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    Analyzing the Some Biochemical Parameters of Diabetes Mellitus and Obese Patients who Applied to Siirt State Hospital Endocrine Polyclinic and Their Prevalence
    (Amer Inst Physics, 2016) Karagecili, Hasan; Yerlikaya, Emrah; Aydin, Ruken Zeynep
    Obesity and diabetes are major public health problems throughout the World. Obese individuals body mass index (BMI) is >30 kg/m(2). Obesity is characterized by increased waist circumference, total body fat and hyperglycemia. The increased triglyceride and cholesterol level is also shown in obese individuals. The development of obesity is largely due to the consumption of high energy food and sedentary lifestyle. This study was held with the participation of patients applied to Siirt State Hospital endocrine policlinic for treatment. Our aim is to try to determine the biochemical relation and border line of obese and obese+diabetes mellitus patients. Patients and control group lipid profiles were studied in the hospital biochemisty laboratory. Laboratory results of diabetes+obese, obese and control groups were evaluated. Patients and control samples blood serum levels were compared according to their lipid profiles. In 2015, 735 diabetes mellitus type 2 patients applied to Endocrine polyclinic. Some of these patient's serum levels were evaluated. Difference between diabetes+obese and diabetes groups were near critical level for LDL and trigliserid. There were not observed statistically significant difference between groups in terms of HDL and cholesterol. There were found significant difference between groups for blood glucose p<0.003, age p<0.001. According to gender between women and men serum levels, ALT and AST levels; p<0.006 and cholesterol; p<0.04 were detected. According to participants education level blood biochemistry levels were observed statisticaly different p<0.001 with non-literacy group. In conclusion, obese and obese+diabetes patients blood serum values nearly close to each other. Obese subjects were been diabetic obese with age. In women obesity and diabetes mellitus prevalence were seen too much.
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    Bazı yem ve gübre sanayi kimyasallarının hyalüronidaz üzerine in vitro etkileri
    (2016) Kaya, Mustafa Oğuzhan; Yerlikaya, Emrah; Özyazıc, Mehmet Arif; İrak, Kıvanç
    ABSTRACT This study investigates in vitro effects of some chemicals, used as a spreading factor on therapeutic and various medical fields and applied frequently in forage and fertilizer industry on bovine testicular hyaluronidase (BTH) which is substantially effective in glucotechnological applications and delivery of artificial insemination. BTH was purified through ammonium sulfate precipitation method and affinity gel (Sepharose-4B-L- tyrosine -m-anisidine). The inhibiton effects of Fe2SO4, C2H4N4, K2SiF6, CH2N2, CH4N2O and indole-butyric acid were determined on purified BTH. IC50 values of these chemicals were found as 7.097, 5.775, 6.854, 5.317, 4.617, 4.265 ×10-2 mM respectively and at the end of research, indole-butyric acid (IBA) was found as the strongest inhibitory effect compound by the value of IC50 with 4.265×10-2 mM.
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    CARBONIC ANHYDRASE ENZYME FROM THE Alectoris chukar LIVER PURIFICATION CHARACTERIZATION AND ASSESSMENT OF ENZYME KINETICS AGAINST SOME PESTICIDES AND METAL TOXICITY
    (Parlar Scientific Publications (P S P), 2021) Karagecili, Hasan; Demirdag, Ramazan; Yerlikaya, Emrah
    Carbonic anhydrases (CAs) take major roles in many pathological situations associated with convulsions, carcinoma, glaucoma, weight problems, etc. In former researches; using tissues and erythrocytes of a wide range of organisms like; dogs, swine, sheep, chickens, bees, fish, goats, bovines, bacteria, and humans, CA enzymes have been purified and characterized. With this research, the CA enzyme has purified from Alectoris chukar liver tissue with 1.23 EU x mg(-1) of specific activity, 55.5% purification yield, and 460.7 purification folds. The purity of the purified enzyme has been approved by SDS-PAGE. The molecular weight was estimated to be about 28.4 kDa. As the characterization of CA enzyme's in the Alectoris chukar liver has been done; the optimum ionic strength-50 mM Tris-SO4. the optimum pH-8.0, and the optimum temperature-35 degrees C have been decided. K-M and V-max values were calculated by Lineweaver Burk graph. K-M constants were estimated as 1.035 mM, V-max, values as 0.02 molxmin(-1) for 4-nitrophenyl acetate. Inhibitory consequences of some pesticides (Spinosad and Dimethoate) and metal ions (Al3+, Cu2+, Ba2+, Fe2+, Mn2+, Se2+) have been analyzed on the purified CA enzyme. Inhibition graphics were drawn to find the IC50 values of metals and pesticides displaying inhibition.
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    Carbonic Anyhdrase Enzyme From The Siirt Mohair Goat Liver : Purification, Characterization and Assessment of Enzyme Kinetics Against Metal Toxicity
    (2017) Yerlikaya, Emrah; Karageçili, Hasan; Demirdağ, Ramazan; Kaya, Mustafa Oğuzhan
    Because of their physiological and clinical roles, carbonic anhydrases (CAs) are the most studied enzymes. In earlier studies; CA enzymes have been purified and characterized from the tissues and erythrocytes of many organisms such as; dog, swine, sheep, chicken, bee, fish, bovine, bacteria and human. In this study, the CA enzyme has purified from Siirt Mohair Goat liver tissue with 1930.84 EU.mg-1 of specific activity, 57.28% of purification yield and 80.55 of purification folds. The purity of the purified enzyme has confirmed by SDS-PAGE. As the characterization of CA enzyme’s in Siirt Mohair Goat liver has been done; the optimum ionic strength: 25 mM, the optimum pH: 8.0, the optimum temperature: 40ºC and the stable pH: 7.0 has been determined. Inhibitory effects of some metal ions have been examined on the purified CA enzyme. IC50 values of inhibiting metal ions were found as 2.24, 2.76, 2.36, 3.20, 2.55, 2.25, 3.28, 2.13, 3.10, 1.75, 2.16 and 3.50 mM for Al3+, Ni2+, Cd2+, Cu2+, Pb2+, Ba2+, Zn2+, B3+, Fe3+, Se2+, Ag+ and Co2+ respectively. As a result, CA enzyme was first purified from the Siirt Mohair Goat liver and the characteristics of the enzyme were investigated in this study.
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    Carbonic Anyhdrase Enzyme From The Siirt Mohair Goat Liver :Purification, Characterization and Assessment of Enzyme Kinetics Against Metal Toxicity
    (2017) Yerlikaya, Emrah; Karagecili, Hasan; Demirdag, Ramazan; Kaya, Mustafa Oguzhan
    Because of their physiological and clinical roles, carbonic anhydrases (CAs) are the most studied enzymes. In earlier studies; CA enzymes have been purified and characterized from the tissues and erythrocytes of many organisms such as; dog, swine, sheep, chicken, bee, fish, bovine, bacteria and human. In this study, the CA enzyme has purified from Siirt Mohair Goat liver tissue with 1930.84 EU x mg-1 of specific activity, 57.28% of purification yield and 80.55 of purification folds. The purity of the purified enzyme has confirmed by SDS-PAGE. As the characterization of CA enzyme’s in Siirt Mohair Goat liver has been done; the optimum ionic strength=25 mM, the optimum pH= 8.0, the optimum temperature= 40ºC and the stable pH= 7.0 has been determined. Inhibitory effects of some metal ions have been examined on the purified CA enzyme. IC50 values of inhibiting metal ions were found as 2.24, 2.76, 2.36, 3.20, 2.55, 2.25, 3.28, 2.13, 3.10, 1.75, 2.16 and 3.50 mM for Al3+, Ni2+, Cd2+, Cu2+, Pb2+, Ba2+, Zn2+, B3+, Fe3+, Se2+, Ag+ and Co2+ respectively. As a result, CA enzyme was first purified from the
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    Comparison of biochemical parameters of prevalent hemoglobinopathies with healthy individuals
    (2020) Yerlikaya, Emrah; Karageçili, Hasan; Kaya, Mustafa Oğuzhan
    Thalassemia is the most frequently seen monogenetic disorders around the world that is inherited as a recessive single-gene disease, resulting from mutations in ?-or?-globin gene clusters. In this study Beta Thalassemia Intermedia, Beta Thalassemia Minor, Sickle Cell Disease patients and the healthy control individuals both incidence and serum parameters were examined and analyzed retrospectively between 2015-2017 from Public Health information system. Beta Thalassemia Intermedia, BetaThalassemia Major, Beta Thalassemia Minor and Sickle Cell Disease patient groups incidences were evaluated on the basis of their HPLC analysis. Patients and controlgroups serum parameters levels were analyzed from their autoanalyzer results. These serum biochemical tests datas were controlled and taken from Siirt Public HealthCentre information system. In each year, there were recorded approximately 12 Beta Talassemia patients, 2 Beta Thallassemia Major patients, 340 Beta ThalassemiaMinor patients and 33 Sickle Cell Disease patients.Beta Thalassemia Intermedia patients serum parameters levels were compared with Beta Thalassemia Minor Patientsserum parameters and control group serum levels. There were seen statisticaly significant difference between Beta Thalassemia Intermedia patient’s serum ast, albumin,creatine, amylase, phosphor, iron, iron binding capacity, total protein, sodium, bilirubin direct, bilirubin total, LDH level and control serum parameters levels, p<0.05. Butthere were not seen statistically significant differences between Beta Thalassemia Minor patients serum levels and control serum levels. Studies like this should be donenationwide, in each city to determine the current state of abnormal hemoglobinopathies in Turkey. Thus, a broader, comprehensive national screening program is needed.
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    Effects of some heavy metals on the activities of carbonic anhydrase enzymes from tumorous and non-tumorous human stomach
    (2015) Yerlikaya, Emrah; Demirdag, Ramazan; Kufrevioglu, Omer Irfan; Gundogdu, Cemal
    In this study, in vitro effects of certain heavy metals on the human carbonic anhydrase enzyme were examined. Inhibitory effects of metal ions (Pb2+, Cu2+, Fe2+, Cr2+, Al3+, Ni2+, Mn2+, Cd2+, Zn2+, and Mg2+) were observed in tumorous and non-tumorous tissue. IC50 values were calculated for metals. The Cu2+, Zn2+, Ni2+, Cd2+ and Mg2+ IC50 values of tumorous tissue were calculated as 0.034 mM, 0.426 mM, 0.597 mM, 0.878 mM and 2.52 mM, respectively. The Cu2+, Zn2+, Ni2+, Cd2+ and Mg2+ IC50 values of non-tumorous tissue were calculated as 0.067 mM, 0.991 mM, 1.065 mM, 1.724 mM and 6.13 mM, respectively. Carbonic anhydrase activity was measured as described by Wilbur and Anderson. Hydratase activity was used to determine IC50 values. In this study, tumorous and non-tumorous human stomach tissues were selected due to the fact that among the diseases, stomach cancer has one of the highest mortality rates. Stomach cancer, a type of cancer affecting the digestive system, is a fatal disease in living systems. The effects of metals on the CA enzyme were investigated due to the extremely important role that CA enzymes play in living beings.
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    Examination of Changes in Enzyme Activities of Erythrocyte Glucose 6-Phosphate Dehydrogenase and 6-Phosphogluconate Dehydrogenase in Rats Given Naringenin and Lead Acetate
    (2015) Demirdag, Ramazan; Comakli, Veysel; Ozkaya, Ahmet; Sahin, Zafer; Dag, Uzeyir; Yerlikaya, Emrah; Kuzu, Muslum
    In our study, controlled experimental groups were performed by giving substances Lead acetate, Naringenin andNaringenin+Lead acetate to rats in vivo conditions Changes in the glucose 6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) enzyme activities in erythrocytes of rats in these groups were compared to the Control group. An inhibition significant degree for G6PD enzyme activity was observed in all groups when compared to the Control group (p < 0.001). While inhibition significant degree for 6PGD enzyme activity was observed in Lead acetate groups (p < 0.001), no significant effect was observed in the Naringenin and Naringenin + Lead acetate groups (p > 0.05). In addition, lead levels in the groups of rats were determined using an inductively coupled plasma mass spectrometer (ICP-MS) device. As a result of measurements by the ICP-MS device, lead levels were found as an average of 42.9 ± 2.51, 36.71 ± 1.13, 172.16 ± 9.63, and 95.07 ± 5.87 ppm in the Control, Naringenin, Lead acetate and Naringenin+Lead acetate groups, respectively. Our results were shown thatNaringenin has protective effects on the Lead acetate induced oxidative stress erythrocytes in rat.
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    Expression of hCA IX isoenzyme by using sumo fusion partner and examining the effects of antitumor drugs
    (2015) Yerlikaya, Emrah; Erdogan, Orhan; Demirdag, Ramazan; Senturk, Murat; Kufrevioglu, Omer Irfan
    Objective: In this study, investigating the effects of inhibition of the enzyme activity of some antitumor drugs and the Cancer-Related Human Carbonic Anhydrase IX (hCA IX) isoenzyme expressing as a SUMO fusion protein in an Escherichia coli expression system were aimed. Methods: hCA IX isoenzyme was expressed using SUMO fusion technology. The fusion protein was expressed in a totally soluble form and the expression was verified by SDS-PAGE analysis. Affinity chromatography was used in the purification processes. The effects of certain antitumor drugs on enzyme activity were investigated in vitro conditions by using esterase activity. IC50 values of drugs showing the inhibitory effect were calculated. Inhibition types and Ki values for antitumor drugs, which inhibit the enzyme, were determined by separately plotting Lineweaver- Burk plots. Results: The molecular weight of the fusion protein was approximately 85kDa. The optimal induction concentration of IPTG and the growth temperature were found to be 1.0mM and 30oC. The fusion protein was purified at approximately 3.07-fold with a yield of 92.58%, and a specific activity of 43707EU/mg proteins by nickel nitrilo-triacetic acid resin chromatography.
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    First remarkable findings in comparison of patients in Siirt / Turkey in novel coronavirus(Covid-19) pandemic
    (2020) Alayunt, Naci Ömer; Özüdoğru, Osman; Yerlikaya, Emrah
    In December 2019, a novel coronavirus (Covid-19) associated with human to human transmission and severe human infection has been recently reported from the city of Wuhan in China Hubei Province. The first Covid-19 in Turkey comparative findings of their patients can help spread the infection, early diagnosis, treatment and control. This study in Turkey Siirt State Hospital in Covid-19 diagnosed patients and 12 healthy normal consists of 12 patients. Data from electronic medical records were taken retrospectively and patient and control group data were compared statistically. Statistically decreased lymphocyte (1.23 ± 0.66), WBC (5.00 ± 1.50), neutrophil (3.32 ± 1.77), platelets (175.08 ± 44.18) in Covid-19 patients compared to the control group and increased D-Dimer (595.08 ± 185.87), ferritin (236.31 ± 50.36), CRP (12.43 ± 4.31), lactate (1.37 ± 0.39), CK (189.50 ± 63.66) levels, other clinical findings and CT images were found to be compatible with Covid-19. This study is the first comparative study on pandemic Covid-19 in Siirt/Turkey. These prognostic parameters are important for a specific clinical diagnosis, which may contribute to reflect the evolution of CT findings and the severe course of the disease and the worst clinical picture.
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    Heavy metal ion inhibition studies of human, sheep and fish ?-carbonic anhydrases
    (2013) Demirdag, Ramazan; Yerlikaya, Emrah; Senturk, Murat; Kufrevioglu, Omer Irfan; Supuran, Cladiu T.
    Carbonic anhydrases (CAs, EC 4.2.1.1) were purified from sheep kidney (sCA IV), from the liver of the teleost fish Dicentrarchus labrax (dCA) and from human erythrocytes (hCA I and hCA II). The purification procedure consisted of a single step affinity chromatography on Sepharose 4B-tyrosine-sulfanilamide. The kinetic parameters of these enzymes were determined for their esterase activity with 4-nitrophenyl acetate as substrate. The following metal ions, Pb2+, Co2+, Hg2+, Cd2+, Zn2+, Se2+, Cu2+, Al3+ and Mn3+ showed inhibitory effects on these enzymes. The tested metal ions inhibited these CAs competitively in the low milimolar/submillimolar range. The susceptibility to various cations inhibitors differs significantly between these vertebrate ?-CAs and is probably due to their binding to His64 or the histidine cluster.
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    Influence of pesticides on the pH regulatory enzyme, carbonic anhydrase, from European Seabass liver and bovine erythrocytes
    (2012) Demirdag, Ramazan; Yerlikaya, Emrah; Aksakal, Ercüment; Kufrevioglu, Omer Irfan; Ekinci, Deniz
    The objective of this study was to assess the inhibitory effects of six commonly used pesticides, cyhalothrin, cypermethrin, dichlorvos, methamidophos, chlorpyrifos and methylparathion, on the pH regulatory enzyme carbonic anhydrase (CA) of Dicentrarchus labrax (European Seabass) liver (dCA) and bovine erythrocytes (bCA). Results of the study showed that the pesticides displayed quite variable inhibition profiles with KI values ranging from 0.376 to 26.164 M against dCA, and from 1.174 to 53.281 M against bCA. Methyl- parathion was the most effective inhibitor for both enzymes. Overall data show that all of the tested pesticides inhibit both dCA and bCA at low concentrations indicating that indiscriminate use of these pesticides might cause disruption of acid base regulation resulting in animal deaths. Our results also point out that susceptibility to these pesticides varies among CAs from different organisms.
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    Inhibition Effects of Some Sulfonamides on Carbonic Anhydrase from European seabass (Dicentrachus labrax)
    (2011) Erdem, Deryanur; Yerlikaya, Emrah; Ceyhun, Saltuk Bugrahan; Demirdag, Ramazan; Senturk, Murat; Erdogan, Orhan; Kufrevioglu, Omer Irfan
    Alpha-carbonic anhydrase (EC: 4.2.1.1; CA) was purified from European seabass gill and liver. The purification procedure was composed of preparation of homogenate (or hemolysate) and affinity chromatography on Sepharose 4B-tyrosine-sulfanilamide. Some sulfonamides exhibited in vitro inhibitory effects on the enzyme activity. Ki constants and IC50 values for these drugs were determined by Lineweaver-Burk graphs and plotting activity % vs. [I], respectively. IC50 values of sulfanilamide, mafenide, acetazolamide, 2-amino-1,3,5-tiyadiazol-5 sulfonamide were 980 ??M, 142 ??M, 20 ??M and 34 ??M for gill carbonic anhydrase (GCA), 126 ??M, 23 ??M, 14 ??M and 2.58 ??M for liver carbonic anhydrase (LCA), respectively. Some sulfonamides exhibited much stronger inhibitory activity against GCA and LCA at low micromolar concentrations with the Ki values ranging from 0.21 to 76.0 ??M as compared with other CAs.
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    INHIBITORY EFFECTS OF SOME PESTICIDES AND METALS ON CARBONIC ANHYDRASE PURIFIED FROM SHABUT FISH BARBUS GRYPUS ) GILL TISSUE
    (2019) Yerlikaya, Emrah
    Carbonic anhydrase enzyme catalyzes the reversible inter-conversion of CO2 and HCO3. The enzyme is crucial for the osmotic balance and acid–base regulation in the fish. It is well-known that gills of fish play the most important role in acid–base relevant ion transfer, the transfer of H+ and/or HCO3, for the maintenance of systemic pH. Many researches have shown that fish are the species that is the most susceptible to environmental toxins. In addition, these toxins firstly encounter the gill tissue in fish. In this study, the carbonic anhydrase enzyme was purified 198.6-folds with 58.8% yield from Shabut Fish (Barbus grypus) gill tissue by Sepharose-4B-L-tyrosine-sulfanilamide affinity column. The specific activity was determined as 2.92 EU/mg protein. The molecular weight determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis was found to be about 29.9 kDa. Inhibitory effects of some pesticides (Spinosad and Dimethoate) and metal ions (Al3+, Cu2+, Ba2+, Fe2+, Mn2+, Se2+) were examined on the purified carbonic anhydrase enzyme. Inhibition graphics were drawn in order to find the IC50 values of metals and pesticides showing inhibition. The kinetic parameters of this enzyme were determined for its esterase activity, with 4-nitrophenyl acetate as substrate.
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    Investigation of Changes in Nutrition/Eating Habits and Physical Activity Status of Nursing Students in the COVID-19 Period and Determination of Vaccination Status
    (2022) Yerlikaya, Emrah; Çelik, Halil Coşkun; Karageçili, Hasan
    In this study, it was aimed to examine the changes in the nutrition/eating habits and physical activity status of nursing students during the coronavirus disease 2019 (COVID-19) period and to determine the vaccination status of the students. In this descriptive cross-sectional study, 324 students studying at the Faculty of Health Sciences Nursing Department of a university and selected by random sampling method were included. Descriptive statistics (frequency, percentage) and chi-square test were used in the analysis of the data collected through the online survey. Due to the COVID-19 pandemic, an online questionnaire was prepared in the \"Google form\" application and students were provided to fill it voluntarily. It was determined that 123 (31.8%) of the students had COVID-19. It was determined that there were 258 (79.6%) students who were voluntarily vaccinated, 56 (17.3%) vaccinated against their own will, and 10 (3.1%) students who were not vaccinated. 276 (85.2%) students with BioNTech vaccine and 38 (11.7%) students with Sinovac vaccine were identified. 246 (75.9%) students who received two doses of vaccine, 40 (12.3%) students who received one dose of vaccine and 28 (8.6%) students who received three doses of vaccine were identified. When male and female students were compared based on body mass index in terms of maintaining their body weight, weight gain was observed in males. Although there was a decrease in physical activities and an increase in insomnia problems during the pandemic period, there was no increase in the rates of smoking and alcohol use.
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    Purification and Characterization of a-Carbonic Anhydrase II from Sheep Liver and Examining the Inhibition Effect of Kanamycin on Enzyme Activity
    (2012) Comakli, Veysel; Yerlikaya, Emrah; Demirdag, Ramazan; Kufrevioglu, Omer Irfan
    Sheep carbonic anhydrase - II (SCA-II) (E.C: 4.2.1.1) was purified from sheep liver and some characteristic properties were investigated. The enzyme was purified approximate 43.1-fold with a yield of 38.6%, and a specific activity of 4000 EU/mg proteins. For the enzyme, optimum pH, optimum temperature, optimum ionic strength and stable pH were determined to be 7.5, 40 ºC, 10 mM and 8.5, respectively. The molecular weight was found 29 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Kanamycin exhibited in vitro inhibitory effect on the enzyme activity.
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    Purification and Characterization of Carbonic Anhydrase from A. gri Balik Lake Trout Gill (Salmo trutta labrax) and Effects of Sulfonamides on Enzyme Activity
    (Wiley, 2015) Demirdag, Ramazan; Comakli, Veysal; Kuzu, Muslum; Yerlikaya, Emrah; Senturk, Murat
    Carbonic anhydrase (CA) was purified from Ar Balk Lake trout gill (fCA) by affinity chromatography on a sepharose 4B-tyrosine-sulfanilamide column. The fCA enzyme was purified with about a 303.9 purification factor, a specific activity 4130.4 EU (mg-protein)(-1), and a yield of 79.3 by using sepharose-4B-l tyrosine-sulfanilamide affinity gel chromatography. The molecular weight determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was found to be about 29.9 kDa. The kinetic parameters, K-M and V-max were determined for the 4-nitrophenyl acetate hydrolysis reaction. Some sulfonamides were tested as inhibitors against the purified CA enzymes. The K-i constants for mafenide (1), p-toluenesulfonamide (2), 2-bromo-benzene sulfonamide (3), 4-chlorobenzene sulfonamide (4), 4-amino-6-chloro-1-3 benzenedisulfonamide (5), sulfamethazine (6), sulfaguanidine (7), sulfadiazine (8), and acetozazolamide (9) were in the range of 7.5-108.75 M. (C) 2014 Wiley Periodicals, Inc.
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    Purification and Characterization of Carbonic Anhydrase from Agrı Balık Lake Trout Gill (Salmo trutta labrax) and Effects of Sulfonamides on Enzyme Activity
    (2015) Demirdag, Ramazan; Comakli, Veysel; Kuzu, Muslum; Yerlikaya, Emrah; Senturk, Murat
    Carbonic anhydrase (CA) was purified from Agrı Balık Lake trout gill (fCA) by affinity chromatography on a sepharose 4B-tyrosine-sulfanilamide column. The fCA enzyme was purified with about a 303.9 purification factor, a specific activity 4130.4 EU (mg-protein)–1, and a yield of 79.3 by using sepharose- 4B-L tyrosine-sulfanilamide affinity gel chromatography. The molecular weight determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) was found to be about 29.9 kDa. The kinetic parameters, KM and Vmax were determined for the 4-nitrophenyl acetate hydrolysis reaction. Some sulfonamides were tested as inhibitors against the purified CA enzymes. The Ki constants for mafenide (1), p-toluenesulfonamide (2), 2-bromo-benzene sulfonamide (3), 4-chlorobenzene sulfonamide (4), 4-amino-6- chloro-1–3 benzenedisulfonamide (5), sulfamethazine (6), sulfaguanidine (7), sulfadiazine (8), and acetozazolamide (9) were in the range of 7.5–108.75 µM.
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    Purification and characterization of carbonic anhydrase from the teleost fish Dicentrarchus labrax (European seabass) liver and toxicological effects of metals on enzyme activity
    (2011) Ceyhun, Saltuk Bugrahan; Senturk, Murat; Yerlikaya, Emrah; Erdogan, Orhan; Kufrevioglu, Omer Irfan; Ekinci, Deniz
    Carbonic anhydrase (EC 4.2.1.1; CA) was purified and characterized from the liver of the teleost fish Dicentrarchus labrax (European seabass) for the first time. The purification procedure consisted of a single step affinity chromatography on Sepharose 4B-tyrosinesulfanilamide. The enzyme was purified 78.8-fold with a yield of 46%, and a specific activity of 751.72U/mg proteins. It has an optimum pH at 7.5; an optimum temperature at 25 ?C; an optimum ionic strength at 10mM and a stable pH at 8.5. The kinetic parameters of this enzyme were determined for its esterase activity, with 4-nitrophenyl acetate (NPA) as substrate and the purified enzyme had an apparent KM and Vmax values of 0.44mM and 0.249 mol x min-1, respectively. The following metals, Al+3, Cu+2, Pb+2, Co+3, Ag+1, Zn+2 and Hg+2 showed inhibitory effects on the enzyme. Al+3 and Cu+2 exhibited the strongest inhibitory action. Pb+2 was moderate inhibitor, whereas other metals showed weaker actions. All tested metals inhibited the enzyme in a competitive manner. Our findings indicate that these metals inhibit the fish enzyme in a similar manner to other -CAs from mammals investigated earlier, but the susceptibility to various metals differ between the fish and mammalian enzymes. Our results also demonstrate that these metals might be dangerous at low micromolar concentrations for fish CA enzymes.
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    Purification of CA Isoenzymes from Human Cancerous Colon Tissue and Inhibitory Effects of Some Analgesics on Enzyme Activity
    (2012) Demirdag, Ramazan; Yerlikaya, Emrah; Kufrevioglu, Omer Irfan; Gundogdu, Cemal
    Carbonic Anhydrase (CA) is an enzyme which is responsible for the hydration of carbon dioxide to carbonic acid and it also takes places in many biological processes in the living organisms. In this study, CA isoenzymes (CA II and CA IX) together were purified 78.4 fold with a yield of 54.86 and specific activity of 106.67 by using Sepharose 4B-L-tyrosine sulfanilamide affinity chromatography. In SDS-PAGE molecular weights of CA II and CA IX were calculated as 29 kDa and 56 kDa respectively. Besides inhibitory effects of some analgesics on purified total enzyme was investigated. IC50 values were found as 0.0077, 0.025, 0.011 and 0.04 mM for dexketoprofen, pethidine, phenyramidol and tramadol respectively.