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

A Gram-positive, coccoid, motile, aerobic bacterium, designated strain B12(T)was isolated from a Jet Propulsion Laboratory spacecraft assembly cleanroom, Pasadena, CA, United States. Strain B12(T)was resistant to chloramphenicol (100 mu g/mL), and is a relatively slow grower (3-5 days optimal). Strain B12(T)was found to grow optimally at 28 to 32 degrees C, pH 7 to 8, and 0.5% NaCl. Fatty acid methyl ester analysis showed that the major fatty acid of the strain B12(T)was anteiso C-15:0(66.3%), which is also produced by otherKineococcusspecies. However, arachidonic acid (C-20:4 omega 6,9,12,16c) was present in strain B12(T)andKineococcus glutinatusYIM 75677(T)but absent in all otherKineococcusspecies. 16S rRNA analysis revealed that strain B12(T)was 97.9% similar toKineococcus radiotoleransand falls within theKineococcusclade. Low 16S rRNA gene sequence similarities (<94%) with other genera in the familyKineosporiaceae, includingAngustibacter(93%),Kineosporia(94% to 95%),Pseudokineococcus(93%),Quadrisphaera(93%), andThalassiella(94%) demonstrated that the strain B12(T)does not belong to these genera. Phylogenetic analysis of thegyrBgene show that all knownKineococcusspecies exhibited Kineococcusspecies. Average nucleotide identity of strain B12(T)were 75-78% with otherKineococcusspecies, while values ranged from 72-75% with species from other genera within familyKineosporiaceae. Average amino-acid identities were 66-72% with otherKineococcusspecies, while they ranged from 50-58% with species from other genera. The dDDH comparison of strain B12(T)genome with members of generaKineococcusshowed 20-22% similarity, again demonstrating that B12(T)is distantly related to other members of the genus. Furthermore, analysis of whole proteome deduced from WGS places strain B12(T)in orderKineosporiales, confirming that strain B12(T)is a novel member of familyKineosporiaceae. Based on these analyses and other genome characteristics, strain B12(T)is assigned to a novel species within the genusKineococcus, and the nameKineococcus rubinsiaesp. nov., is proposed. The type strain is B12(T)(=FJII-L1-CM-PAB2(T); NRRL B-65556(T), DSM 110506(T)).