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Koyun Karaciğerinden Alkol Dehidrogenaz: Saflaştırma, Karakterizasyon, ve Bazı Antibiyotiklerin Etkileri

Year 2017, Volume: 7 Issue: 3, 151 - 160, 30.09.2017

Abstract

Alkol dehidrogenaz (ADH), her bir alt biriminin bir Zn2+ metal içeren katalitik alana ve bir kofaktör bağlama alanına sahip olduğu dimerik bir enzimdir. Bu enzim, alkolü, aldehide dönüştürür. Bu makale koyun karaciğerinden alkol dehidrogenazın saflaştırılması, karakterizasyonu enzimin aktivitesi üzerine bazı antibiyotiklerin in vitro ekilerine odaklanmaktadır. ADH, DEAE-Sephadex A-50 iyon değişim kromatografsi ve Sephadex G-100 jel fltrasyon kromatografsi vasıtasıyla koyun karaciğerinden 0.5 U mg-1 protein spesifk aktivite ve yaklaşık olarak 52.03-kat saflaştırıldı. Enzimin alt birim ve doğal hallerinin molekül kütleleri jel fltrasyon kromatografsi ve SDS-PAGE ile sırasıyla 38.16 kDa ve 80.49 kDa olarak belirlendi. ADH’ın optimum iyonik şiddeti, sıcaklığı ve pH’sı sırasıyla 400 mM, 40°C ve 10.5 idi. Antibiyotiklerin inhibitor etkileri çeşitli konsantrasyonlarda denendi. Kanamisin sülfat, amikasin sülfat, gentamisin, linkomisin ve klindamisin için IC50 değerleri sırasıyla, 43.31, 36.47, 20.38, 18.73 ve 1.31 mM olarak bulundu.


References

  • Akduman B, Uygun M, Aktaş Uygun D, Akgöl S, Denizli A, 2013. Purification of yeast alcohol dehydrogenase by using immobilized metal affinity cryogels. Materials Science and Engineering, C 33; 4842–4848.
  • Alım Z, Beydemir S, 2016. Some Anticancer Agents Act on Human Serum Paraoxonase-1 to Reduce Its Activity. Chem Biol Drug Des, 88 (2); 188-96.
  • Alım Z, Beydemir Ş, 2012. Effects of some anti-neoplastic drugs on sheep liver sorbitol dehydrogenase. Arch. Physiol. and Biochem., 118; 244-252.
  • Aslan HE, Beydemir Ş, 2017. Phenolic compounds: The inhibition effect on polyol pathway enzymes. Chemico-Biological Interactions, 266; 47-55.
  • Batzias GC, Delis GA, Athanasiou, LV, 2005. Clindamycin bioavailability and pharmacokinetics following oral administration of clindamycin hydrochloride capsules in dogs. Vet .J., 170 (3); 339-45.
  • Beydemir Ş, Çiftçi M, Küfrevioğlu Öİ, Büyükokuroğlu ME, 2002. Effects of gentamicine sulfate on enzyme activities of carbonic anhydrase from human erythrocytes in vitro and from rat erythrocytes in vivo. Biol. Pharm. Bull., 25; 966-969.
  • Beydemir Ş, Kulaçoğlu DN, Çiftçi M, Küfrevioğlu Öİ, 2003. The effects of some antibiotics on sheep lens glucose 6-phosphate dehydrogenase in vitro. Eur. J. Ophthalmol., 13; 155-161.
  • Bradford MM, 1976. Rapid and Sensitive Method for Quantitation of Microgram Quantities of Protein Utilizing Principle of Protein-Dye Binding. Analytical Biochemistry, 72 (1-2); 248-254.
  • Cheng F, Hu, T, An Y, Huang J, Xu Y, 2013. Purification and enzymatic characterization of alcohol dehydrogenase from Arabidopsis thaliana. Protein Expression and Purification, 90; 74–77.
  • Çoban TA, Beydemir Ş, Gucin I, Ekinci D, Innocenti A, Vullo D, Supuran CT, 2009. Sildenafil is a strong activator of mammalian carbonic anhydrase isoforms I-XIV. Bioorgan. Med. Chem., 17; 5791-5795.
  • Demir Y, Beydemir S, 2015. Purification, refolding, and characterization of recombinant human paraoxonase-1. Turkish Journal of Chemistry, 39 (4); 764-776.
  • Drauz K, 2012. Enzyme catalysis in organic synthesis: a comprehensive handbook. John Wiley & Sons.
  • Ekinci D, Beydemir Ş., 2009. Effect of some analgesics on paraoxonase-1 purified from human serum. Journal of Enzyme Inhibition and Medicinal Chemistry, 24 (4); 1034-1039.
  • Giguère S, 2013. Lincosamides, Pleuromutilins, and Streptogramins Antimicrobial Therapy in Veterinary Medicine. John Wiley & Sons, Inc.
  • İşgör MM, Beydemir S, 2010. Some cardiovascular therapeutics inhibit paraoxonase 1 (PON1) from human serum. Eur J Pharmacol, 645 (1-3); 135-42.
  • Itoh N, 2014. Use of the anti-Prelog stereospecific alcohol dehydrogenase from Leifsonia and Pseudomonas for producing chiral alcohols. Applied Microbiology and Biotechnology, 98 (9); 3889-3904.
  • Kent A, Turner MA, Sharland M, Heath PT, 2014. Aminoglycoside toxicity in neonates: something to worry about? Expert Rev Anti Infect Ther, 12 (3); 319-31.
  • Kirmair L, Seiler DL, Skerra A, 2015. Stability engineering of the Geobacillus stearothermophilus alcohol dehydrogenase and application for the synthesis of a polyamide 12 precursor. Appl Microbiol Biotechnol, 99 (24); 10501-13.
  • Kotra LP, Haddad J, Mobashery S, 2000. Aminoglycosides: perspectives on mechanisms of action and resistance and strategies to counter resistance. Antimicrob Agents Chemother, 44 (12); 3249-56.
  • Laemmli UK, 1970. Cleavage of Structural Proteins during Assembly of Head of Bacteriophage-T4. Nature, 227(5259); 680.
  • Lee SL, Lee YP, Wu ML, Chi YC, Liu CM, Lai CL, Yin SJ, 2015. Inhibition of human alcohol and aldehyde dehydrogenases by aspirin and salicylate: Assessment of the effects on first-pass metabolism of ethanol. Biochemical Pharmacology, 95 (1); 71-79.
  • Lee YP, Liao JT, Cheng YW, Wu TL, Lee SL, Liu JK, Yin SJ, 2013. Inhibition of human alcohol and aldehyde dehydrogenases by acetaminophen: Assessment of the effects on first-pass metabolism of ethanol. Alcohol, 47 (7); 559-565.
  • Li J, Jiang Z, Wu H, Liang Y, Zhang Y, Liu J, 2010. Enzyme-polysaccharide interaction and its influence on enzyme activity and stability. Carbohydr. Polym., 82; 160-166.
  • Li Y, Ding WX, 2017. Adipose tissue autophagy and homeostasis in alcohol-induced liver injury. Liver Research, 1-9. https://doi.org/10.1016/j.livres.2017.03.004
  • Lin L, Wagner MC, Cocklin R, Kuzma A, Harrington M, Molitoris BA, Goebl MG, 2011. The Antibiotic Gentamicin Inhibits Specific Protein Trafficking Functions of the Arf1/2 Family of GTPases. Antimicrobial Agents and Chemotherapy, 55 (1); 246-254.
  • Manir MM, Kim, JK, Lee BG, Moon SS, 2012. Tea catechins and flavonoids from the leaves of Camellia sinensis inhibit yeast alcohol dehydrogenase. Bioorg. Med. Chem., 20(7); 2376-81.
  • Mert S, Alım Z, İşgör MM, Beydemir Ş, Kasımoğulları R, 2016. The synthesis of novel pyrazole-3,4-dicarboxamides bearing 5-amino-1,3,4-thiadiazole-2-sulfonamide moiety with effective inhibitory activityagainst the isoforms of human cytosolic carbonic anhydrase I and II. Bioorganic Chemistry, 68; 64–71.
  • Raghava S, Gupta MN, 2010. Purification and characterization of an alcohol dehydrogenase with an unusual specificity towards glycerol from Thermus thermophiles. Bioresource Technology, 101; 2554–2557.
  • Raj SB, Ramaswamy S, Plapp BV, 2014. Yeast alcohol dehydrogenase structure and catalysis. Biochemistry, 53 (36); 5791-803.
  • Söyüt H, Beydemir Ş, 2008. Purification and some kinetic properties of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) liver and metal inhibition. Protein and Peptide Letters, 15; 528-535.
  • Söyüt H, Beydemir Ş, 2012. The impact of heavy metals on the activity of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) kidney. Toxıcol. Ind. Health., 28, 296-305.
  • Stickel F, Moreno C, Hampe J, Morgan MY, 2017. The genetics of alcohol dependence and alcohol-related liver disease. Journal of Hepatology, 66; 195–211.
  • Sunderland JR, Tao X, Butrick EE, Keilich LC, Villa CE, Miecznikowski JR, Jain SS, 2016. Investigation of liver alcohol dehydrogenase catalysis using an NADH biomimetic and comparison with a synthetic zinc model complex. Polyhedron, 114; 145–151.
  • Taber RL, 1998. The competitive inhibition of yeast alcohol dehydrogenase by 2,2,2-trifluoroethanol. Biochemical Education, 26 (3); 239-242.
  • Türkeş C, Söyüt H, Beydemir Ş, 2016. In vitro inhibitory effects of palonosetron hydrochloride, bevacizumab and cyclophosphamide on purified paraoxonase-I (hPON1) from human serum. Environmental toxicology and pharmacology, 42; 252-257.
  • Yılmaz H, Ciftci M, Beydemir S, Bakan E, 2002. Purification of glucose 6-phosphate dehydrogenase from chicken erythrocytes. Investigation of some kinetic properties. Preparative Biochemistry and Biotechnology, 32; 287–301
  • Zheng SY, Xu D, Wang HR, Li J, Zhou HM, 1997. Kinetics of irreversible inhibition of yeast alcohol dehydrogenase during modification by 4,4'-dithiodipyridine. International Journal of Biological Macromolecules, 20 (4); 307-313.

Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics

Year 2017, Volume: 7 Issue: 3, 151 - 160, 30.09.2017

Abstract

Alcohol dehydrogenase (ADH) is a dimeric enzyme in which each subunit of the enzyme has a Zn2+ metal-containing catalytic domain and a cofactor-binding domain. This enzyme converts the alcohol to aldehyde. The present article focuses on the purification, characterization and in vitro effects of some antibiotics on alcohol dehydrogenase from sheep liver. ADH was purified with specific activity of 0.5 U/mg proteins and approximately 52.03-fold from sheep liver by DEAE-Sephadex A-50 ion exchange chromatography and gel filtration on Sephadex G-100. The subunit and the natural molecular weights of the enzyme were determined by gel filtration and SDSPAGE 38.16 kDa and 80.49 kDa, respectively. The optimum ionic strenght, temperature and pH of ADH were found 400 mM, 40 °C and 10.5, respectively. The inhibitory effects of the antibiotics were tested at various concentrations. IC50 values for kanamycin sulfate, amikacin sulfate, gentamicin, lincomycin, and clindamycin were found to be 43.31, 36.47, 20.38, 18.73 and 1.31 mM, respectively

References

  • Akduman B, Uygun M, Aktaş Uygun D, Akgöl S, Denizli A, 2013. Purification of yeast alcohol dehydrogenase by using immobilized metal affinity cryogels. Materials Science and Engineering, C 33; 4842–4848.
  • Alım Z, Beydemir S, 2016. Some Anticancer Agents Act on Human Serum Paraoxonase-1 to Reduce Its Activity. Chem Biol Drug Des, 88 (2); 188-96.
  • Alım Z, Beydemir Ş, 2012. Effects of some anti-neoplastic drugs on sheep liver sorbitol dehydrogenase. Arch. Physiol. and Biochem., 118; 244-252.
  • Aslan HE, Beydemir Ş, 2017. Phenolic compounds: The inhibition effect on polyol pathway enzymes. Chemico-Biological Interactions, 266; 47-55.
  • Batzias GC, Delis GA, Athanasiou, LV, 2005. Clindamycin bioavailability and pharmacokinetics following oral administration of clindamycin hydrochloride capsules in dogs. Vet .J., 170 (3); 339-45.
  • Beydemir Ş, Çiftçi M, Küfrevioğlu Öİ, Büyükokuroğlu ME, 2002. Effects of gentamicine sulfate on enzyme activities of carbonic anhydrase from human erythrocytes in vitro and from rat erythrocytes in vivo. Biol. Pharm. Bull., 25; 966-969.
  • Beydemir Ş, Kulaçoğlu DN, Çiftçi M, Küfrevioğlu Öİ, 2003. The effects of some antibiotics on sheep lens glucose 6-phosphate dehydrogenase in vitro. Eur. J. Ophthalmol., 13; 155-161.
  • Bradford MM, 1976. Rapid and Sensitive Method for Quantitation of Microgram Quantities of Protein Utilizing Principle of Protein-Dye Binding. Analytical Biochemistry, 72 (1-2); 248-254.
  • Cheng F, Hu, T, An Y, Huang J, Xu Y, 2013. Purification and enzymatic characterization of alcohol dehydrogenase from Arabidopsis thaliana. Protein Expression and Purification, 90; 74–77.
  • Çoban TA, Beydemir Ş, Gucin I, Ekinci D, Innocenti A, Vullo D, Supuran CT, 2009. Sildenafil is a strong activator of mammalian carbonic anhydrase isoforms I-XIV. Bioorgan. Med. Chem., 17; 5791-5795.
  • Demir Y, Beydemir S, 2015. Purification, refolding, and characterization of recombinant human paraoxonase-1. Turkish Journal of Chemistry, 39 (4); 764-776.
  • Drauz K, 2012. Enzyme catalysis in organic synthesis: a comprehensive handbook. John Wiley & Sons.
  • Ekinci D, Beydemir Ş., 2009. Effect of some analgesics on paraoxonase-1 purified from human serum. Journal of Enzyme Inhibition and Medicinal Chemistry, 24 (4); 1034-1039.
  • Giguère S, 2013. Lincosamides, Pleuromutilins, and Streptogramins Antimicrobial Therapy in Veterinary Medicine. John Wiley & Sons, Inc.
  • İşgör MM, Beydemir S, 2010. Some cardiovascular therapeutics inhibit paraoxonase 1 (PON1) from human serum. Eur J Pharmacol, 645 (1-3); 135-42.
  • Itoh N, 2014. Use of the anti-Prelog stereospecific alcohol dehydrogenase from Leifsonia and Pseudomonas for producing chiral alcohols. Applied Microbiology and Biotechnology, 98 (9); 3889-3904.
  • Kent A, Turner MA, Sharland M, Heath PT, 2014. Aminoglycoside toxicity in neonates: something to worry about? Expert Rev Anti Infect Ther, 12 (3); 319-31.
  • Kirmair L, Seiler DL, Skerra A, 2015. Stability engineering of the Geobacillus stearothermophilus alcohol dehydrogenase and application for the synthesis of a polyamide 12 precursor. Appl Microbiol Biotechnol, 99 (24); 10501-13.
  • Kotra LP, Haddad J, Mobashery S, 2000. Aminoglycosides: perspectives on mechanisms of action and resistance and strategies to counter resistance. Antimicrob Agents Chemother, 44 (12); 3249-56.
  • Laemmli UK, 1970. Cleavage of Structural Proteins during Assembly of Head of Bacteriophage-T4. Nature, 227(5259); 680.
  • Lee SL, Lee YP, Wu ML, Chi YC, Liu CM, Lai CL, Yin SJ, 2015. Inhibition of human alcohol and aldehyde dehydrogenases by aspirin and salicylate: Assessment of the effects on first-pass metabolism of ethanol. Biochemical Pharmacology, 95 (1); 71-79.
  • Lee YP, Liao JT, Cheng YW, Wu TL, Lee SL, Liu JK, Yin SJ, 2013. Inhibition of human alcohol and aldehyde dehydrogenases by acetaminophen: Assessment of the effects on first-pass metabolism of ethanol. Alcohol, 47 (7); 559-565.
  • Li J, Jiang Z, Wu H, Liang Y, Zhang Y, Liu J, 2010. Enzyme-polysaccharide interaction and its influence on enzyme activity and stability. Carbohydr. Polym., 82; 160-166.
  • Li Y, Ding WX, 2017. Adipose tissue autophagy and homeostasis in alcohol-induced liver injury. Liver Research, 1-9. https://doi.org/10.1016/j.livres.2017.03.004
  • Lin L, Wagner MC, Cocklin R, Kuzma A, Harrington M, Molitoris BA, Goebl MG, 2011. The Antibiotic Gentamicin Inhibits Specific Protein Trafficking Functions of the Arf1/2 Family of GTPases. Antimicrobial Agents and Chemotherapy, 55 (1); 246-254.
  • Manir MM, Kim, JK, Lee BG, Moon SS, 2012. Tea catechins and flavonoids from the leaves of Camellia sinensis inhibit yeast alcohol dehydrogenase. Bioorg. Med. Chem., 20(7); 2376-81.
  • Mert S, Alım Z, İşgör MM, Beydemir Ş, Kasımoğulları R, 2016. The synthesis of novel pyrazole-3,4-dicarboxamides bearing 5-amino-1,3,4-thiadiazole-2-sulfonamide moiety with effective inhibitory activityagainst the isoforms of human cytosolic carbonic anhydrase I and II. Bioorganic Chemistry, 68; 64–71.
  • Raghava S, Gupta MN, 2010. Purification and characterization of an alcohol dehydrogenase with an unusual specificity towards glycerol from Thermus thermophiles. Bioresource Technology, 101; 2554–2557.
  • Raj SB, Ramaswamy S, Plapp BV, 2014. Yeast alcohol dehydrogenase structure and catalysis. Biochemistry, 53 (36); 5791-803.
  • Söyüt H, Beydemir Ş, 2008. Purification and some kinetic properties of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) liver and metal inhibition. Protein and Peptide Letters, 15; 528-535.
  • Söyüt H, Beydemir Ş, 2012. The impact of heavy metals on the activity of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) kidney. Toxıcol. Ind. Health., 28, 296-305.
  • Stickel F, Moreno C, Hampe J, Morgan MY, 2017. The genetics of alcohol dependence and alcohol-related liver disease. Journal of Hepatology, 66; 195–211.
  • Sunderland JR, Tao X, Butrick EE, Keilich LC, Villa CE, Miecznikowski JR, Jain SS, 2016. Investigation of liver alcohol dehydrogenase catalysis using an NADH biomimetic and comparison with a synthetic zinc model complex. Polyhedron, 114; 145–151.
  • Taber RL, 1998. The competitive inhibition of yeast alcohol dehydrogenase by 2,2,2-trifluoroethanol. Biochemical Education, 26 (3); 239-242.
  • Türkeş C, Söyüt H, Beydemir Ş, 2016. In vitro inhibitory effects of palonosetron hydrochloride, bevacizumab and cyclophosphamide on purified paraoxonase-I (hPON1) from human serum. Environmental toxicology and pharmacology, 42; 252-257.
  • Yılmaz H, Ciftci M, Beydemir S, Bakan E, 2002. Purification of glucose 6-phosphate dehydrogenase from chicken erythrocytes. Investigation of some kinetic properties. Preparative Biochemistry and Biotechnology, 32; 287–301
  • Zheng SY, Xu D, Wang HR, Li J, Zhou HM, 1997. Kinetics of irreversible inhibition of yeast alcohol dehydrogenase during modification by 4,4'-dithiodipyridine. International Journal of Biological Macromolecules, 20 (4); 307-313.
There are 37 citations in total.

Details

Primary Language English
Journal Section Kimya / Chemistry
Authors

Yeliz Demir This is me

Bülent Şengül This is me

Bülent Ergun This is me

Şükrü Beydemir

Publication Date September 30, 2017
Submission Date June 19, 2017
Acceptance Date July 10, 2017
Published in Issue Year 2017 Volume: 7 Issue: 3

Cite

APA Demir, Y., Şengül, B., Ergun, B., Beydemir, Ş. (2017). Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics. Journal of the Institute of Science and Technology, 7(3), 151-160.
AMA Demir Y, Şengül B, Ergun B, Beydemir Ş. Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics. J. Inst. Sci. and Tech. September 2017;7(3):151-160.
Chicago Demir, Yeliz, Bülent Şengül, Bülent Ergun, and Şükrü Beydemir. “Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics”. Journal of the Institute of Science and Technology 7, no. 3 (September 2017): 151-60.
EndNote Demir Y, Şengül B, Ergun B, Beydemir Ş (September 1, 2017) Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics. Journal of the Institute of Science and Technology 7 3 151–160.
IEEE Y. Demir, B. Şengül, B. Ergun, and Ş. Beydemir, “Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics”, J. Inst. Sci. and Tech., vol. 7, no. 3, pp. 151–160, 2017.
ISNAD Demir, Yeliz et al. “Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics”. Journal of the Institute of Science and Technology 7/3 (September 2017), 151-160.
JAMA Demir Y, Şengül B, Ergun B, Beydemir Ş. Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics. J. Inst. Sci. and Tech. 2017;7:151–160.
MLA Demir, Yeliz et al. “Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics”. Journal of the Institute of Science and Technology, vol. 7, no. 3, 2017, pp. 151-60.
Vancouver Demir Y, Şengül B, Ergun B, Beydemir Ş. Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics. J. Inst. Sci. and Tech. 2017;7(3):151-60.