Compounds from <em>Terminalia mantaly</em> L. (Combretaceae) Stem Bark Exhibit Potent Inhibition against some Pathogenic Yeasts and Enzymes of Metabolic Significance<strong></strong>

Tchuenmogne MAT, Ngouana TK, Gohlke S, et al. Compounds from &lt;em&gt;Terminalia mantaly&lt;/em&gt; L. (Combretaceae) Stem Bark Exhibit Potent Inhibition against some Pathogenic Yeasts and Enzymes of Metabolic Significance&lt;strong&gt;&lt;/strong&gt;. Preprints. 2016.The chemical investigation of the anti-yeast methanol extract from the stem bark of Terminalia mantaly led to the isolation of seven compounds: 3-O-methyl-4-O-&amp;alpha;-rhamnopyranoside ellagic acid (1), 3-O-mehylellagic acid (2), arjungenin or 2,3,19,23-tetrahydroxyolean-12-en-28-o&amp;iuml;c acid (3), arjunglucoside or 2,3,19,23-tetrahydroxyolean-12-en-28-o&amp;iuml;c acid glucopyranoside (4), 2&amp;alpha;,3&amp;alpha;,24-trihydroxyolean-11,13(18)-dien-28-o&amp;iuml;c acid (5), stigmasterol (6), stigmasterol 3-O-&amp;beta;-D-glucopyranoside (7). Their structures were established by means of spectroscopic analysis and comparison with published data. Compounds 1-5 were tested in vitro for activity against three pathogenic yeast isolates, Candida albicans, Candida parapsilosis and Candida krusei. The activity of compounds 1, 2 and 4 were comparable to that of the reference compound fluconazole (MIC values below 32 &amp;micro;g/ml) against the three tested yeast isolates. They were also tested for inhibitory properties against four enzymes of metabolic significance: Glucose-6-Phosphate Deshydrogenase (G6PD), human erythrocyte Carbonic anhydrase I and II (hCA I and hCA II), Glutathione S-transferase (GST). Compound 4 showed highly potent inhibitory property against the four tested enzymes with overall IC50 values below 4 &amp;micro;M and inhibitory constant (Ki) &amp;lt;3 &amp;micro;M.</jats:p

PURIFICATION, CHARACTERIZATION OF GLUTATHIONE S-TRANSFERASE FROM THE GILL TISSUE OF LAKE VAN FISH AND INHIBITION EFFECTS OF SOME METAL IONS AND PESTICIDES

Glutathione-S-Transferases (GSTs) are xenobiotic metabolizing enzymes that protect cells from toxic drugs and environmental electrophiles. Furthermore, these enzymes have effected the regulators of oxidative stress and toxicity [1]. In earlier studies; GST enzymes have been purified and characterized from many living things such as sheep, bird, fish, bacterium, bovine and human. This study proves the purification and characterization of GST enzyme (E.C. 2.5.1.18) from the Gill Tissue of Lake Van Fish by affinity chromatography. Inhibitory effects of some metal ions and pesticides on GST activity were determined with using the CDNB method under in vitro conditions. Purification degree for the purified enzyme was controlled by SDS-PAGE. Optimal pH, optimal ionic strength, optimal temperature and stable pH were determined as 7.3, 120 mM, 35°C, 8.0, respectively. Inhibitory effects of Al3+, Ba2+, B3+ and Se2- metal ions and Oxamyl, Diniconazole, Carbofuran, Tebuconazole and Atrazine pesticides have been examined on the purified GST enzyme. Inhibition graphics have been drawn in order to find the IC50 values of metals and pesticides showing inhibition from Activity % -[Inhibitor] graphs. Consequently, in vitro inhibition rank order was determined as Al3+ > Ba2+ > Se2- > B3+ for metal ions, Carbofuran > Tebuconazole > Oxamyl > Atrazine > Diniconazole for pesticides

Purification of carbonic anhydrase-II from sheep liver and inhibitory effects of some heavy metals on enzyme activity

In this study; sheep carbonic anhydrase-II (SCA-II) (E.C: 4.2.1.1) was purified from sheep liver and in vitro effects of heavy metals on the enzyme was examined. SCA-II isozyme was purified with about 203 purification fold, a specific activity of 2320 EU/mg-protein and a yield of 72 by using Sepharose-4B-L tyrosine-sulfanilamide affinity gel chromatography. Purity of the SCA-II enzyme was verified by SDS-PAGE technique and subunit molecular mass of the enzyme was found as 29 kDa. In addition to this, inhibitory effects of some metal ions on the enzyme were examined. In this study, sheep liver tissue was chosen; because the liver is an organ in which metal wastes of air, water and food are collected and it is easy to obtain the liver tissue. Because of the very important duties of CA enzyme on living beings, the effect of metals on the CA enzyme was investigated

Siirt Tiftik Keçisi Böbrek Dokusundan Karbonik Anhidraz Enziminin Saflaştırılması, Karakterizasyonu ve Bazı Biyokimyasal Özelliklerinin Araştırılması

Carbonic anhydrase (CA) (E.C:4.2.1.1) is a group of metalloenzyme containing zinc widely distributed from the simplest organisms such as bacteria to the most complex organisms such as plants and animals. CA inhibitors are some of the principal drugs used in the management of canine and feline glaucoma. In earlier studies; CA enzymes successfully have been purified and characterized from many living things such as; sheep, chicken, fish, bovine and human. In this study, the CA enzyme has purified from Siirt Mohair Goat kidney tissue with 902.9 EU x mg-1 of specific activity, 50.19% of purification yield and 83.54 of purification folds by using Sepharose-4B-L-tyrosine-sulfonamide affinity column. The purity of the purified enzyme has confirmed by SDS-PAGE. As the characterization of CA enzyme’s in Siirt Mohair Goat kidney has been done; the optimum ionic strength=25mM, the optimum pH=8.5, the optimum temperature=45ºC and the stable pH=7.0 has been determined. Inhibitory effects of Al3+, Ni2+, Cd2+, Pb2+, Ba2+, Zn2+, B3+, Fe3+, Se2+, Ag+ and Co2+ metal ions have been examined on the purified CA enzyme. Inhibition graphics have been drawn in order to find the IC50 values of metals showing inhibition

Effects of some heavy metals on the activities of carbonic anhydrase enzymes from tumorous and non-tumorous human stomach

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

Purification of CA Isoenzymes from Human Cancerous Colon Tissue and Inhibitory Effects of Some Analgesics on Enzyme Activity

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

Purification and Characterization of a-Carbonic Anhydrase II from Sheep Liver and Examining the Inhibition Effect of Kanamycin on Enzyme Activity

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

Carbonic Anyhdrase Enzyme From The Siirt Mohair Goat Liver :Purification, Characterization and Assessment of Enzyme Kinetics Against Metal Toxicity

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 th

Purification of glutathione S-transferase isoenzymes from tumour and nontumour human stomach and inhibitory effects of some heavy metals on enzymes activities

In this study, glutathione S-transferase (GST) enzyme was purified from nontumour and tumour human gastric tissue and in vitro effects of heavy metals on the enzyme were examined. GST was purified 3089 fold with a specific activity of 20 U/mg and a yield of 78% from gastric tumour tissue; and 1185 fold with a specific activity of 5.69 U/mg and a yield of 50% from nontumour tissue by using glutathione-agarose affinity column, respectively. Enzyme purity was verified by SDS-PAGE and subunit molecular mass was calculated around 26 kDa. The molecular weight of the enzyme was calculated as 52 kDa by using Sephadex G-75 gel filtration column. Then, inhibitory effects of metal ions on the enzymes were investigated. Mg2+ and Cd2+ had inhibitory effect on the enzymes activities. Other kinetic properties of the enzymes were also determined

Purification and Characterization of Carbonic Anhydrase from Agrı Balık Lake Trout Gill (Salmo trutta labrax) and Effects of Sulfonamides on Enzyme Activity

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.Agri Ibrahim Cecen University Scientific Research Projects Unit (BAP). Contract Grant Number: BAP 2012/MYO-02