Hastane ve şehir atık sularından izole edilen Enterobacteriacea grubu bakterilerde II.kuşak cephalosporin dirençliliğinin devamı ve farklı sıcaklıklardaki hareketli ve durgun besi ortamları ile zoogloea ramıgeranın plasmid transferlerine etkileri

TEZ829Tez (Doktora) -- Çukurova Üniversitesi, Adana, 1990.Kaynakça (s. 99-109) var.iv, 109 s. ; rnk.res. 30 cm.

Flokulasyonun tekstil atıksularının arıtılmasına ve Escherichia Coli el iminasyonuna etkisi

TEZ379Tez (Yüksek Lisans) -- Çukurova Üniversitesi, Adana, 1987.Kaynakça (s.52-54) var.ii, 56 s. ; 30 cm.

Alkaline, Thermostable,Thermophilic and Chelator Resistant Type II Pullulanase from Native Isolate Anoxybacillus rupiensis strain NP2

Pullulanase producing Anoxybacillus rupiensis strain NP2 (NCBI accession number: MF355418) was isolated from soil and screened for pullulanase activity on agar medium containing pullulan (pH 5.0) at 55°C. The highest enzyme production was optimally occurred at 55°C and pH 5.0 for 48 h. The enzyme (from supernatant) exhibited its maximal activity at 60°C and pH 9.0. It was highly stable between pH 3.0-11.0 for 24 h, maximally at pH 6.0 (131%) and 30-100°C for 60 min, maximally at 80°C (115%). The enzyme was treated with two each concentrations of various metal ions, chelators, detergents and inhibitors. 5 mM concentration of Ca2+ had a very strong stimulating effect on enzyme (176%). It showed 240% activity with 2-mercaptoethanol (%1) and remained 78% of its activity by Zn2+(10 mM). In the presence of 5 mM and 10 mM EDTA, NA2 pullulanase exhibited 193% and 84% activity, respectively. At 1% SDS, 83% enzyme activity was measured. According to TLC analysis, the end products of pullulan and starch are glucose, maltose, maltotriose and maltodextrin. This shows that NP2 amylase is an type II pullulanase (amyllopullulanase.) In conclusion, type II pullulanase producing by Anoxybacillus rupiensis strain NP2 is an alkaline, a thermo-stable enzyme and resistant to chelators and detergents. Because of its thermo-alkaline activity, it can be used in toothpaste, detergent and starch industry

Isolation Of Moderately Halofilic Bacillus Sp. Producing Amilase, Cellulase And Xylanase And Determination Of Optimum Growth And Enzyme Production

Van gölü kıyı şeridinden 20 farklı bölgeden alınan toprak örneklerinde toplam 242 bakteri izolasyonu yapılmış ve bunlardan 76’sı amilaz üreten (%31.66), 35’i selülaz üreten (%14.6), 32’si ksilanaz üreten (%13.2) özellikte suş olduğu belirlenmiştir. Bu suşlar içinden en iyi üreme ve enzim sentezini gösteren suşlar seçilerek identifiye edilmiştir. Amilaz üretimi için seçilen katı besiyerinde Bacillus sp.AB-17 suşu en iyi pH 9.5’de 37oC’de ve %7 tuz konsantrasyonunda üreme ve enzim üretimini gerçekleştirdiği bulunmuştur. Selülaz için seçilen Bacillus sp.C-14 suşu, pH 8.5-9.5 aralığında 37oC’de %3 tuz konsantrasyonunda üreme ve enzim üretimini gerçekleştirdiği saptanmıştır. Ksilanaz enzimi için seçilen Bacillus sp.X-13 suşu ise pH 6.0-7.0 aralığında 40oC’de en iyi üreme ve enzim sentezlediği saptanmıştır

Partial Purification and Characterization of an Acidophilic and a Thermophilic Amylase from Native Isolate Bacillus sp.

α-Amylase (α-1,4-glucan-4-glucanohydrolases, EC3.2.1.1) is an endoacting enzyme that catalyzes the hydrolysis of α-1,4-glycosidic linkages in starch and other related polysaccharides. It is one of the most important industrial enzymes with widespread applications in starch, baking, textile, brewing and paper industries. In the starch industry natural pH of starch slurry is usually around 4.5. Starch saccharification is totally enzyme-based. Acid-stable extracellular enzymes from microorganisms are required because they are utilized in the degradation of polymeric or oligomeric carbon sources which occurs at a pH ranges from 3.2 to 4.5. The promising property of enzymes from thermoacidophiles is activity at low pH and high temperature. This research was aimed to produce, partially purificate and characterize a thermophilic and an acidic amylase by native Bacillus sp. and find out biotechnological and industrial areas of usage of this enzyme. Bacillus sp. strains were isolated from soil samples and screened for amylase production on M9-starch agar (pH 5.0) at 55°C. The best acidic amylase producing isolate was incubated at different temperatures (35-70°C) and pH values (3.0-9.0) to find optimum enzyme production conditions. The selected strain was incubated at 55°C for 48 h in M9-starch medium (pH 5.0). After centrifugation, the supernatant was subjected to partially enzyme purification by ethanol precipitation. The optimum pH and temprature of amylase was tested at several pH buffers (3.0-11.0) and different temperatures (30-100°C). For pH and thermal stabilities, the enzyme was pre-incubated at pH values between pH 3.0-11.0 for 24 h and at temperatures between 30-100°C for 60 min. A thermophilic and an acidic amylase was produced by native Bacillus sp. Enzyme synthesis was optimally occurred at 55°C and pH 5.0. The enzyme is highly active in a broad temperature range between 30-100ºC, with an optimum at 60ºC. It was active between pH 3.0-11.0 with an maximum activity at pH 4.0. The enzyme also presented the thermo and acido stable properties. Our results are in accordance with the other researchs. According to Liu and Xu (2008), α-amylase from a newly isolated Bacillus sp. YX-1 exhibited maximum activity at pH 5.0, performed stability over a broad range of pH 4.5–11.0 and was optimally active at 40–50°C. Emtanani et al. (2014) showed that an extracellular recombinant α-amylase from Bacillus subtilis DR8806 exhibited optimal activity at pH 5.0 and 70°C with high stability in pH and temperature ranges of 4.0–9.0 and 45–75°C. Liu et al. (2015) reported that B. subtilis ZJ-1 produced an acidic α-amylase with considerable thermostability and the enzyme reached its maximum activity at pH 5.0 and 50°C. In conclusion, amylase produced by native isolate Bacillus sp. is an acidophilic, a thermophilic and a stable enzyme. Because of its thermo-acidophilic activity at low pH and high temperature, this enzyme can be used in starch, textile, fruit juice industries, traditional brewing and food processing where temperatures of pasteurization could be used to inactivate the enzymes after fermentation. This research (project code: FDK-2014-2393) was supported by Cukurova University Scientific Research Projects Coordination Unit and TÜBİTAK-BİDEB 2211-C

Optimization of Tyrosinase Enzyme Production from Native Bacillus sp. MV29 Isolate

Tyrosinase is a copper-containing enzyme which catalyzes the conversion of L-tyrosine to L-DOPA and melanin. L-DOPA is a preferred drug for treatment of Parkinson's disease and melanin has many pharmaceutically and therapeutically uses. In this research a native Bacillus sp. having tyrosinase enzyme was isolated from soil sample and the best carbon and nitrogen source for the enzyme production was found by analyzing effect of various carbon and nitrogen sources on the yield, also the optimum amount of the sources was selected by testing various amount of them. Various amounts of L-tyrosine and trace element were studied and the optimum amount of them was opted. Enzyme production by the native bacillus sp. was analyzed at different pH and temperature and the optimum pH value and temperature for the enzyme production was selected. By testing the enzyme activity for 56 hour at 4 hours intervals, the optimum incubation time was determined as 72 hour. According to the findings of this research, 40ᴼC as optimum temperature, pH 7.0 as optimum pH, 48h as optimum incubation time was determined as optimum for the enzyme production. The addition of L-tyrosine (5mM) as a substrate to the production medium was highly effective on enzyme production. Consequently, after optimization, 0.7IU tyrosinase enzyme per milliliter of medium culture was obtained

TERMOALKALİFİLİK AMİLAZ VE SELÜLAZ ENZİM (MULTİENZİM) ÜRETİCİSİ BACILLUS sp. İZOLASYONU, ENZİMLERİN KARAKTERİZASYONU VE BİYOTEKNOLOJİK UYGULANABİLİRLİĞİ

Bu çalışmada, doğal ortamdan izole edilen Bacillus sp. SHK-7 suşundan termoalkalifilik amilaz ve selülaz (multienzim) enzimlerinin üretimi ve karakterizasyonu gerçekleştirilmiştir. Bu amaçla, optimum aktivite sıcaklığı, optimum pH, pH stabilitesi ve sıcaklık stabilitesi analizleri gerçekleştirilmiştir. Amilaz enzimi optimum aktivitesini pH 8.0 ve 80°C’de göstermiştir. Enzim 30-100ºC arasında 60 dakikada aktivitesinin yaklaşık %93’ünü korumuştur. Amilaz enzimi 50ºC’de pH 8.0-13.0 aralığında 24 saat süreyle ortalama %84 stabil kalmıştır. Selülaz enzimi optimum aktivitesini 60°C ve pH 8.0’de göstermiştir. Enzim 30-100°C arasında 60 dakikada aktivitesinin yaklaşık %98 ini korumuştur. Selülaz enzimi 50°C’de pH 9.0-12.0 aralığında 24 saat süreyle ortalama %72 stabil kalmıştır. Bu sonuçlara göre SHK-7 amilaz ve selülaz enzimleri termofil, alkalifil, termostabil ve alkali-stabil özellik göstermekte olup başta deterjan endüstrisi olmak üzere tekstil, gıda ve içecek endüstrilerinde kullanılabilir özelliktedir

Bacillus sp. X13 suşundan multifonksiyonel endoksilanaz üretimi ve karakterizasyonu

Topraktan multifonksiyonel endoksilanaz üreten Bacillus sp. X13 suşu izole edilmiştir. Enzim sentezi optimum 37 °C olmak üzere 20 ile 60 °C arasında gerçekleşmiştir. SDS-PAGE analizinde, 66,5 kDa, 80,6 kDa, 95,5 kDa ve 108,4 kDa olarak hesaplanan 4 aktif enzim bandı gözlenmiştir. Enzim optimum aktivitesi 40 °C olup 20-90 °C arasında geniş bir sıcaklık aralığına sahiptir ve maksimum aktivitesi ise pH 6,0 olarak bulunmuştur. İnkübasyon sıcaklığı arttıkça enzim kalan aktivitede giderek artan bir düşüş göstermiştir. Sıcaklık stabilitesi Ca+2 varlığında dahi artmamıştır. Enzim pH 3,6 ile 10,0 arasında 1 saat inkübe edildiğinde kalan aktivite ortalama % 71 olarak elde edilmiştir. Enzim üre ile yüksek oranda inhibe olurken Triton X-100, CaCl2, ZnCl2, KCl, Na2SO3 PMSF ve 2-Merkaptoetanol varlığında çok düşük oranda etkilenmiştir. Bu çalışmada enzimin belirlenen özellikleri bu enzimin endüstriyel kullanımında potansiyel bir tercih sebebi olabilir.A multifunctional endoxylanase producing Bacillus sp. was isolated from soil. Enzyme synthesis occurred at temperatures between 20 °C and 60 °C with an optimum 37 °C. Analysis of the enzyme by SDS-PAGE revealed 4 active enzyme bands, which were estimated to be 66.5 kDa, 80.6 kDa, 95.5 kDa, and 108.4 kDa. The enzyme has a broad temperature range, between 20 and 90 °C, with an optimum at 40 °C; and maximum activity was at pH 6.0. The enzyme showed a gradual decrease in the remaining activity as the pre-incubation temperature increase. Thermostability was not also increased in the presence of Ca2+. An average of 71% of remaining activity observed when the enzyme incubated between pH 3.6 and 10 for 1 h. The enzyme was highly inhibited by urea, and fairly inhibited by Triton X-100, CaCl2, ZnCl2, KCl, Na2SO3 PMSF, and 2-Mercaptoethanol. The properties of the enzyme presented in this study suggest that this enzyme could be a potential industrial interest

Klebsiella Sp. Suşlarında Ceftriaxone Ve İmipenem Dirençlilik Frekansının Saptanması

Bu çalışmada Çukurova Üniversitesi Balcalı Hastanesi kanalizasyon suyundan izole edilen 145 adet Klebsiella sp. suşlarında ceftriaxone, imipenem, tetracycline, streptomycine, ve augmentin antibiyotiklerine karşı gelişen dirençlilik frekansı saptanarak dirençlilik gelişimi ve yayılmasındaki rolü araştırılmıştır. Klebsiella sp. suşlarında antibiyotik direnç oranları ceftriaxone %55.86, streptomycine %57.93, tetracycline %50.33, augmentin %48.27 ve imipenem %7.58 olarak bulunmuştur. Elde edilen veriler, Klebsiella sp. suşlarında dirençliliğin büyük ölçüde çoklu antibiyotik dirençliliği şeklinde olduğunu göstermektedir. Bütün antibiyotiklere dirençli olan suşlar toplam izolatların yaklaşık %44.13’ünü oluştururken bunların %10.93’ü bütün antibiyotiklere, %82.81’i dört antibiyotiğe, %18.75’i üç antibiyotiğe ve %9.37’si aynı anda iki antibiyotiğe dirençlilik göstermektedirler. Klebsiella sp. suçlarındaki antibiyotik dirençliliğinin ceftriaxone için %88.89, streptomycine %79.77 ve tetracycline %79.46 oranında kromozomal kökenli olduğu gözlenmiştir. Augmentin antibiyotiğine hassas suşlarda eliminasyon testleri sonunda dirençlilik gözlenmiştir. Özellikle ethidium bromide ile yaklaşik %33.33 düzeyinde eksizyon gerçekleşmesi izole edilen Klebsiella sp. suçlarının bu antibiyotiğe büyük ölçüde transpozabl elemanlar taşıdığı saptanmıştır. En yüksek eliminasyon düzeyi imipenem antibiyotiği için elde edilirken, bu sonuç imipenem dirençliliğinin önemli oranda plazmid kökenli olduğu saptanmıştır. Kullanılan eliminatörler içerisinde ceftriaxone, streptomycine ve tetracycline için 100 pg/mL konsantrasyonda ethidium bromide, augmentin ve imipenem için 100 pg/mL konsantrasyonda acridin orange daha etkili bulunmuştur

Characterization of Thermostable and Acidophilic Type II Pullulanase from Geobacillus thermoleovorans NP1 and Its Industrial Applications

This research aimed to produce and characterize a thermophilic and an acidic pullulanase and determine the industrial areas of usage of this enzyme. Pullulanase producing Geobacillus thermoleovorans NP1 (NCBI accession number: MF355416) was isolated from soil samples and screened for pullulanase activity on agar medium containing pullulan (pH 7.0) at 55°C. The highest enzyme production occurred at 45°C and pH 7.0 for 12 h. The supernatant enzyme exhibited its maximal activity at 50°C and pH 6.0. It was highly stable between pH 3.0-12.0 for 24 h and 30-100°C for 60 min. The pullulanase was treated with two each concentration of various metal ions, chelators, detergents and inhibitors. The stability of the enzyme was saved at 5 and 10 mM concentrations of Ca2+, Mg2+, Cu2+, Zn2+, and Ba2+. It showed 145% activity with β-mercaptoethanol (%1). In the presence of 5 mM and 10 mM EDTA, NA1 pullulanase exhibited 92% and 101.3% activities, respectively. Each concentration of PMSF (5 and 10 mM) did not affect the enzyme stability. The enzyme activity was measured as 86.3% with 5% SDS. NP1 pullulanase maintained its initial activity with TritonX-100, Tween20 and Tween80. The native NP1 pullulanase is composed of two active subunits of 112 and 107 kDa. The hydrolysis products of pullulan and starch determined by thin layer chromatography analysis are glucose, maltose, maltotriose, and maltodextrin. This shows that NP1 pullulanase is a type II pullulanase (amylopullulanase). According to the wash performance tests, the mix of pullulanase and 1% commercial detergent removed nearly all of the stains (food gravy, chocolate, baby food). In conclusion, type II pullulanase producing by Geobacillus thermoleovorans NP1 is an acidic-neutral, a thermostable enzyme and resistant to some ions, chelators and detergents. Because of its thermo-acidic activity, it can be used in starch (liquefaction/saccharification), food and detergent industry