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Anoxybacillus sp AH1, an ?-amylase-producing thermophilic bacterium isolated from Dargecit hot spring
(Springer, 2015) Acer, Omer; Pirinccioglu, Hemse; Bekler, Fatma Matpan; Gul-Guven, Reyhan; Guven, Kemal
The present study was conducted to isolate alpha-amylase-producing thermophilic bacteria from Dargecit 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 60 degrees C and 7.0-7.5, respectively. Optimization of growth medium and enzyme assay conditions for extracellular a-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 a-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 CaCl2 up to 100 mM.
Bingöl Sıcak Su Kaplıcalarından İzole Edilen Bakterilerin Tanımlanması, Proteaz ve Amilaz Enzimlerinin Karakterizasyonu
(2021) Güven, Kemal; Bekler, Fatma Matpan; Aslan, Zindar
Bingöl İli Binkap sıcak su kaynaklarından izole edilen bakteriler karakterize edilerek, bunlara ait endüstriyel alanda önem arz eden proteaz ve amilaz gibi enzimleri üzerinde çalışmalar yapılmıştır. Bu çalışmada su ve toprak örneklerinden iki bakteri suşu izole edildi. Bu bakterilerin çeşitli biyokimyasal testleri ile morfolojik ve fizyolojik analizleri yapıldı. İzole edilen 4NK bakterisinin çubuk şeklinde Gram pozitif olduğu, 5NK bakterisinin de Gram pozitif ve çubuk şeklinde olduğu belirlenmiştir. İzole edilmiş bu iki bakteri için optimum pH değerinin 6.0 olduğu tespit edildi. Optimum üreme sıcaklıkları ise sırasıyla 40 ve 45 °C olarak belirlenmiştir. İzole edilen bakterilerin optimum koşulları belirlendikten sonra bunların ürettikleri enzimlerin optimum koşulları belirlendi. Buna göre 4NK bakterisinin proteaz ve amilaz enzimlerini üretim açısından optimum süreleri sırasıyla 18. saatte (195.80 U mg-1) ve 15. saatte (428.33 U mg-1) olduğu belirlenmiştir. Proteaz ve amilaz enzimlerinin optimum pH’sı 8.0, optimum sıcaklıkları sırasıyla 40-50 °C ve 50-60 °C olarak bulunmuştur. Ayrıca, 5NK bakteri varyetesi için proteaz ve amilaz enzimlerini üretim açısından optimum süresi, proteaz için 260.93 U mg-1ve amilaz için 380.58 U mg-1olmak üzere 24. saat olarak belirlenmiştir. Proteaz ve amilaz enzimlerinin optimum pH’ları 8.0, optimum sıcaklıkları 50 °C olarak bulunmuştur. İzolasyonu yapılan 4NK ve 5NK bakterilerinin 16S rRNA dizi analizi yapılmış, buna göre 4NK bakterisinin Bacillus subtilis türüne yakın, 5NK bakterisinin ise Bacillus paralicheniformis türüne filogenetik olarak yakın olduğu tespit edilmiştir.
Bingöl ve Diyarbakır İllerinde Yayılış Gösteren Bazı Dalgıç Böcek (Coleoptera: Dytiscidae) Türlerinin Mitokondrial Sitokrom Oksidaz Alt Ünite 1 Geni (COI) ile Filogenetik Analizi
(2021) Bekler, Fatma Matpan; Güven, Kemal; Uzen, Ramazan; Yıldırım, İbrahim Halil; Aykut, Medeni
Dytiscidae Leach, 1815 familyası sucul böceklerin önemli bileşenlerindendir. Familya üyeleri yırtıcı dalışböcekleri olarak bilinirler ve genellikle tüm sucul habitatlara uyum göstermişlerdir. Bu çalışmada, Bingöl veDiyarbakır illerinden Eylül 2016 ile Mayıs 2017 dönemlerinde toplanan Dytiscidae familyasının 7 cinsine ait 17türün PCR yöntemiyle mitokondriyal DNA örnekleri elde edilerek moleküler düzeyde araştırılmıştır. Bu türlerinmitokondriyal COI genlerinin nükleotid sekansları ve PCR genomik dizileri moleküler belirteç olarak kullanıldı.Mitokondriyal DNA dizi analizleri BLAST taraması yoluyla yapıldı. 17 türe ait mitokondriyal COI gen dizileri,türlerin tanımlamasında kullanılan CLC Sequence Viewer 8. programı yardımı ile filogenetik soy ağacıoluşturularak benzerlikleri karşılaştırıldı. Türlerin yakınlık dereceleri, Neighbour Joining (NJ) soy ağacıkullanılarak belirlenmeye çalışıldı. Analiz sonuçlarına göre; Agabus faldermanni (Zaitzev, 1927) sisteme kayıtlınükleotid dizileriyle %100 örtüşürken, bu değer; Hydroporus planus (Fabricius, 1782)’ta %99.88, Agabusbiguttatus (Olivier, 1795) ve Laccophilus minutus (Linnaeus, 1758)’ta %99.86, Agabus glacialis Hochhuth,1846’da %99.74, Laccophilus poecilus Klug, 1834’te %99.46, Agabus bipustulatus (Linnaeus, 1767)’ta %99.45,Agabus nebulosus (Forster, 1771)’ta %99.21, Hydroporus discretus Fairmaire & Brisout, 1859 ve Ilybiuschalconatus (Panzer, 1796)’ta %99.18, Agabus conspersus (Marsham, 1802)’ta %99.02, Bidessus calabricusGuignot, 1957’ta %98.94, Nebrioporus stearinus suavis (Sharp, 1882)’te %98.79, Hydroporus tessellatus(Drapiez, 1819)’ta %97.45, Liopterus haemorrhoidalis (Fabricius, 1787)’te %97.23, Hydroglyphus geminus(Fabricius, 1792)’te %96.47 ve Hydroporus palustris (Linnaeus, 1761)’te %93.54 oranlarında benzerliklerbelirlenmiştir.
Biosorption of 2,4-d, 2,4-DP, and 2,4-DB from aqueous solution by using thermophilic anoxybacillus flavithermus and analysis by high-performance thin layer chromatography: Equilibrium and kinetic studies
(Wiley, 2012) Özdemir, Sadin; Bekler, Fatma Matpan; Okumuş, Veysi; Dündar, Abdurrahman; Kılınç, Ersin; 0000-0001-8253-9568; 0000-0001-7384-7358; 0000-0002-5505-2700; 0000-0002-7930-1054; 0000-0001-5223-9919
In this study, the potential biosorption characteristics of the thermophilic Anoxybacillus flavithermus (A. flavithermus) was investigated for the removal of the chlorophenoxy acid derivates, namely, 2,4-dichlorophenoxy acetic acid (2,4-D), 2,4-dichlorophenoxy propanoic acid (2,4-DP or dichlorprop), and 2,4-dichlorophenoxy butyric acid (2,4-DB). The experiments were performed for the simultaneous biosorption of the studied pesticides. Optimum biosorption conditions were determined as a function of contact time, pH of the solution, amount of biomass, and initial pesticides concentrations. The concentrations of the pesticides in the remaining solutions were simultaneously analyzed by high performance thin layer chromatography. The optimum parameters were found as pH: 4.0 for biosorption medium, 60 min of contact time, 50 mg of bacteria, and 50 mg L-1 of initial pesticides concentrations. Langmuir and Freundlich models were applied to describe the biosorption isotherm of the pesticides by A. flavithermus as biomass. Biosorption of pesticides on to A. flavithermus showed pseudo first-order rate kinetics at different initial concentration of pesticides and different temperatures. The experimental adsorption data were fitted both the Langmuir and Freundlich adsorption models. Fourier-transform Infrared spectroscopy was used to understand the bonding mechanism of pesticides to biosorbent and surface functionality of the biosorbent The highest pesticide uptake was calculated from Langmuir isotherm and found to be 24.15 mg g-1 for 2,4-D. Among the studied pesticides, 2,4-DP showed difference adsorption behavior. According to in your comments the reason of this that 2,4-DP contain an asymmetric carbon atom, which provide a molecular chirality. (C) 2011 American Institute of Chemical Engineers Environ Prog, 2011
Characterisation of a partially purified protease from Bacillus cereus KG5 isolated from a hot spring
(Elsevier Sci Ltd, 2011) Ahmetoglu, Nazenin; Bekler, Fatma Matpan; Guven, Reyhan Gul; Acer, Omer; Guven, Kemal
[Abstract Not Available]
Characterization of a thermally stable ?-galactosidase produced by thermophilic anoxybacillus sp. AH1
(Bingöl Üniversitesi Fen Bilimleri Enstitüsü, 2021) Acer, Ömer; Bekler, Fatma Matpan
Thermostable β-galactosidases from thermophilic bacteria have attracted increasing interest to have various advantages in industrial and biotechnological applications. In this study, a highly thermally stable β-galactosidase produced by Anoxybacillus sp. AH1 was purified and characterized. The highest enzyme production was achieved after the bacterium was incubated for 24 hours. The enzyme was purified by precipitation with ammonium sulphate dialysis, gel filtration chromatography using Sephadex G-75. After the purification steps, β-galactosidase was found to be purified 10.2-fold and a yield of 13.9%. The molecular mass of the β-galactosidase was estimated to be 75 kDa by SDS-PAGE. The purified enzyme was highly stable and retained at 71% of the original activity at 60 °C and 53% at 70 oC within 120 minutes. The Km and Vmax values of purified β-galactosidase were calculated as 1.249 mM and 0.5 μmol minutes-1 , respectively. Ca2+, Zn2+, and Mg2+ significantly activated β-galactosidase activity, whereas enzyme activity was inhibited significantly by Cu+2 as well as by the metal ion chelators1,10- phenanthroline (phen) and ethylenediaminetetraacetic acid (EDTA). The Purified β-galactosidase activity was increased by PMSF (phenylmethylsulfonyl fluoride), PCMB (pchloromercuribenzoic acid), DTT (dithiothreitol), and β-ME (β-mercaptoethanol) at 2 mM, but inhibited completely by NEM (N-ethylmaleimide) at 1 mM.
Characterization of intracellular β-galactosidase from Bacillus subtilis 4NK and Bacillus paralicheniformis 5NK isolated from a hot water spring and effects of various inhibitors on enzyme activity
(Field Crops Central Research Institute, 2021) Tunç, Şaban; Bekler, Fatma Matpan; Güven, Kemal
In this study, the intracellular ?-galactosidases of Bacillus subtilis 4NK and Bacillus paralicheniformis 5NK isolated from Bingöl Binkap hot spring was partially purified and characterized. As a result of purification, the yield of the enzyme for B. subtilis 4NK was 85.2% and the purification fold was 2.8. The yield for B. paralicheniformis 5NK was 76.8% and the purification fold was 2.0. The optimum temperature of the enzyme was determined as 45oC for B. subtilis 4NK and 55oC for B. paralicheniformis 5NK and the optimum pH was 6.0 for both. In addition, in the thermal stability experiments even at the end of 120 min both enzymes were stable at 50oC. It was determined that the partially purified enzyme activity increased in the presence of iodoacetamide and phenylmethylsulfonylfluoride for B. subtilis 4NK, dithiothreitol, N-ethylenemaleimide and phenylmethylsulfonylfluoride for B. paralicheniformis 5NK. The metals were found to activate the enzyme at low concentrations of Co2+, Cd2+ and Mn2+ for B. subtilis 4NK, Cu2+ and Cd2+ were found to inhibit the enzyme at high rates for B. paralicheniformis 5NK. Km and Vmax values for 4NK and 5NK, respectively; 23.80 mM, 1.978 µmol/min and 5.61 mM, 1.869 µmol/min.
Cloning, purification and characterization of a thermostable β- galactosidase from Bacillus licheniformis strain KG9
(Cellular and Molecular Biology Association, 2015) Bekler, Fatma Matpan; Stougaard, Peter; Güven, Kemal; Güven, Reyhan Gül; Acer, Ömer
A thermo- and alkalitolerant Bacillus licheniformis KG9 isolated from Taslidere hot water spring in Batman/Turkey was found to produce a thermostable ?-galactosidase. Phylogenetic analysis showed that the 16S rRNA gene from B. licheniformis strain KG9 was 99.9% identical to that of the genome sequenced B. licheniformis strain DSM 13. Analysis of the B. licheniformis DSM 13 genomic sequence revealed four putative ?-galactosidase genes. PCR primers based on the genome sequence of strain DSM 13 were used to isolate the corresponding ?-galactosidase genes from B. licheniformis strain KG9. The calculated molecular weights of the ?-galactosidases I, II, III, and IV using sequencing data were 30, 79, 74, and 79 kDa, respectively. The genes were inserted into an expression vector and recombinant ?-galactosidase was produced in Escherichia coli. Of the four ?-galactosidase genes identified in strain KG9, three of them were expressed as active, intracellular enzymes in E. coli. One of the recombinant enzymes, ?-galactosidase III, was purified and characterized. Optimal temperature and pH was determined to be at 60 °C and pH 6.0, respectively. Km was determined to be 1.3 mM and 13.3 mM with oNPG (ortho-nitrophenyl-?-D-galactopyranoside) and lactose as substrates, respectively, and Vmax was measured to 1.96 ?mol/min and 1.55 ?mol/min with oNPG and lactose, respectively.
The Effects of Silver Nanoparticles (AgNPs) on Thermophilic Bacteria: Antibacterial, Morphological, Physiological and Biochemical Investigations
(Mdpi, 2024) Jahan, Israt; Bekler, Fatma Matpan; Tunc, Ahmed; Guven, Kemal
Since thermophilic microorganisms are valuable sources of thermostable enzymes, it is essential to recognize the potential toxicity of silver nanoparticles used in diverse industrial sectors. Thermophilic bacteria Geobacillus vulcani 2Cx, Bacillus licheniformis 3CA, Paenibacillus macerans 3CA1, Anoxybacillus ayderensis FMB1, and Bacillus paralicheniformis FMB2-1 were selected, and their MIC and MBC values were assessed by treatment with AgNPs in a range of 62.5-1500 mu g mL(-1). The growth inhibition curves showed that the G. vulcani 2Cx, and B. paralicheniformis FMB2-1 strains were more sensitive to AgNPs, demonstrating a reduction in population by 71.1% and 31.7% at 62.5 mu g mL(-1) and by 82.9% and 72.8% at 250 mu g mL(-1), respectively. TEM and FT-IR analysis revealed that AgNPs caused structural damage, cytoplasmic leakage, and disruption of cellular integrity. Furthermore, cell viability showed a significant decrease alongside an increase in superoxide radical (SOR; O-2(-)) production. beta-galactosidase biosynthesis decreased to 28.8% level at 500 mu g mL(-1) AgNPs for G. vulcani 2Cx, 32.2% at 250 mu g mL(-1) for A. ayderensis FMB1, and 38.8% only at 62.5 mu g mL(-1), but it was completely inhibited at 500 mu g mL(-1) for B. licheniformis 3CA. Moreover, B. paralicheniformis FMB2-1 showed a significant decrease to 11.2% at 125 mu g mL(-1). This study is the first to reveal the toxic effects of AgNPs on thermophilic bacteria.
Essential oil compositions and antimicrobial activities of Thymbra spicata L. var. spicata L., Lavandula X Intermedia Emeric ex Loisel., Satureja macrantha C. A. MEYER and Rosmarinus officinalis L.
(Inst Tecnologia Parana, 2022) Karakaş, Özgür; Bekler, Fatma Matpan
Medicinal and aromatic plants have been widely using in folk medicine as antimicrobial, anti-inflammatory and antinociceptive agents. The aim of this study was to determine essential oil composition and antimicrobial activity of T. spicata, L. X Intermedia, S. macrantha and R. officinalis. Essential oil components of these plants were obtained by water vapor distillation method using Neo-Clevenger apparatus. Essential oil components were determined by gas chromatography-mass spectroscopy (GC-MS). The main components of these plants are carvacrol (74.26 %) and gamma-terpinene (10.28%) in T. spicata, 1,8-cineol (32.48%), linalool (24.38%) and camphor (14.73%) in L. X Intermedia, p-cymene (56.70%), carvacrol (10.96 %) in S. macrantha and camphor (18.26 %), alpha-pinene (15.51%), 1,8-cineole (11.86%) and borneol (10.39%) in R. officinalis were determined. T. spicata and S. macrantha showed strong effects against three microorganisms. L. X lntermedia and R. officinalis showed strong activity against Candida albicans, while they had moderate effects against Staphylococcus aureus, Escherichia coli.
A highly inducible ?-galactosidase from enterobacter sp
(Serbian Chemical Society, 2020) Shaikhan, Bestoon Ahmed; Güven, Kemal; Bekler, Fatma Matpan; Acer, Ömer; Güven, Reyhan Gül
Enterobacter sp. 3TP2A isolated from a petroleum station was found to produce a novel, highly inducible mesophilic intracellular β-galactosidase in the presence of lactose up to 76.5 U mg-1. The enzyme was purified to 17.3-fold after gel permeation chromatography with a yield of approximately 11 %. The optimum pH and temperature values of the purified enzyme were found to be 8.0-9.0 and 35 °C, respectively. The molecular weight of the enzyme was approx. 60 kDa with a single band by both SDS-PAGE and native-PAGE, and estimated by gel filtration chromatography. The enzyme was inhibited by Zn2+ and EDTA, while Cu2+ had strong inhibitory effect even at low concentrations. Activation by Mg2+ and inhibition by EDTA show that the enzyme is metal-dependent or a metalloenzyme. The enzyme was slightly activated by 2-mercaptoethanol, while slightly inhibited by iodoacetamide. On the other hand, PCMB inhibited the enzymatic activity to a great extent, whereas it was completely inhibited by N-ethylmaleimide. The Vmax and Km values were calculated as 0.701 μmol min-1 and 0.104 mM, respectively. The results indicated that the β-galactosidase Enterobacter sp. 3TP2A might well be a good candidate for use in biotechnology, particularly in the area of environment and health.
Isolation and characterization of long-chain alkane-degrading Acinetobacter sp BT1A from oil-contaminated soil in Diyarbakir, in the Southeast of Turkey
(Taylor & Francis Inc, 2016) Acer, Omer; Guven, Kemal; Bekler, Fatma Matpan; Gul-Guven, Reyhan
A strain of long-chain alkane-degrading bacteria, BT1A, was isolated from oil-contaminated soil in Diyarbakr, in the southeast of Turkey. Morphological, biochemical, and physiological characterization and 16S rRNA gene sequence analysis showed that the strain BT1A was a member of Acinetobacter genus, and it was found to be closely related to Acinetobacter baumannii. The strain BT1A was able to utilize crude petroleum as carbon and energy sources in order to grow. Among the aliphatic hydrocarbons, growth was observed only in the medium containing long-chain alkanes (tridecane, pentadecane, and hexadecane) and squalene. Hexadecane was the most preferred hydrocarbon among the long-chain alkanes. Gas chromatography-mass spectrometry (GC-MS) analysis showed that BT1A degraded 83% of n-alkanes of 1% crude oil in 7days. The present study indicates that the isolated strain can well be used for biodegradation of hydrocarbons in oil-contaminated sites.
Isolation and characterization of long-chain alkane-degrading Acinetobacter sp. BT1A from oil-contaminated soil in Diyarbakir, in the Southeast of Turkey (vol 20, pg 80, 2016)
(Taylor & Francis Inc, 2017) Acer, Omer; Guven, Kemal; Bekler, Fatma Matpan; Gul-Guven, Reyhan
[Abstract Not Available]
Isolation and cloning of extracellular thermostable ?-galactosidases from a newly isolated Thermophilic Bacillus licheniformis KG9
(Elsevier Sci Ltd, 2011) Bekler, Fatma Matpan; Stougaard, Peter; Guven, Reyhan Gul; Guven, Kemal
[Abstract Not Available]
Isolation and identification of a novel thermo-alkalophilic Anoxybacillus sp. strain KB4 from Kuşburnu hot spring in Turkey
(2016) Bekler, Fatma Matpan
Termofiller 110 oC'ye kadar sıcaklıklarda ve farklı pH'larda (termo-asidofil ve termo-alkalofil) büyüyebilme yeteneğine sahiptirler ve biyoteknolojik uygulamaların ilgi çeken bir kaynağıdırlar. Bu çalışmanın amacı, Türkiye'deki Diyadin ilçesi Kuşburnu sıcak su kaynağından termofilik bakteri izole etmektir. KB4 suşu, morfolojik, fizyolojik, biyokimyasal testler ve 16S rRNA gen sekanslamayla tanımlanmıştır. KB4 suşu, aerobik, Gram-pozitif, çubuk şeklinde, hareketli ve sarı pigmentli olarak bulunmuştur. Optimum üreme pH 9.0-10.0'da, 55-60 oC'de ve %3 (w/v) NaCl konsantrasyonunda elde edilmiştir. Farklı karbon ve azot kaynaklarının kullanımı denendi. KB4 suşu, glukoz, galaktoz, sükroz, maltoz, arabinoz, ksiloz, yeast ekstrakt, pepton, tripton ve casamino asit gibi bazı karbon ve azot kaynaklarını kullanabilmektedir. 16S rRNA gen sekansına bağlı filogenetik analizler KB4 suşunun %98.78 sekans benzerliği ile Anoxybacillus pushchinoensis K1(T)'e oldukça yakın olduğu göstermektedir. Ek olarak, bu çalışma Türkiyede'ki jeothermal sulardan termofilik bakterilerin çeşitliliğini belirlemede rehber olacaktır.
Isolation and identification of petroleum degrading bacteria
(Elsevier Sci Ltd, 2011) Guven, Reyhan Gul; Bekler, Fatma Matpan; Kaya, Hasan; Acer, Omer; Guven, Kemal; Temel, Hamdi
[Abstract Not Available]
Isolation and production of thermostable ?-amylase from thermophilic Anoxybacillus sp KP1 from Diyadin hot spring in Agri, Turkey
(Springer, 2014) Bekler, Fatma Matpan; Guven, Kemal
A novel amylolytic enzyme producing thermophilic bacterial strain KP1 from the Diyadin hot spring water in Agri, Turkey, was isolated in the present study. Phylogenetic analysis based on the partial 16S rRNA gene, biochemical and physiological tests revealed that the strain KP1 belongs to the genus Anoxybacillus. The pH and temperature optima for the alpha-amylase production by Anoxybacillus sp. KP1 were 8.0 and 50A degrees C, respectively, where the maximum growth was obtained at the 28(th) hour of incubation and the highest alpha-amylase activity was obtained at the 40(th) hour of incubation (8979.6 U/mL). The optimum pH and temperature for the enzyme activity were 8.0 and 60A degrees C, respectively. The maximum alpha-amylase production was secreted in the presence of 2% (w/v) soluble starch (10837.7 U/mL). Among the various organic and inorganic nitrogen sources tested, while keeping the beef extract concentration constant, casamino acid (14310.6 U/mL), urea (14126 U/mL), and tryptone (13217.2 U/mL) at a concentration of 2% gave the maximum alpha-amylase production. The enzyme activity was enhanced in the presence of 1.5 mM Mn2+ (123%), whereas it was strongly inhibited 1.5 mM by Hg2+. Inhibition by 89% was obtained also with sodium dodecyl sulphate (1%). The enzyme was found to be relatively stable at a range of pH and temperature.
Molecular characterisation and numerical analysis of novel moderately thermophile Anoxybacillus sp. FMB1
(Ars Docendi, 2018) Bekler, Fatma Matpan; Yalaz, Secil; Guven, Kemal
A moderately thermophilic bacteria was isolated from the water sample collected from Yozgat, in the Central Anatolia Region of Turkey. Morphological, physiological and biochemical characteristics of thermophilic isolate were determined in comparison to standard type strains, and they were all identified and classified into homogenous groups by using fundamental principles of numerical taxonomy. The UPGMA method was used for numerical taxonomy and the phenogram was drawn with MATLAB. Phylogenetic analyses were also performed by the neighbour-joining and UPGMA methods. According to the numerical definition result and 16S rRNA analysis, the strain FMB1 was most closely related to Anoxybacillus ayderensis AB04(T). The results showed that MATLAB computer program using UPGMA method can be used as an alternative or additional method in bacterial classification, before 16S rRNA analysis performed.
Optimization of the thermostable alkaline and Ca-dependent ?-amylase production from Bacillus paralicheniformis by statistical modeling
(Serbian Chemical Society, 2019) Bekler, Fatma Matpan; Yalaz, Seçi̇l; Güven, Reyhan Gül; Güven, Kemal
A novel amylolytic enzyme producing thermoalkaliphilic bacterium, the source of industrially used enzymes was isolated. Isolated strain was identified by morphological, physio-biochemical tests and the 16S rRNA gene sequence analysis. The optimal conditions of enzyme activity were determined. For higher α-amylase production, the variables such as yeast extract, starch, CaCl2, (NH4)2SO4, NaCl and MgSO4 in the α-amylase production medium, the temperature and pH were screened by Plackett–Burman design and optimised using response surface methodology (RSM). The optimal conditions were found to be 0.15 g/L for starch, 0.15 mg/L for CaCl2 and 60 °C for temperature. By using RSM model, amylase production increase was achieved sevenfold. It is showed that this method can be utilised to optimize α-amylase production in athermophilic bacteria such as Bacillus paralicheniformis. Keywords: α-amylase; Bacillus paralicheniformis; optimization; response surface methodology.
PCR-based detection of alkane monooxygenase genes in the hydrocarbon and crude oil-degrading Acinetobacter strains from petroleum-contaminated soils
(Serbian Chemical Society, 2024) Eren, Ayşe; Bekler, Fatma Matpan; Güven, Kemal
Bacterial strains D11, E1 and E2 isolated from petroleum-contaminated soils were found to be members of Acinetobacter genus revealed by 16S rRNA gene sequence analysis and phenotypic characteristics. After incubation for 5 days, about 43, 9 and 12 % of total petroleum hydrocarbons of crude oil were degraded by strains D11, E1 and E2, respectively, and determined by GC-MS analysis. Moreover, about 70 and 76 % of single hydrocarbon hexadecane was degraded by the strains D11 and E1 after 3 days of short incubation time, respectively, while the strain E2 degraded about 48 % of single hydrocarbon pentadecane. By using PCR-based method, gene sequences of the strains D11 and E2 showed similarity to alkane 1-monooxygenases from Acinetobacter sp. BUU8 alkM with 93.06 and 92.72 %, respectively, while the sequence similarity of strain E1 was 95.84 % to Acinetobacter sp. 826659. The present study of hydrocarbon biodegradation by Acinetobacter strains may provide a good advantage in bioremediation process. © 2024 Physical Society of Japan. All rights reserved.

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