanroom floors (179-C4-2-HS, 179-J1A1-HS), one strain from the Agave plant (AT2.8), and another strain from wheat-associated soil (V4I25). All four strains exhibited high 16S rRNA gene sequence similarity (&gt;99.2%) and low average nucleotide identity (ANI) with\n              <jats:italic>Neobacillus niacini<\/jats:italic>\n              NBRC 15566\n              <jats:sup>T<\/jats:sup>\n              , delineating new phylogenetic branches within the genus. Detailed molecular analyses, including\n              <jats:italic>gyr<\/jats:italic>\n              B (90.2%), ANI (86.4%), average amino acid identity (87.8%) phylogenies, digital DNA\u2013DNA hybridization (32.6%), and percentage of conserved proteins (77.7%) indicated significant divergence from\n              <jats:italic>N. niacini<\/jats:italic>\n              NBRC 15566\n              <jats:sup>T<\/jats:sup>\n              . Consequently, these strains have been designated\n              <jats:italic>Neobacillus driksii<\/jats:italic>\n              sp. nov., with the type strain 179-C4-2-HS\n              <jats:sup>T<\/jats:sup>\n              (DSM 115941\n              <jats:sup>T<\/jats:sup>\n              = NRRL B-65665\n              <jats:sup>T<\/jats:sup>\n              ).\n              <jats:italic>N. driksii<\/jats:italic>\n              grew at 4\u00b0C to 45\u00b0C, pH range of 6.0 to 9.5, and 0.5% to 5% NaCl. The major cellular fatty acids are iso-C\n              <jats:sub>15:0<\/jats:sub>\n              and anteiso-C\n              <jats:sub>15:0<\/jats:sub>\n              . The dominant polar lipids include diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and an unidentified aminolipid. Metagenomic analysis within NASA cleanrooms revealed that\n              <jats:italic>N. driksii<\/jats:italic>\n              is scarce (17 out of 236 samples). Genes encoding the biosynthesis pathway for lasso peptides were identified in all\n              <jats:italic>N. driksii<\/jats:italic>\n              strains and are not commonly found in other\n              <jats:italic>Neobacillus<\/jats:italic>\n              species, except in 7 out of 26 recognized species. This study highlights the unique metabolic capabilities of\n              <jats:italic>N. driksii<\/jats:italic>\n              , underscoring their potential in antimicrobial research and biotechnology.\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              The microbial surveillance of the Mars 2020 assembly cleanroom led to the isolation of novel\n              <jats:italic>N. driksii<\/jats:italic>\n              with potential applications in cleanroom environments, such as hospitals, pharmaceuticals, semiconductors, and aeronautical industries.\n              <jats:italic>N. driksii<\/jats:italic>\n              genomes were found to possess genes responsible for producing lasso peptides, which are crucial for antimicrobial defense, communication, and enzyme inhibition. Isolation of\n              <jats:italic>N. driksii<\/jats:italic>\n              from cleanrooms, Agave plants, and dryland wheat soils, suggested niche-specific ecology and resilience under various environmentally challenging conditions. The discovery of potent antimicrobial agents from novel\n              <jats:italic>N. driksii<\/jats:italic>\n              underscores the importance of genome mining and the isolation of rare microorganisms. Bioactive gene clusters potentially producing nicotianamine-like siderophores were found in\n              <jats:italic>N. driksii<\/jats:italic>\n              genomes. These siderophores can be used for bioremediation to remove heavy metals from contaminated environments, promote plant growth by aiding iron uptake in agriculture, and treat iron overload conditions in medical applications.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.01376-24","type":"journal-article","created":{"date-parts":[[2024,11,29]],"date-time":"2024-11-29T15:47:07Z","timestamp":1732895227000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":10,"title":["<i>Neobacillus driksii<\/i>\n            sp. nov. isolated from a Mars 2020 spacecraft assembly facility and genomic potential for lasso peptide production in\n            <i>Neobacillus<\/i>"],"prefix":"10.1128","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4445-7080","authenticated-orcid":true,"given":"Asif","family":"Hameed","sequence":"first","affiliation":[{"name":"Division of Microbiology and Biotechnology, 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