Hot Spring In Turkey Research Articles

Isolation and Molecular Identification of Xylanase and Glucose-Isomerase Producer Geobacillus and Brevibacillus Strains from Hot Springs in Turkey

Isolation and Molecular Identification of Xylanase and Glucose-Isomerase Producer Geobacillus and Brevibacillus Strains from Hot Springs in Turkey

Purification and characterization of novel α-amylase from Anoxybacillus ayderensis FMB1

The thermophilic amylase producing Anoxybacillus ayderensis FMB1 was isolated from Sorgun Hot Spring in Turkey. In order to obtain improved production of the enzyme for cost-effective process, optimization of the process parameters is required. Optimum conditions for amylase production were determined using the traditional Submerged Fermentation (SmF) method. The maximum amylase activity was 295.8 U/mg, under the optimized conditions (24-h, 55 °C, and pH 8.0) in liquid BMB medium. The best enzyme production was obtained in the presence of 1.5% soluble starch as carbon source and 1% yeast extract as nitrogen source. Enzyme was produced under optimal conditions and partially purified. Partially purified enzyme was stable for up to 120 minutes at 50 °C, and its stability was found to be pH 5.0–10.0. Regarding the effects of some metals and chemicals on the activity of the partially purified enzyme, Ca2+ increased the activity by 21%, but enzyme activity was decreased by Mg2+, Zn2+, Ni2+, and Cu2+ by 5%, 29%, 57%, and 64%, respectively. and the enzyme activity was completely inhibited by Hg2+. In addition; DTT, PMSF, PCMB, SDS and EDTA caused a decrease in enzyme activity by 3%, 4%, 12%, 27% and 77%, respectively. The molecular weight of enzyme was found to be 58.5 kDa by SDS-PAGE. The results obtained show that the thermophilic A. ayderensis FMB1 and their enzyme have biotechnological importance.

Genome-based classification of Calidifontibacillus erzurumensis gen. nov., sp. nov., isolated from a hot spring in Turkey, with reclassification of Bacillus azotoformans as Calidifontibacillus azotoformans comb. nov. and Bacillus oryziterrae as Calidifontibacillus oryziterrae comb. nov.

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.

Whole-Genome Sequence of Geobacillus thermoleovorans ARTRW1, Isolated from Armutlu Geothermal Spring, Turkey.

The thermophilic microorganism Geobacillus thermoleovorans ARTRW1 was isolated from water samples collected in the Armutlu hot spring in Turkey. Here, the whole-genome sequence and its annotations are reported.

Bacillus pasinlerensis sp. nov., a thermophilic bacterium isolated from a hot spring in Turkey.

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).

Characterization and biotechnological application of protease from thermophilic Thermomonas haemolytica.

In this study, it was aimed to determine the ability to produce protease enzyme of Thermomonas haemolytica isolated from geothermal Nenehatun hot spring in Turkey and utilization of this enzyme in the detergent industry to remove protein stains. The protease-producing strains were screened from hot springs, and a potential strain was identified as T. haemolytica according to morphological, physiological and biochemical characteristics and sequence of 16S rRNA gene. Maximum protease activity was observed at 55°C and pH 9.0 at 72h of incubation. Activity was very stable between 50 and 65°C and pH 8.0-10.0, respectively. The enzyme activity was significantly inhibited by PMSF and partly inhibited by EDTA, EGTA, SDS, and urea. Some divalent metal ions such as Ca2+, Mg2+, and Mn2+ increased the enzyme activity, while Zn2+ and Cu2+ decreased. Michaelis-Menten constant (Km) and maximum velocity (Vmax) values were calculated by Lineweaver-Burk plot as 125EU/ml and 1262mg/ml, respectively. The biochemical characterization of the protease obtained from T. haemolytica was performed and applied on the blood and grass-stained fabrics with detergent to evaluate the stain removal performance of the enzyme. It was observed that the application of detergent with enzyme was more effective than the detergent without enzyme to clean up the stained fabrics. This is the first report of characterization of the protease of T. haemolytica. According to results obtained from this study, this new strain is a promising candidate for industrial applications in production of detergent.

Screening of Xylanase and Glucose Isomerase Producing Bacteria Isolated from Hot Springs in Turkey

The aim of this study was screening of xylanase and glucose isomerase producing thermophilic bacteria isolated from some hot springs located to Aegean Region of Turkey. Total sixty eight thermophilic isolates (Anoxybacillus, Brevibacillus, Geobacillus, Aneurinibacillus, Thermus, Paenibacillus and Proteobacter) were collected previously from these fields and identified based on 16S rDNA gene sequences. The isolates were screened with plate assay for determining the xylanase and glucose isomerase production abilities seperately in order to find new strains for industrial processes. After an incubation period of two days for xylanase and 5-6 days for glucose isomerase at 50-60 °C, positive strains were determined. Enzyme producing strains were confirmed by spectrophotemetric measurements with crude enzyme extracts, birchwood xylan and glucose were used as substrates. Most of the strains (59 strains) were positive for xylan degradation while only sixteen of the strains had showed glucose isomerase activity. Fourteen of the strains have showed both xylanase and glucose isomerase activity. None of the Paenibacillus, Aneurinibacillus, and Proteobacter strains were glucose isomerase positive, although the glucose isomerase activity of Geobacillus strains were notably high. Both xylanase and glucose isomerase activities have observed at 50-60 °C which is suitable for biotechnological applications.

Isolation and Characterization of Thermophilic Bacteria from Geothermal Areas in Turkey and Preliminary Research on Biotechnologically Important Enzyme Production

ABSTRACTIn 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.

Complete genome of the potential thermozyme producer Anoxybacillus gonensis G2T isolated from the Gönen hot springs in Turkey

Anoxybacillus gonensis type strain G2T (=NCIMB 13,933T =NCCB 100040T) has been isolated from the Gönen hot springs in Turkey. This strain produces a number of well-studied, biotechnologically important enzymes, including xylose isomerase, carboxylesterase, and fructose-1,6-bisphosphate aldolase. In addition, this strain is an excellent candidate for the bioremediation of areas with heavy metal pollution. Here, we present a high-quality, annotated, complete genome of A. gonensis G2T. Furthermore, this report provides insights into several novel enzymes of strain G2T and their potential industrial applications.

Anoxybacillus sp. AH1, an α-amylase-producing thermophilic bacterium isolated from Dargeçit hot spring

AbstractThe present study was conducted to isolate α-amylase-producing thermophilic bacteria from Darge¸cit hot springs in Turkey. The morphological, biochemical and physiological characterisation, as well as genetic analysis by 16S rRNA sequences indicated that the isolated strain AH1 was a member of Anoxybacillus genus. The strain was aerobe, Gram-positive and spore-forming rod, exhibiting optimum growth temperature and pH of 60ºC and 7.0-7.5, respectively. Optimization of growth medium and enzyme assay conditions for extracellular α-amylase production by the novel thermophilic Anoxybacillus sp. AH1 were carried out in many different media containing a variety of carbon and nitrogen sources. Among various carbon and nitrogen sources, peptone (2054.1 U/mL) at 1% and maltose (1862.9 U/mL) at 0.5% increased α-amylase activity, compared to controls. Moreover, a high enzyme production was observed with potato starch at 0.5% and 1% (2668.4 U/mL and 3627 U/mL, respectively), as well as with 1% soluble starch (2051.9 U/mL). The enzyme activity was found to be rather high in the presence of CaCl

Heat shock protein 70 purification and characterization from Cyprinion macrastomus macrastomus and Garra rufa obtusa

Organisms resist stress factors by common mechanisms at cellular and physiological levels. Heat shock proteins (Hsps) play an essential role as a protective mechanism against stressors, and Hsp70 has a central role in the heat shock response. Although several investigations have been made on the cellular functions of Hsps, very little is known about extreme temperature effects on organisms like fish. To address this point, we have studied two fish species, Cyprinion macrostomus macrostomus and Garra rufa obtuse, whose habitat is the Kangal hot spring in Turkey. The hot spring has multiple stress factors; extremely hot temperature, food deprivation, infectious medium and low levels of oxygen. Like other Hsp70s, two Cyprinidae species Hsp70 showed similar ATPase and substrate protein folding activities in vitro. These proteins also showed relatively higher thermal stability. Biochemical characterization of two Hsp70, suggested a Hsp mechanism capable of protecting the cell against stressors even under adverse conditions and in the presence of multiple stress factors.

Cloning and Sequence Analysis of Novel DNA Polymerases from Thermophilic Geobacillus Species Isolated from Hot Springs in Turkey: Characterization of a DNA Polymerase I from Geobacillus kaue Strain NB

The complete coding sequences of the polA genes from seven thermophilic Geobacillus species, isolated from hot springs of Gönen and Hisaralan in Turkey, were cloned and sequenced. The polA genes of these Geobacillus species contain a long open reading frame of 2,637 bp encoding DNA polymerase I with a calculated molecular mass of 99 kDa. Amino acid sequences of these Geobacillus DNA polymerases are closely related. The multiple sequence alignments show all include the conserved amino acids in the polymerase and 5'-3' exonuclease domains, but the catalytic residues varied in 3'-5' exonuclease domain of these Geobacillus DNA polymerases. One of them, DNA polymerase I from Geobacillus kaue strain NB (Gkaue polI) is purified to homogeneity and biochemically characterized in vitro. The optimum temperature for enzymatic activity of Gkaue polI is 70 °C at pH 7.5-8.5 in the presence of 8 mM Mg(2+) and 80-100 mM of monovalent ions. The addition of polyamines stimulates the polymerization activity of the enzyme. Three-dimensional structure of Gkaue polI predicted using homology modeling confirmed the conservation of all the functionally important regions in the polymerase active site.

Screening of thermophilic bacteria producing glucose isomerase from some hot springs in Turkey

Screening of thermophilic bacteria producing glucose isomerase from some hot springs in Turkey

Screening of xylanolytic thermophilic bacteria from some hot springs in Turkey

Screening of xylanolytic thermophilic bacteria from some hot springs in Turkey

Isolation and characterization of extracellular protease producing Geobacillus sp. from various hot springs in Turkey

Isolation and characterization of extracellular protease producing Geobacillus sp. from various hot springs in Turkey

Isolation and characterization of xylanolytic new strains of Anoxybacillus from some hot springs in Turkey

Some hot springs located in the west of Turkey were investigated regarding the presence of xylanolytic thermophilic microorganisms. Based on phenotyping characteristics and 16S rRNA gene sequence analysis, 9 of these xylanolytic isolates belonged to the genus Anoxybacillus, and grew optimally at about 50-60 °C on nutrient agar. The 16S rRNA gene sequence analysis showed that these isolates resembled the Anoxybacillus species =>97 and these isolates are members of the genus Anoxybacillus. Based on the DNA-DNA hybridization study, SDS-PAGE profile, and biochemical and physiological features, I3, CT1Sari, and BT2.1 isolates are new strains of Anoxybacillus gonensis; I4.2 and B9.3 isolates are new strains of A. voinovskiensis; and I4.1, AC26, ACT14, and ACT2Sari isolates are new strains of A. kestanbolensis. The presence of xylanase activities, their optimum temperature, pH stability, and optimum pH were also investigated. The isolates ACT2Sari and ACT14 had the highest temperature optima (75 °C), and I3, CT1Sari, BT2.1, ACT2Sari, and ACT14 had the highest pH optima (pH 9.0) of xylanase. The xylanases of I3, CT1Sari, BT2.1, I4.1, ACT2Sari, AC26, and ACT14 were optimally active both at alkaline pH and elevated temperature, and xylanases of I4.1, ACT2Sari, AC26, and ACT14 also were stable at alkaline pH.

Identification and characterization of thermophilic bacteria isolated from hot springs in Turkey

The present study was conducted to identify and characterize the thermophilic bacteria isolated from various hot springs in Turkey by using phenotypic and genotypic methods including fatty acid methyl ester and rep-PCR profilings, and 16S rRNA sequencing. The data of fatty acid analysis showed the presence of 17 different fatty acids in 15 bacterial strains examined in this study. Six fatty acids, 15:0 iso, 15:0 anteiso, 16:0, 16:0 iso, 17:0 iso, and 17:0 anteiso, were present in all strains. The bacterial strains were classified into three phenotypic groups based on fatty acid profiles which were confirmed by genotypic methods such as 16S rRNA sequence analysis and rep-PCR genomic fingerprint profiles. After evaluating several primer sets targeting the repetitive DNA elements of REP, ERIC, BOX and (GTG) 5, the (GTG) 5 and BOXA1R primers were found to be the most reliable technique for identification and taxonomic characterization of thermophilic bacteria in the genera of Geobacillus, Anoxybacillus and Bacillus spp. Therefore, rep-PCR fingerprinting using the (GTG) 5 and BOXA1R primers can be considered as a promising genotypic tool for the identification and characterization of thermophilic bacteria from species to strain level.

Cloning, expression, purification and characterisation of a thermostable chitinase fromBacillus licheniformis A1

The chitinase B gene (chiB65) ofBacillus licheniformis A1 (BlicA1) isolated from Diyadin hotspring in Turkey was cloned and sequenced. The gene is 1779 bp long and encodes a protein 592 amino acids with a 35-amino acid signal peptide at N-terminal. The gene has 99% percent similarity tochiB gene ofBacillus licheniformis under the GenBank number AY205293. The gene without signal peptide was overexpressed inEscherichia coli and the recombinant protein purified by nickel affinity chromatography. The activity of enzyme was shown on SDS-PAGE with the flourogenic substrate 4-methylumbelliferyl β-D-N,N′-diacetylchitobioside. Kinetic characterisation of the enzyme was performed at 65 °C by using chromogenic substratep-nitrophenylN,N′-diacetyl-β-D-chitobioside, andK m and Vmax were found to be 0.02 μM and 1017 U/mg protein, respectively. Enzyme has maximal activity at pH 6.0 and was stable over a broad pH range of 5.0–9.0 for 24 h at room temperature and 4 h at 65 °C. Enzyme was 60% stable at 65 °C for 1.5 h. The inhibition or activation of some substances on the activity of enzyme was determined. High kinetic properties of enzyme open the possibility of an extensive structural and enzyme-substrate interaction studies.

Ichthyotherapy as alternative treatment for patients with psoriasis: a pilot study.

Ichthyotherapy (therapy with the so-called ‘Doctorfish of Kangal’, Garra rufa) has been shown to be effective in patients with psoriasis in the Kangal hot springs in Turkey. This study evaluates the efficacy and safety of ichthyotherapy in combination with short-term ultraviolet A sunbed radiation in the treatment of psoriasis under controlled conditions. We retrospectively analyzed 67 patients diagnosed with psoriasis who underwent 3 weeks of ichthyotherapy at an outpatient treatment facility in Lower Austria between 2002 and 2004. Main outcome measures are as follows: overall relative reduction in Psoriasis Area Severity Index (PASI) score; proportion of patients with an improvement in their PASI score of ≥75% (PASI-75) and ≥50% (PASI-50); patient-reported outcomes assessed with a custom questionnaire; and patient follow-up with a questionnaire sent out in March 2005. Safety was evaluated by reviewing adverse events and vital signs. Overall there was a 71.7% reduction in PASI score compared to baseline (P < 0.0001). Of the 67 patients studied, 31 (46.3%) achieved PASI-75 and 61 patients (91%) achieved at least PASI-50. Patients reported substantial satisfaction with the treatment. The reported mean remission period was 8.58 months [95% confidence interval (CI) 6.05–11.11]. A total of 87.5% of patients reported a more favorable outcome with ichthyotherapy, when asked to compare ichthyotherapy to other previously tried therapies. Sixty-five percent stated that after the relapse their symptoms were less severe than before treatment. There were no significant adverse events. The benefit demonstrated in this study along with the favorable safety profile suggests that ichthyotherapy could provide a viable treatment option for patients with psoriasis.

Balneotherapy in dermatology.

Balneotherapy and spa therapy emerged as an important treatment modality in the 1800s, first in Europe and then in the United States. Balneotherapy involves immersion of the patient in mineral water baths or pools. Today, water therapy is being practiced in many countries. Examples of unique and special places for balneotherapy are the Dead Sea in Israel, the Kangal hot spring in Turkey, and the Blue Lagoon in Iceland. Bathing in water with a high salt concentration is safe, effective, and pleasant for healing and recovery. This approach needs no chemicals or potentially harmful drugs. There are almost no side effects during and after treatment, and there is a very low risk to the patient's general health and well-being. Mineral waters and muds are commonly used for the treatment of various dermatologic conditions. The major dermatologic diseases that are frequently treated by balneotherapy with a high rate of success are psoriasis and atopic dermatitis. The mechanisms by which broad spectrums of diseases are alleviated by spa therapy have not been fully elucidated. They probably incorporate chemical, thermal, mechanical, and immunomodulatory effects. The major importance of balneotherapy and spa therapy both individually and as complements to other therapies lies in their potential effectiveness after standard medical treatments have failed to give comfort to these patients.