A Gram-reaction-positive, endospore-forming bacterium, designated strain P1T, was isolated from water samples collected from Pasinler Hot Spring and characterized using a polyphasic approach to clarify its taxonomic position. Strain P1T was found to have chemotaxonomic and morphological characteristics consistent with its classification in the genus Bacillus . The strain shared the highest 16S rRNA gene sequence identity values with Bacillus thermolactis R-6488T (97.6 %) and Bacillus kokeshiiformis MO-04T (97.2 %) and formed a distinct clade with both type strains in the phylogenetic trees based on 16S rRNA gene sequences. Strain P1T could grow optimally at 55 °C and in the presence of 2 % NaCl. The organism was found to contain meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The predominant menaquinone was determined to be MK-7. The major cellular fatty acids were identified as iso-C15 : 0, iso-C17 : 0 and anteiso-C17 : 0. Based upon the consensus of phenotypic and phylogenetic analyses, strain P1T represents a novel species of the genus Bacillus , for which the name Bacillus pasinlerensis sp. nov. is proposed. The type strain is P1T (=DSM 107529T=CECT 9885T=NCCB 100674T).

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Enzymes which would be active in cold conditions can be used in a wide range of fields from molecular biology to detergent industry due to their low processing capacity and high activity. In this study, sixty cold-adapted bacteria were isolated from water and sludge samples collected from Erzurum and Van provinces. Identification of eight isolates by molecular [(GTG) 5 -PCR and 16S rRNA sequencing] techniques and tests for temperature (4-35°C), pH (3-11) and salt (2-15% (w/v) requirements were performed. These bacteria were belonging to Pseudomonas chlororaphis subsp. aureofaciens (SM 01 1 A ), Psychrobacter faecalis (SM 01 2 D ), Rahnella aquatilis (SM 01 5 A ), Shewanella putrefaciens (SM 01 8 A ), Pseudomonas lactis (SM 01 10 A ), Flavobacterium chryseum (SM 01 12 E ), Exiguobacterium mexicanum (SM 01 17 A ) and Glutamicibacter arilaitensis (SM 01 18 A). The physicochemical requirements for all isolates ranged between 4-25°C, pH 5-7 and 2-15% salt (NaCl) concentration. However, E. mexicanum did not require salt in growth medium. All bacteria were evaluated for protease, lipase and amylase enzymes and all were found to be multiple enzyme producers. The eight isolates were identified from the resources of Turkey, for the first time and enzyme production abilities of some isolates to produce enzymes were declared. The originating of the producers of these enzymes from Turkey shows that Turkey has a remarkable reservoir for cold-adaptive microorganisms and these microorganisms will make important contributions to the detergent industry worldwide.

A novel Gram-stain-positive, rod-shaped, endospore-forming, motile, aerobic bacterium, designated as P2T, was isolated from a hot spring water sample collected from Ilica-Erzurum, Turkey. Phylogenetic analyses based on 16S rRNA gene sequence comparisons affiliated strain P2T with the genus Bacillus , and the strain showed the highest sequence identity to Bacillus azotoformans NBRC 15712T (96.7 %). However, the pairwise sequence comparisons of the 16S rRNA genes revealed that strain P2T shared only 94.7 % sequence identity with Bacillus subtilis subsp. subtilis NCIB 3610T, indicating that strain P2T might not be a member of the genus Bacillus . The digital DNA–DNA hybridization and average nucleotide identity values between strain P2T and B. azotoformans NBRC 15712T were 19.8 and 74.2 %, respectively. The cell-wall peptidoglycan of strain P2T contained meso-diaminopimelic acid. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an aminophospholipid, five unidentified phospholipids and two unidentified lipids while the predominant isoprenoid quinone was MK-7. The major fatty acids were iso-C15 : 0 and iso-C16 : 0. The draft genome of strain P2T was composed of 82 contigs and found to be 3.5 Mb with 36.1 mol% G+C content. The results of phylogenomic and phenotypic analyses revealed that strain P2T represents a novel genus in the family Bacillaceae , for which the name Calidifontibacillus erzurumensis gen. nov., sp. nov. is proposed. The type strain of Calidifontibacillus erzurumensis is P2T (=CECT 9886T=DSM 107530T=NCCB 100675T). Based on the results of the present study, it is also suggested that Bacillus azotoformans and Bacillus oryziterrae should be transferred to this novel genus as Calidifontibacillus azotoformans comb. nov. and Calidifontibacillus oryziterrae comb. nov., respectively.

In this study, one hundred and thirty isolates were isolated from water and sludge samples taken from hot springs located in different regions of Turkey. Among them, eleven isolates were chosen according to conventional (morphological, physiological and biochemical tests) and molecular methods (rep-PCR and 16S rRNA sequencing). These bacteria were then tested for their capability to produce valuable enzymes. As a result; species belonging to Bacillus, Anoxybacillus, Aeribacillus, Enterococcus, Exiguobacterium and Paenibacillus were identified. Test strains were found to have optimum reproductive potential at pH 5.0-9.0 and 15-65°C, usually at a concentration of 1.0-10.0% (w/v) NaCl. In addition, all thermotolerant bacteria were Gram, endospore (except E. profundum), catalase and oxidase (except E. faecium and E. profundum) positive, and rod-shaped (except E. faecium). It was observed that all isolates had a 99% similarity percentage as a result of 16S rRNA sequence analysis. All of the isolates were capable of producing industrially important enzymes moreover, eight of them could produce at least two of these enzymes. Test strains had high potential of industrial enzyme production, and the enzymes from these thermo-tolerant isolates will be widely used in biotechnological processes.