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Full 16S rDNA sequence-based phylogenetic neighbour-joining tree showing the phylogenetic relationship of strain AH1 relative to other strains of the genus Anoxybacillus. Aeribacillus pallidus DSM 3670 is used as the out-group. Bar indicates 0.1 nucleotide substitutions per position.
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The 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-positi...
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... isolate was identified using the EzTaxon server (http://www.ezbiocloud.net/eztaxon; Kim et al. 2012) on the basis of 16S rRNA sequence data. The re- sults of 16S rRNA gene sequence analysis showed that the strain AH1 is most similar (98.23% similarity) to Anoxybacillus flavithermus subsp. flavithermus DSM 2641 T (GenBank accession No.: Z26932). The strain 858¨O 858¨ 858¨O. Acer et al. . The presentation of results as a phylogenetic tree showed that the isolate was separated from other species within the Bacillus radiation, and from the phylogenetically related Anoxy- bacillus species (Fig. ...
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... U/mL) and in the presence of 2% (w/v) soluble starch (10837.7 U/mL). The enzyme activity was enhanced in the presence of 1.5 mM Mn 2+ (123%), while it was strongly inhibited 1.5 mM by Hg 2+ (Bekler and Guven 2014;Acer et al. 2015) conducted isolation of α-amylase producing thermophilic strain AH1 which belongs to Anoxybacillus genus from Dargecit hot springs in Turkey. As a result of numerous carbon and nitrogen sources tested, peptone (2054.1 U/mL) at 1% and maltose (1862.9 ...
The Bacillaceae family members are considered to be a good source of microbial factories for biotechnological processes. In contrast to Bacillus and Geobacillus, Anoxybacillus, which would be thermophilic and spore-forming group of bacteria, is a relatively new genus firstly proposed in the year of 2000. The development of thermostable microbial enzymes, waste management and bioremediation processes would be a crucial parameter in the industrial sectors. There has been increasing interest in Anoxybacillus strains for biotechnological applications. Therefore, various Anoxybacillus strains isolated from different habitats have been explored and identified for biotechnological and industrial purposes such as enzyme production, bioremediation and biodegradation of toxic compounds. Certain strains have ability to produce exopolysaccharides possessing biological activities including antimicrobial, antioxidant and anticancer. This current review provides past and recent discoveries regarding Anoxybacillus strains and their potential biotechnological applications in enzyme industry, environmental processes and medicine.
... High amount of amylase production was observed also for Anoxybacillus sp. [26] and A. flavithermus SO-13 [27] with potato starch. The only Brachybacterium strain reported in the literature B. paraconglomeratum Strain LB 25 was studied with potato starch [14]. ...
... In another study, the highest amylase production from Anoxybacillus sp. AH1 was observed with maltose supplementation, glucose, and lactose were also increased the enzyme production [26]. The inhibitory effect of glucose on FAD4 was also observed for Anoxybacillus flavithermus. ...
A novel amylase producing bacterium FAD4 was isolated from the wastewater of a textile factory located in Soke (Aydın/Turkey). The amylase production ability of gram positive, coccoidal FAD4 strain was confirmed with plate assay. Morphological and 16S rRNA sequence analyses revealed that FAD4 belongs to the Brachybacterium paraconglomeratum species with a sequence similarity of 99.8%. The optimal conditions for amylase production were determined as 72 h at 30 °C with supplementation of 1% starch. Optimum temperature and pH of the amylase were 50 °C and 7.0 respectively. Different starch, carbon and nitrogen sources were investigated for amylase production. A high enzyme production was observed with 1% potato starch and among nitrogen sources peptone was induced the production of amylase. Lactose, galactose, and fructose were also increased the enzyme production as carbon sources.
... Hot springs in eastern and southeastern Turkey are reservoirs for natural resources that are attractive for biotechnological studies (Guven et al. 2018). Various thermophilic microbial species have been reported in many studies carried out in the Hasanabdal thermal springs (Poli et al. 2012;Savas et al. 2009;Yanmis et al. 2015), Sirnak (Derya and Ahmet 2014), Diyadin (Baltaci et al. 2017), Dargecit (Acer et al. 2015), Guclukonak (Poli et al. 2012) and Pasinler (Adiguzel et al. 2011;Oztas Gulmus and Gormez 2020b). The presence of many thermophilic Bacillus and Geobacillus species has been reported in the Pasinler and Ilica thermal spring of Erzurum by different researchers (Oztas Gulmus and Gormez 2020b;Yilmaz et al. 2016). ...
In the last two decades, researchers have increasingly focused on the rich microorganism-based diversity of natural hot spring sources to explore the benefits of thermophiles in industrial and biotechnological fields. Within the scope of this study, a total of 83 thermophilic Bacilli strains were isolated from 7 different geothermal hot springs (at temperatures ranging between 40 and 85 °C) located in the Eastern and Southeastern Anatolia Regions of Turkey. The physiological, morphological, biochemical and molecular properties of the isolates were determined. As a result of the 16S rRNA gene sequence analysis, 5 different species (Bacillus licheniformis, Bacillus sp., Bacillus subtilis, Geobacillus kaustophilus, and Weizmannia coagulans,) were identified. B. licheniformis and B. subtilis were the most frequently encountered species among those obtained from the researched hot spring sources. Phylogenetic analysis was conducted to evaluate the phylogenetic relationships of the isolated species. The results showed that there was no significant difference between the groups and the bacteria in terms of the locations or optimum temperatures of the isolates. The bacterial isolates were screened for amylase, cellulase, lipase and protease hydrolytic enzyme activities. The hydrolytic enzyme production potentials among the isolates were identified in 68 (82%) isolates for amylase, 34 (41%) for cellulase, 69 (83%) for lipase and 73 (88%) for protease. All isolates were found to have at least one or more extracellular enzyme activities. Additionally, it was determined that 27 of the existing isolates (32.8%) were able to produce all of the aforementioned hydrolytic enzymes.
... It was observed Anoxybacillus rupiensis performed well at neutral pH. Acer et al., (2015) also reported the similar results with maximum growth at pH 7 by the same specie Anoxybacillus but Jabeen et al. (2018) found pH 6 the most appropriate for the Anoxybacillus beppuensis. Nitrogen source is an important factor influenced bacterial growth and protein production. ...
... Yeast extracts and Peptone were found to be growth enhancer for both strains JF83 and JF82, respectively. Bakir and Metin (2015) and Acer et al. (2015) also found peptone the most suitable for the production of enzyme. ...
... Effect of different nitrogen sources on protein production was performed in order to get suitable nitrogen source. Acer et al. [19] also found peptone which is also an organic nitrogen source most suitable for the production of enzyme/protein from Anoxybacillus. ...
Extremophiles are the organisms that can be manipulated for various industrial activities. Agro–dairy wastes are being utilized for economical production of enzymes employing microbes. In this regard, thermophilic amylolytic bacterium was isolated from soil in vicinity of hot spring near Chakwal, Pakistan. The isolated bacterium was rod‐shaped, positive for Gram staining, and endospores former. The isolate was able to utilize sugars like, maltose, xylose, fructose, sachorose, sorbitol, mannose, mannitol, arabinose and polysaccharides like gelatin and starch. The isolate grew over a wide temperature range (40–70°C) and pH (4–9). It exhibited enough growth with different nitrogen sources which could produce amylase up to 0.72 U/mL, while hydrolyzing starch. Phylogenetic analysis based on 16S rRNA gene sequence showed that it belongs to Anoxybacillus beppuensis under Accession number KF254912 (JF84). The DNA G+C content were 56.59%. The isolate JF84 showed fourfold purification and percentage yield was estimated to be 94.21% as compared to crude enzyme. The decreasing order of substrate suitability for amylase production was found to be sugarcane bagasse (SCB) > wheat straw (WS) > sugarcane bagasse and whey (SB + W) > whey (W). A. beppuensis JF84 yielded enough thermostable α‐amylase in economical substrates as well as tolerated broad pH and temperature suggesting its wide economical commercial use in amylase‐dependent industries. © 2018 American Institute of Chemical Engineers Environ Prog, 38: 417–423, 2019
... In another study, Acer et al. (2015) isolated α-amylase-producing thermophilic bacteria from the mud of Dargecit hot spring (water temperature and pH as 58 °C and 6.9, respectively) in Mardin Province of Turkey. The isolated strain AH1 was found to be a member of Anoxybacillus genus by characterising with the morphological, biochemical and physiological tests, in addition to the genetic analysis by 16S rRNA sequences. ...
Turkey has a great number of different ecological areas, owning over 200 hot water resources and various hypersaline environments with a broad microbial diversity and opportunities for newly isolated microorganisms from extreme environments for many industrial applications. A variety of thermophilic and halophilic microorganisms in different regions of Turkey have been isolated and identified. The thermophilic bacterial members studied were Anoxybacillus, Geobacillus, Bacillus, Brevibacillus, and Aeribacillus belonging to the Bacillaceae family and the other thermophiles such as Thermus and Thermomonas, whereas the isolated halophilic microorganisms were mainly found to be members of the archaeal family Halobacteriaceae or grouped into bacterial phylum Bacteroidetes. In summary, the present study reviews on (1) isolating and identifying thermophiles and halophiles single or as community from various extreme habitats in Turkey based on conventional (morphological, physiological and biochemical tests) and/or molecular methods, (2) screening these extremophiles for industrially important enzymes, (3) studying other novel products and their use in other areas of biotechnology, and finally (4) discussing about the development strategies and the future perspectives on poorly studied extremophilic microorganisms in the country to fulfill future biotechnological and industrial demands.
... In another study, Acer et al. (2015) isolated α-amylase-producing thermophilic bacteria from the mud of Dargecit hot spring (water temperature and pH as 58 °C and 6.9, respectively) in Mardin Province of Turkey. The isolated strain AH1 was found to be a member of Anoxybacillus genus by characterising with the morphological, biochemical and physiological tests, in addition to the genetic analysis by 16S rRNA sequences. ...
Turkey has a great number of different ecological areas, owning over two hundred hot water resources and various hypersaline environments with a broad microbial diversity and opportunities for newly isolated microorganisms from extreme environments for many industrial applications. A variety of thermophilic and halophilic microorganisms in different regions of Turkey have been isolated and identified. The thermophilic bacterial members studied were Anoxybacillus, Geobacillus, Bacillus, Brevibacillus, Aeribacillus belonging to the Bacillaceae family, and the other thermophiles such as Thermus and Thermomonas, whereas the isolated halophilic microorganisms were mainly found to be members of the archaeal family Halobacteriaceae or grouped into bacterial phylum Bacteroidetes.
In summary, the present study reviews on the (i) isolation and identification of thermophiles and halophiles single or as community from various extreme habitats in Turkey based on conventional (morphological, physiological and biochemical tests) and/or molecular methods (ii) screening these extremophiles for industrially important enzymes and (iii) studying other novel products and their use in other areas of biotechnology (iv) and finally discussing about the development strategies and the future perspectives on poorly–studied extremophilic microorganisms in the country to fulfill future biotechnological and industrial demands.
... Many thermostable α-amylases from the genera Anoxybacillus and Geobacillus were characterized including: Anoxybacillus flavithermus (Bolton et al, 1997;Tawil et al. 2012;Agüloğlu Fincan et al. 2014;Ozdemir et al. 2015Ozdemir et al. , 2016a, Anoxybacillus amylolyticus (Poli et al. 2006) and Anoxybacillus caldiproteolyticus D504 and D621 (Ozdemir et al. 2016b), Anoxybacillus spp. KP1, SK3-4 (ASKA), DT3-1 (ADTA), TSSC-1, IB-A, AH1, and YIM 342 (Chai et al. 2012;Kikani and Singh 2012;Hauli et al. 2013;Güven 2014;Acer et al. 2015;Zhang et al. 2015) in addition to Geobacillus thermoleovorans MTCC 4220 (Gt-amyII), CCB_US3_UF5 (GTA ), YN, NP54 (Berekaa et al. 2007; Rao and Satyanarayana 2007;Mok et al. 2013;Mehta and Satyanarayana 2014), G. thermoleovorans subsp. stromboliensis Pizzo (amyA), (Finore et al. 2011), Geobacillus thermodenitrificans HRO10 (Ezeji and Bahl 2006), and Geobacillus sp. ...
Among the thermophilic Bacillaceae family members, α-amylase production of 15 bacilli from genus Anoxybacillus was investigated, some of which are biotechnologically important. These Anoxybacillus α-amylase genes displayed ≥ 91.0% sequence similarities to Anoxybacillus enzymes (ASKA, ADTA and GSX-BL), but relatively lower similarities to Geobacillus (≤ 69.4% to GTA, Gt-amyII), and Bacillus aquimaris (≤ 61.3% to BaqA) amylases, all formerly proposed only in a Glycoside Hydrolase 13 (GH13) subfamily. The phylogenetic analyses of 63 bacilli-originated protein sequences among 93 α-amylases revealed the overall relationships within Bacillaceae amylolytic enzymes. All bacilli α-amylases formed 5 clades different from 15 predefined GH13 subfamilies. Their phylogenetic findings, taxonomic relationships, temperature requirements, and comparisonal structural analyses (including their CSR-I-VII regions, 12 sugar- and 4 calcium-binding sites, presence or absence of the complete catalytic machinery, and their currently unassigned status in a valid GH13 subfamiliy) revealed that these five GH13 α-amylase clades related to familly share some common characteristics, but also display differentiative features from each other and the preclassified ones. Based on these findings, we proposed to divide Bacillaceae related GH13 subfamilies into 5 individual groups: the novel a2 subfamily clustered around α-amylase B2M1-A (Anoxybacillus sp.), the a1, a3 and a4 subfamilies (including the representatives E184aa-A (Anoxybacillus sp.), ATA (Anoxybacillus tepidamans), and BaqA,) all of which were composed from the division of the previously grouped single subfamily around α-amylase BaqA, and the undefinite subfamily formerly defined as xy including Bacillus megaterium NL3.
Thermophilic and alkaliphilic microorganisms are unique organisms that possess remarkable survival strategies, enabling them to thrive on a diverse range of substrates. Anoxybacillus, a genus of thermophilic and alkaliphilic bacteria, encompasses 24 species and 2 subspecies. In recent years, extensive research has unveiled the diverse array of thermostable enzymes within this relatively new genus, holding significant potential for industrial and environmental applications. The biomass of Anoxybacillus has demonstrated promising results in bioremediation techniques, while the recently discovered metabolites have exhibited potential in medicinal experiments. This review aims to provide an overview of the key experimental findings related to the biotechnological applications utilizing bacteria from the Anoxybacillus genus.
α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60-80 °C and is active and stable over a wide pH range (4.0-9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.
