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Bacillus pakistanensis sp. nov., a halotolerant bacterium isolated from salt mines of the Karak Area in Pakistan


A rod shaped, non-motile, endospore forming, Gram-stain positive and moderately halotolerant strain, designated as NCCP-168T, was isolated from salt mines sampled in the Karak district of Khyber Pakhtunkhwa Province in Pakistan. To delineate its taxonomic position, the strain was subjected to polyphasic characterization. Cells of strain NCCP-168T can grow at 10–40 C (optimum at 30–35 C), in a pH range of 5.0–9.0 (optimum at pH 8.0) and in 0–17 % (w/v) NaCl on agar medium. The phylogenetic analysis based on the 16S rRNA gene sequence showed that strain NCCP-168T belongs to the genus Bacillus with the highest similarity to Bacillus seohaeanensis BH724T (97.1 %), and less than 97 % similarity with other closely related taxa (95.6 % with B. subtilis subsp. subtilis NCIB3610T). DNA–DNA relatedness between strain NCCP-168T and the type strains of closely related species was lower than 30 %. Chemotaxonomic data (major menaquinone, MK-7; cell wall peptidoglycan type, A1γ [meso-diaminopimelic acid]; major fatty acids, iso-C15:0 29.9 %, anteiso-C15:0 29.3 %, iso-C16:0 11.4 %, iso-C14:0 8.9 % and anteiso-C17:0 7.0 %; major polar lipids, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine) support the affiliation of strain NCCP-168T with genus Bacillus. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain NCCP-168T can be distinguished from the closely related taxa and thus represents a novel species in the genus Bacillus, for which the name Bacillus pakistanensis sp. nov. is proposed, with the type strain NCCP-168T (= KCTC 13786T = DSM 24834T = JCM 18975T).

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This work was partially supported by PSDP Project “Research for Agricultural Development Program (RADP)” under a sub-project (Grant No. CS-55/RADP/PARC) entitled “Establishment of Microbial Bio-Resource Laboratories: National Culture Collection of Pakistan (NCCP)” from Pakistan Agricultural Research Council (PARC), Islamabad, Pakistan. The partial financial support of KRIBB to authors, Aneela Roohi and Saira Abbas in the training course at Korean Collection for Type Cultures, Biological Resources Centre, KRIBB under the umbrella of ACM forum is also gratefully acknowledged.

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Correspondence to Iftikhar Ahmed or Young H. Chang.

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Aneela Roohi and Iftikhar Ahmed contributed equally to the experimentation.

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Roohi, A., Ahmed, I., Paek, J. et al. Bacillus pakistanensis sp. nov., a halotolerant bacterium isolated from salt mines of the Karak Area in Pakistan. Antonie van Leeuwenhoek 105, 1163–1172 (2014).

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  • Halotolerant bacterium
  • Karak salt mines
  • Bacillus pakistanensis