AUTHOR=Mhatre Snehit , Singh Nitin K. , Wood Jason M. , Parker Ceth W. , Pukall Rüdiger , Verbarg Susanne , Tindall Brian J. , Neumann-Schaal Meina , Venkateswaran Kasthuri 

TITLE=Description of Chloramphenicol Resistant Kineococcus rubinsiae sp. nov. Isolated From a Spacecraft Assembly Facility

JOURNAL=Frontiers in Microbiology

VOLUME=11

YEAR=2020

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.01957

DOI=10.3389/fmicb.2020.01957

ISSN=1664-302X

ABSTRACT=<p>A Gram-positive, coccoid, motile, aerobic bacterium, designated strain B12<sup>T</sup> was isolated from a Jet Propulsion Laboratory spacecraft assembly cleanroom, Pasadena, CA, United States. Strain B12<sup>T</sup> was resistant to chloramphenicol (100 μg/mL), and is a relatively slow grower (3–5 days optimal). Strain B12<sup>T</sup> was found to grow optimally at 28 to 32°C, pH 7 to 8, and 0.5% NaCl. Fatty acid methyl ester analysis showed that the major fatty acid of the strain B12<sup>T</sup> was anteiso C<sub>15</sub><sub>:</sub><sub>0</sub> (66.3%), which is also produced by other <italic>Kineococcus</italic> species. However, arachidonic acid (C<sub>20</sub><sub>:</sub><sub>4</sub> ω6,9,12,16c) was present in strain B12<sup>T</sup> and <italic>Kineococcus glutinatus</italic> YIM 75677<sup>T</sup> but absent in all other <italic>Kineococcus</italic> species. 16S rRNA analysis revealed that strain B12<sup>T</sup> was 97.9% similar to <italic>Kineococcus radiotolerans</italic> and falls within the <italic>Kineococcus</italic> clade. Low 16S rRNA gene sequence similarities (&lt;94%) with other genera in the family <italic>Kineosporiaceae</italic>, including <italic>Angustibacter</italic> (93%), <italic>Kineosporia</italic> (94% to 95%), <italic>Pseudokineococcus</italic> (93%), <italic>Quadrisphaera</italic> (93%), and <italic>Thalassiella</italic> (94%) demonstrated that the strain B12<sup>T</sup> does not belong to these genera. Phylogenetic analysis of the <italic>gyrB</italic> gene show that all known <italic>Kineococcus</italic> species exhibited &lt;86% sequence similarity with B12<sup>T</sup>. Multi-locus sequence and whole genome sequence analyses confirmed that B12<sup>T</sup> clades with other <italic>Kineococcus</italic> species. Average nucleotide identity of strain B12<sup>T</sup> were 75–78% with other <italic>Kineococcus</italic> species, while values ranged from 72–75% with species from other genera within family <italic>Kineosporiaceae</italic>. Average amino-acid identities were 66–72% with other <italic>Kineococcus</italic> species, while they ranged from 50–58% with species from other genera. The dDDH comparison of strain B12<sup>T</sup> genome with members of genera <italic>Kineococcus</italic> showed 20–22% similarity, again demonstrating that B12<sup>T</sup> is distantly related to other members of the genus. Furthermore, analysis of whole proteome deduced from WGS places strain B12<sup>T</sup> in order <italic>Kineosporiales</italic>, confirming that strain B12<sup>T</sup> is a novel member of family <italic>Kineosporiaceae</italic>. Based on these analyses and other genome characteristics, strain B12<sup>T</sup> is assigned to a novel species within the genus <italic>Kineococcus</italic>, and the name <italic>Kineococcus rubinsiae</italic> sp. nov., is proposed. The type strain is B12<sup>T</sup> (=FJII-L1-CM-PAB2<sup>T</sup>; NRRL B-65556<sup>T</sup>, DSM 110506<sup>T</sup>).</p>