In this study, the isolation, identification and characterization of the thermophilic bacteria from different hot springs in Turkey were carried out by conventional (morphological, physiological and biochemical tests) and molecular methods (fatty acid methyl esters, GTG5-PCR and 16S rRNA sequencing). These thermophilic bacteria were then tested for their capability to produce enzymes such as lipase, protease, amylase and cellulase. O20 strain is a novel species according to identification studies. All of its isolates were capable of producing industrially valuable enzymes based on screening. In fact, most of them could produce at least two of these enzymes.

An extracellular thermostable alkaline serine protease enzyme from Aeribacillus pallidus C10 (GenBank No: KC333049), was purified 4.85 and 17. 32-fold with a yield of 26.9 and 19.56%, respectively, through DE52 anion exchange and Probond affinity chromatography. The molecular mass of the enzyme was determined through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), with approximately 38.35 kDa. The enzyme exhibited optimum activity at pH 9 and at temperature 60 °C. It was determined that the enzyme had remained stable at the range of pH 7.0–10.0, and that it had preserved more than 80% of its activity at a broad temperature range (20–80 °C). The enzyme activity was found to retain more than 70% and 55% in the presence of organic solvents and commercial detergents, respectively. In addition, it was observed that the enzyme activity had increased in the presence of 5% SDS. K_M and V_max values were calculated as 0.197 mg/mL and 7.29 μmol.mL⁻¹.min⁻¹, respectively. Graphical Abstract

Thermo alkaline mannanase was purified from the bacteria of Bacillus pumilus (M27) using the techniques of ammonium sulphate precipitation, DEAE-Sephadex ion exchange chromatography and Sephacryl S200 gel filtration chromatography with 111-fold and 36 % yield. It was determined that the enzyme had 2 sub-units including 35 kDa and 55 kDa in gel filtration chromatography and SDS-PAGE electrophoresis systems. The optimum pH and temperature was determined as 8 and 60 °C, respectively. It was also noticed that the enzyme did not lose its activity at a wide interval such as pH 3–11 and at high temperatures such as 90 °C. Additionally, the effects of some metal ions on the mannanase enzyme activity. Moreover, the clarifying efficiency of purified mannanase enzyme with some fruit juices such as orange, apricot, grape and apple was also investigated. Enzymatic treatment was carried out with 1 mL L⁻¹ of purified mannanase for 1 h at 60 °C. It was determined that the highest pure enzyme was efficient upon clarifying the apple juice at 154 % rate.

A Gram-reaction-positive, endospore-forming bacterium, designated strain P1^T, was isolated from water samples collected from Pasinler Hot Spring and characterized using a polyphasic approach to clarify its taxonomic position. Strain P1^T 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-6488^T (97.6 %) and Bacillus kokeshiiformis MO-04^T (97.2 %) and formed a distinct clade with both type strains in the phylogenetic trees based on 16S rRNA gene sequences. Strain P1^T 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-C_15 : 0, iso-C_17 : 0 and anteiso-C_17 : 0. Based upon the consensus of phenotypic and phylogenetic analyses, strain P1^T represents a novel species of the genus Bacillus , for which the name Bacillus pasinlerensis sp. nov. is proposed. The type strain is P1^T (=DSM 107529^T=CECT 9885^T=NCCB 100674^T).