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Öğe Alpha-Amylase Production by Bacillus subtilis RSKK96 in Submerged Cultivation(Kafkas Univ, Veteriner Fakultesi Dergisi, 2011) Akcan, Nurullah; Uyar, Fikret; Guven, AyselThe bacterial strain Bacillus subtilis RSKK96 was shown to produce extracellular alpha-amylase. Enzyme synthesis occurred at 72 h with an optimum of 37 degrees C. Effects of various carbon, nitrogen, amino acid sources and chemicals on alpha-amylase production were examined. FeSO4, ZnSO4 and CuSO4 inhibited bacterial growth as a result, amylase production. Maximum alpha-amylase production (858.6 +/- 41.9 U/mg) was obtained in a medium containing 0.5% cotton stalk in 72 h.Öğe Antimicrobial activities of some thyme (Thymus, Staureja, Origanum and Thymbra) species against important plant pathogens (vol 18, pg 1455, 2006)(Mrs Pushpa Agarwal, 2006) Kizil, Suleyman; Uyar, Fikret[Abstract Not Available]Öğe Bacillus subtilis'te üreme, plazmid ve α-amilaz sekresyonu üzerine kalsiyum kanal blokörlerinin ve ağır metallerin etkisi(2018) Uyar, Fikret; Ensari, N. YavuzBu çalışmada Nottingham Üniversitesi Tıp Fakültesi Mikrobiyoloji Bölümü'nden ve Ankara Üniversitesi Ziraat Fakültesi Gıda Mühendisliği Bölümü'nden temin edilen B. subtilis kullanıldı. Bakterilerin Nutrient Broth (NB) ve Starch Beef (SB) ortamında daha iyi üreme gösterdiği saptanarak, bu besi ortamlarında a-amilaz sekresyonunun hangi sürede maksimuma ulaştığı aktivite tayin yöntemi ile tespit edilip elektroforez ile teyit edildi. Bakteri, SB sıvı besi yerinde çeşitli kalsiyum kanal blokörleri ile kontrol eşliğinde etkileştirilerek, bu blokörlerin, üreme, a-amilaz üretimi ve bakteri plazmitleri üzerine etkileri araştırıldı. Blokörlerin üreme ve a-amilaz sekresyonunu inhibe ettiği saptandı. Kalsiyum kanal blokörlerinin a-amilaz enzimi üzerine in vitro etkisinin olmadığı gözlendi. Bakteri, SB sıvı besi yerinde ağır metaller ile beraber kalsiyum kanal blokörü olan Diltiazem ile kontrol eşliğinde etkileştirilerek, üreme, a-amilaz üretimi ve bakteri plazmitleri üzerine etkileri araştırıldı. HgCk'nin üreme ve a-amilaz sekresyonunu inhibe ettiği saptandı. ZnC^'nin üreme ve amilaz sekresyonu üzerinde etkili olmadığı, Diltiazem' in yalmz başına plazmit üzerine bir etkisinin olmadığı, ancak her iki ağır metalin Diltiazem eşliğinde plazmit amplifikasyonuna neden olduğu saptandı. NB sıvı besi yerinde bakteri üretiminin belirlenen zaman diliminde ortama ilave edilen Chloramphenicol ile protein sentezi durdurularak, membrandaki ve salgılanmış enzim aktivitelerine göre a-amilazm sentez ve olgun hale geliş süreleri belirlendi. İnhibisyonun tamamlanıp ribozom üzerinden membrandan çıkış süresi 70-80 dakika olarak tespit edildi. pH etkisine maruz bırakılan nötral proteaz enziminin, aktivitesinde meydana gelen değişiklikler belirlendi. Enzimin maksimum aktiviteyi pH 7'de gösterdiği ve ı/y dializ ile prostetik grubu olan Zn 'nin enzimden ayrılmadığı saptandı. Optimal dışındaki pH'larda enzimin üç boyutlu yapısının pH'ya bağımlı bozulması ile aktivitesini kaybettiği saptandı.Öğe The effects of amino acids on production and transport of α-amylase through bacterial membranes(Wiley-VCH Verlag, 2000) Agüloǧlu, Sema; Ensari, N. Yavuz; Uyar, Fikret; Otludil, Birol; 0000-0002-3809-5987Bacillus subtilis was grown in media containing various amino acids. The effects of these aminoacids on the production of secreted alpha-amylase and membrane-bound alpha-amylase were investigated. Their effects on the transport of alpha-amylase through membranes were also determined. The synthesis rate of alpha-amylase isolated from bacterial membranes was stimulated by growth in media separately containing Ala and Arg, all other amino acids being inhibitory. Cys, Arg and Leu increased the transport of alpha-amylase through the membrane, while Ala, Asp and Ser inhibited it. Secreted alpha-amylase activity was appreciably stimulated by Ala, Arg, Gin, Gly, Leu, Phe, Pro, and especially Cys while it was repressed by Asp, Asn, Glu, Lys, Met, Ser, Thr and Trp.Öğe The effects of mucosal media on some pathogenic traits of Crohn's disease-associated Escherichia coli LF82(Future Medicine Ltd, 2018) Aygun, Husamettin; Karamese, Murat; Ozic, Cem; Uyar, FikretAim: Adherent-invasive Escherichia coli (AIEC) pathovar has been identified in intestinal mucosa of patients with Crohn's disease. Our aim was to compare the impact of sterile mucosal media (Muc-M) originated from different parts of the intestine on some pathogenic traits of AIEC LF82 strain. Materials & methods: Muc-M composed of certain rates of cell culture medium or M63 minimal medium and mucosal contents obtained from different part of intestine were designed for cell-infection experiments and biofilm-formation assays. Results: The results showed that Muc-M reduced usually pathogenic properties of AIEC LF82. However, LF82 adhesion, invasion and specific biofilm formations were markedly higher in Muc-MCR than those in Muc-MIR. Conclusion: In this context, the findings of present study could help the endeavors related to determining molecular targets for AIEC bacteria.Öğe The Inhibitory Effect of Ileal Mucosal Media Originated from FVB/N mice strain on Escherichia coli LF82 Invasion(Bezmialem Vakif Univ, 2019) Aygun, Husamettin; Karamese, Murat; Uyar, FikretObjective: The aim of this study was to investigate the effect of healthy mucosa on adhesive and invasive properties of AIEC reference strain Escherichia cell (E. coli) LF82. For this purpose, we had designed special medias that contained cell culture medium and mucosal content obtained from different regions (colon and ileum) of the digestive tract. Methods: We tested the infecting ability of AIEC reference strain E. coli LF82 on 1-407 cells in the presence of mucosal media (Muc-M) under in ritro conditions. Muc-M composed of certain rates of cell culture medium or M63 minimal medium and mucosal contents obtained from different part of intestine were designed for cell-infection experiments and biofilm-formation assays. Results: The result showed that the mucosal media decreased the infection percentage of E. coli LF82 strain when compared with control group. It was seen that the mucosal media originating from ileum almost completely inhibited the invasion of LF82 strain. On the other hand, it was observed that the mucosal media prepared from colon) reduced the bacterial invasion only in half the rate when compared with control. Conclusion: The findings showed that these medias obtained from different regions of the intestinal tract affected LF82 invasion at different rates. Therefore, this study provided crucial information that could contribute to the future studies on the localization of bacteriaÖğe Optimal conditions for production of extracellular protease from newly isolated Bacillus cereus strain CA15(2011) Yılmaz, Ebru İnce; Porsuk, Ilknur; Uyar, Fikret; Kızıl, GökselTopraktan alkalen proteaz üreticisi Bacillus sp. CA15 izole edildi. 16S ribosomal RNA gen dizilimine gore bu mikroorganizmanin Bacillus cereus ile çok yakin akraba oldugu tespit edildi. Proteaz üretiminin artisini saglamak için, kültür kosulari; karbon ve azot kaynaklari, metal iyonlari, pH ve isinin etkisi açisindan optimize edildi. En yüksek proteaz üretimi, %1 yagsiz süt tozu, %1 çözünür nisasta ve %0.6 $MgSO _4.7H _2O$ varliginda, baslangiç pH'si 8.0 ve 35°C'de üretilen kültürlerden elde edildi. Bu besiyerinde üretilen hücrelerin, canli hücre sayisinin, mL'de 1.8x108'e vardigi duragan fazda en iyi enzim aktivitesine rastlandi. Inorganik azot kaynaklari varliginda proteaz seviyesinin oldukça düsük oldugu bulundu. Sükroz ve laktozun varliginda da proteaz üretiminin azaldigi tespit edildi. Bacillus sp. CA15 alkalen proteazinin Triton X-100, Tween 20 and SDS'ye karsi son derece dayanikli oldugu gözlemlendi. Enzim aktivitesinin PMSF varliginda inhibe olmasi bu enzimin bir serin proteaz oldugunu düsündürdü.Öğe Optimization and enhanced production of ?-amylase and protease by a newly isolated Bacillus licheniformis ZB-05 under solid-state fermentation(Springer, 2013) Karatas, Hakan; Uyar, Fikret; Tolan, Veysel; Baysal, ZubeydeEight different agro-residues were tested for alpha-amylase and protease production by using Bacillus licheniformis ZB-05. Among them, rice husk (RH) was proved as the best substrate for two enzymes (alpha-amylase 443 U/g and protease 469,000 U/g). Maximum enzyme production was observed to be 30 % initial moisture, with a growth period of 36 h in 20 and 30 % inoculum volumes for alpha-amylase and protease, respectively. The best enzyme recovery from solid mass was obtained when extracted with tap water. Among the tested various nitrogen sources, 1 % ammonium sulphate followed by 2 % Bacto liver, 2 % ammonium sulphate and 1 % Bacto casaminoacid served as the best inorganic and organic nitrogen sources for alpha-amylase and protease production, respectively. As additional carbon sources, 2 % soluble starch enhanced alpha-amylase production, while 1 % maltose enhanced protease production.Öğe OPTIMIZATION OF ?-AMYLASE PRODUCTION BY Bacillus subtilis RSKK96: USING THE TAGUCHI EXPERIMENTAL DESIGN APPROACH(Taylor & Francis Inc, 2011) Uysal, Ersin; Akcan, Nurullah; Baysal, Zubeyde; Uyar, FikretIn this study, the Taguchi experimental design was applied to optimize the conditions for alpha-amylase production by Bacillus subtilis RSKK96, which was purchased from Refik Saydam Hifzissihha Industry (RSHM). Four factors, namely, carbon source, nitrogen source, amino acid, and fermentation time, each at four levels, were selected, and an orthogonal array layout of L-16(4(5)) was performed. The model equation obtained was validated experimentally at maximum casein (1%), corn meal (1%), and glutamic acid (0.01%) concentrations with incubation time to 72 h in the presence of 1% inoculum density. Point prediction of the design showed that maximum alpha-amylase production of 503.26 U/ mg was achieved under optimal experimental conditions.Öğe Production and characterization of neutral and alkaline protease from different Bacillus subtilis strains(Asian Journal Of Chemistry, 2008) Uyar, Fikret; Baysal, ZuebeydeAn extracellular neutral and alkaline protease from two different Bacillus subtilis strains were studied. The optimal activity occured when the pH level was 7.0 and 10.5 at a temperature of 35 and 45 degrees C for neutral protease and alkaline protease, respectively. When neutral protease was stable in the temperature range 35-60 degrees C, alkaline protease was found stable between 35-55 degrees C for 0.5 h. Divalent cations, especially Ca2+ increased enzymes activity and were inhibited by Mn2+,Ni2+, CU2+, Fe2+, Co2+, Cd2+ and Hg2+ for neutral protease and by Zn2+, Cd2+,Co2+, Cu2+ and Hg2+ for alkaline protease. The neutral protease was also inhibited by ethylenediamine-tetraacetic acid, 1, 10 phenanthroline and dithiothreitol whereas alkaline protease was inhibited with phenylmethyl-sulfonyl fluoride. The obtained K-m values were 2.28 x 10(-3) +/- 3.65 x 10(-4), 2.28 x 10(-4) +/- 4.21 x 10(-5) and 1.70 x 10(-4) +/- 5.18 x 10(-5) M for azocasein, BSA and casein for neutral protease, respectively. The K-m values with azocasein, BSA, casein, N-suc-Ala-Ala-Ala-pNA and N-cbz-Ala-Ala-Leu-pNA were 2.02 x 10(-3) +/- 7.3 x 10(-5), 2.13 x 10(-4) +/- 6.04 x 10(-5), 8.75 x 10(-4) +/- 1.36 x 10(-4), 1.71 x 10(-3) +/- 3.93 x 10(-4) and 2.64 x 10(-4) +/- 4.93 x 10(-5) for alkaline protease.Öğe Production and optimization of ?-amylase from Bacillus circulans ATCC 4516 with solid state fermentation(2012) Uyar, Fikret; Akcan, Nurullah; Serin, BesiBu çalışmanın amacı Bacillus circulans ATCC 4516 tarafından üretilen ?-amilazın belirli üretim parametrelerini incelemektir. ?-Amilazın optimizasyon parametreleri katı faz fermantasyonu ile gerçekleştirilmiştir. Katı atık besin kaynağı olarak pirinç kepeği kullanılmıştır. Karbon ve metal tuz kaynakları eklenmesi ile enzim üretimi azalmıştır. İnkübasyon zamanı, inkübasyon sıcaklığı, inokülüm düzeyi, başlangıç pH ve ekstraksiyon ortamı gibi belirli fermantasyon parametreleri ayrı ayrı incelenmiştir. Maksimum ?-amilaz üretim miktarı (2716.9 ± 35.9 U/mg) amonyum klorür eklenmesi ile 48. saatte 37ºC’de inokülüm düzeyi %25, başlangıç pH’ı 7.5 olarak elde edilmiştir.Öğe Production and Optimization of ?-Amylase from Bacillus circulans ATCC 4516 with Solid State Fermentation ?(Hacettepe Üniversitesi, 2012) Serin, Besi; Akcan, Nurullah; Uyar, FikretThe aim of this paper is to study influence of the certain production parameters of ?-amylase by Bacillus circulans ATCC 4516. Optimization parameters of ?-amylase were carried out with solid state fermentation SSF . Solid waste from rice bran used as the basic nutrient source. Supplementation with carbon and metal salt sources decrased the enzyme production. Certain fermentation parameters involving incubation time, incubation temperature, inoculum level, initial pH and extraction medium were studied separately. Maximal amount of ?-amylase production 2716.9 ± 35.9 U/mg was obtained inoculum level 25%, initial pH 7.5 at 37oC for 48 h with supplementation of ammonium chloride.Öğe Production of extracellular alkaline ?-amylase by solid state fermentation with a newly isolated Bacillus sp.(Taylor & Francis Inc, 2008) Baysal, Zuebeyde; Uyar, Fikret; Dogru, Mehmet; Alkan, HueseyinProduction of alkaline -amylase employing our laboratory isolate, Bacillus sp., under solid state fermentation, was optimized. The effect of wheat bran and lentil husk was examined. Lentil husk exhibited the highest enzyme production. The appropriate incubation time, inoculum size, moisture level, and buffer solution level were determined. Maximum yields of 216,000 and 172,800 U/g were achieved by employing lentil husk and wheat bran as substrates in 0.1 M carbonate/bicarbonate buffer at pH 10.0 with 30% initial moisture level at 24h. Inoculum size and buffer solution level were found to be 20% and 1:0.5 for two solid substrates.Öğe Production of extracellular alkaline protease from Bacillus subtilis RSKK96 with solid state fermentation(2011) Uyar, Fikret; Akcan, NurullahBu çalismada, kati faz fermantasyonu ile Bacillus subtilis RSKK96'dan ekstrasellüler alkalin proteaz üretimi incelenmistir. Enzim üretimi için farkli tarim atiklari substrat olarak kullanilmistir. Tarim atiklari arasinda mercimek kabugu proteaz üretimini (3937.0 U/mg) desteklemistir. Üretim parametleri inkübasyon zamani 120 saat, ekstraksiyon medyumu %1 triton-X100, baslangiç nem içerigi %30, baslangiç pH 9.0 olarak optimize edilmistir. Arabinoz, içeren ortamda yüksek alkalin proteaz üretimi gerçeklesmistir, bunu laktoz, galaktoz ve fruktoz izlemistir. Farkli azot kaynaklari arasinda et özütü alkalin protez üretimini en iyi sekilde indüklemis iken diger azot kaynaklari enzim üretimini baskilamistir. Metal tuzlari arasinda $FeSO _4.7H _2O$ ve $MgSO _4.7H _2O$ proteaz üretimini arttirmistir. En yüksek enzim üretimi (5759.2 U/mg) 1000 mL fermantasyon ortaminda gerçeklesmistir.Öğe Production of lipase by a newly isolated Bacillus coagulans under solid-state fermentation using melon wastes(Humana Press Inc, 2007) Alkan, Huseyin; Baysal, Zubeyde; Uyar, Fikret; Dogru, MehmetAn extracellular lipase was produced by Bacillus coagulans by solid-state fermentation. Solid waste from melon was used as the basic nutrient source and was supplemented with olive oil. The highest lipase production (78,069 U/g) was achieved after 24 h of cultivation with 1% olive oil enrichment. Enzyme had an optimal activity at 37 degrees C and pH 7.0, and sodium dodecyl sulfate increased lipase activity. NH4NO3 increased enzyme production, whereas organic nitrogen had no effect. The effect of the type of carbon sources on lipolytic enzyme production was also studied. The best results were obtained with starch and maltose (148,932 and 141,629 U/g, respectively), whereas a rather low enzyme activity was found in cultures grown on glucose and galactose (approx 118,769 and 123,622 U/g, respectively). Enzyme was inhibited with Mn+2 and Ni+2 by 68 and 74%, respectively. By contrast, Ca+2 enhanced enzyme production by 5%.Öğe Prokaryotlarda proteolitik aktivite(2018) Uyar, Fikret; Ensari, YavuzBu çalışmada Çermik Termal kaplıcalarından izole edilerek tür teşhisi yapılan termotoleran Bacillus subtilis kullanıldı. B.subtilis'in hangi besi ortamında daha iyi Nötral proteaz ürettiği saptanarak bu ortamdan nötral proteazın saflaştırmasına gidildi. Yapılan deneylerde 10~5 M Zn2+ içeren nutrient broth besiyerinde üretilen B. subtilis'in salgıladığı nötral proteazın hem aktivite yönünden, hem de salgılanan enzim miktarı bakımından en uygun besi ortamı olduğu saptandı.Saf olarak elde edilen Nötral proteaz için molekül ağırlığı 29.000 Dalton, Km değeri de 0.952 olarak belirlendi.Öğe Screening of Various Organic Substrates and the Development of a Suitable Low-Cost Fermentation Medium for the Production of ?-Amylase by Bacillus subtilis(Faculty Food Technology Biotechnology, 2009) Ozdemir, Sadin; Guven, Kemal; Baysal, Zuebeyde; Uyar, FikretThe production of extracellular amylase by Bacillus subtilis has been studied in solid-state fermentation (SSF). In a sequential order, various process parameters were optimized for maximum amylase production. The tested process parameters were different solid substrates such as banana husk (BH), water melon husk (WMH), lentil bran (LB), wheat bran (WB), melon husk (MH) and maize oil cake (MOC), different incubation time (24-144 h), particle size (500-2500 mu m), initial moisture content of the substrate (40-70 %, by mass per Volume), inoclum size (10-60 %, by mass per volume) and inoculum concentration (10-60 % by volume per mass). The maximum production of amylase (4857 U/mg) was achieved when 1 % starch (by mass, particle size of 1500 pm) was added to banana husk as the solid substrate, fermented for 72 h at 37 degrees C, at an inoculum level of 30 % (by volume per mass), and initial moisture content of 60 % (by volume per mass)Öğe Submerged Kültürü ile Bacillus subtilis RSKK96'dan ?-Amilaz Üretimi(2011) Uyar, Fikret; Güven, Aysel; Akcan, NurullahBakteriyel tür Bacillus subtilis RSKK96'nın ekstrasellüler ?-amilaz ürettiği gözlenmiştir. Enzim sentezi 72. saatte optimum 37°C'de elde edilmiştir. Çeşitli karbon, azot ve aminoasit kaynakları ve kimyasalların ?-amilaz üretimi üzerine etkileri incelenmiştir. FeSO4, ZnSO4ve CuSO4 bakteriyel büyümeyi ve dolayısıyla ?-amilaz üretmini inhibe etmiştir. Maksimum ?-amilaz üretimi (858.6±41.9 U/mg) %0.5 pamuk sapı bulunan ortamda 72. saatte elde edilmiştir.
