Inhibition effects of some pesticides and heavy metals on carbonic anhydrase enzyme activity purified from horse mackerel (Trachurus trachurus) gill tissues

  • Cuneyt CaglayanEmail author
  • Parham Taslimi
  • Cebrahil Türk
  • İlhami Gulcin
  • Fatih Mehmet Kandemir
  • Yeliz Demir
  • Şükrü Beydemir
Research Article


The gill tissue is the main site of metabolic enzymes or compensation, with the kidney tissue playing a supporting role. At the gill tissue, carbonic anhydrase enzymes (CAs) catalyze the hydration of CO2 to HCO3 and H+ for production to the H2O. In this work, the CA enzyme was purified from horse mackerel (Trachurus trachurus) gill with a specific activity of 21,381.42 EU/mg, purification fold of 150.61, total activity of 2347.68 EU/mL, and a yield of 16.13% using sepharose 4B-L-tyrosine-sulfanilamide affinity gel chromatography. For recording the enzyme purity, gel electrophoresis was performed, and single band was seen. The molecular weight of this enzyme was found approximately 35 kDa. Also, the inhibitory effects of different pesticides such as thiram, clofentezine, propineb, deltamethrin, azoxystrobin, and thiophanate and heavy metal ions such as Fe2+, Cu2+, Co2+, Pb2+ Hg2+, and As3+ on horse mackerel gill tissue CA enzyme activities were investigated. Our results indicated that these pesticides and metal ions showed inhibitory effects at low nanomolar and millimolar concentrations for fish gill CA enzymes, respectively.


Carbonic anhydrase Horse mackerel Trachurus trachurus Gill Heavy metal Pesticides 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. Abbaci A, Azzouz N, Bouznit Y (2014) A new copper doped montmorillonite modified carbon paste electrode for propineb detection. Appl Clay Sci 90:130–134CrossRefGoogle Scholar
  2. Aslan HE, Demir Y, Özaslan MS, Türkan F, Beydemir Ş, Küfrevioğlu ÖI (2018) The behavior of some chalcones on acetylcholinesterase and carbonic anhydrase activity. Drug Chem Toxicol:1–7Google Scholar
  3. Atmaca U, Kaya R, Karaman HS, Çelik M, Gülçin İ (2019) Synthesis of oxazolidinone from enantiomerically enriched allylic alcohols and determination of their molecular docking and biologic activities. Bioorg Chem 88:102980CrossRefGoogle Scholar
  4. Bayindir S, Caglayan C, Karaman M, Gülcin İ (2019) The green synthesis and molecular docking of novel N-substituted rhodanines as effective inhibitors for carbonic anhydrase and acetylcholinesterase enzymes. Bioorg Chem 103096Google Scholar
  5. Bayrak Ç, Taslimi P, Gülçin İ, Menzek A (2017) The first synthesis of 4-phenylbutenone derivative bromophenols including natural products and their inhibition profiles for carbonic anhydrase, acetylcholinesterase and butyrylcholinesterase enzymes. Bioorg Chem 72:359–366CrossRefGoogle Scholar
  6. Beydemir Ş, Demir Y (2017) Antiepileptic drugs: impacts on human serum paraoxonase-1. J Biochem Mol Toxicol 31:e21889CrossRefGoogle Scholar
  7. Boztas M, Cetinkaya Y, Topal M, Il G, Menzek A, Sahin E, Tanc M, Supuran CT (2014) Synthesis and carbonic anhydrase isoenzymes I, II, IX, and XII inhibitory effects of dimethoxybromophenol derivatives incorporating cyclopropane moieties. J Med Chem 58:640–650CrossRefGoogle Scholar
  8. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. AnBio 72:248–254Google Scholar
  9. Burmaoglu S, Yilmaz AO, Polat MF, Kaya R, Gulcin İ, Algul O (2019) Synthesis and biological evaluation of novel tris-chalcones as potent carbonic anhydrase, acetylcholinesterase, butyrylcholinesterase and α-glycosidase inhibitors. Bioorg Chem 85:191–197CrossRefGoogle Scholar
  10. Bustaffa E, Stoccoro A, Bianchi F, Migliore L (2014) Genotoxic and epigenetic mechanisms in arsenic carcinogenicity. Arch Toxicol 88:1043–1067CrossRefGoogle Scholar
  11. Butler S, Kelly H, Mueller T, Kruger G, Cochran A, Raper T (2018) Influence of droplet size and azoxystrobin insensitivity on frogeye leaf spot management in soybean. Crop Protect 112:149–158CrossRefGoogle Scholar
  12. Caglayan C (2019) The effects of naringin on different cyclophosphamide-induced organ toxicities in rats: investigation of changes in some metabolic enzyme activities. Environ Sci Pollut Res:1–10Google Scholar
  13. Caglayan C, Gulcin İ (2018) The toxicological effects of some avermectins on goat liver carbonic anhydrase enzyme. J Biochem Mol Toxicol 32:e22010CrossRefGoogle Scholar
  14. Caglayan C, Demir Y, Kucukler S, Taslimi P, Kandemir FM, Gulçin İ (2019a) The effects of hesperidin on sodium arsenite-induced different organ toxicity in rats on metabolic enzymes as antidiabetic and anticholinergics potentials: a biochemical approach. J Food Biochem 43:e12720CrossRefGoogle Scholar
  15. Caglayan C, Kandemir FM, Yildirim S, Kucukler S, Eser G (2019b) Rutin protects mercuric chloride-induced nephrotoxicity via targeting of aquaporin 1 level, oxidative stress, apoptosis and inflammation in rats. J Trace Elem Med Biol 54:69–78CrossRefGoogle Scholar
  16. Ceyhun SB, Şentürk M, Ekinci D, Erdoğan O, Çiltaş A, Kocaman EM (2010) Deltamethrin attenuates antioxidant defense system and induces the expression of heat shock protein 70 in rainbow trout. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 152:215–223Google Scholar
  17. Ceyhun SB, Sentürk M, Yerlikaya E, Erdoğan O, Küfrevioğlu Oİ, Ekinci D (2011) Purification and characterization of carbonic anhydrase from the teleost fish Dicentrarchus labrax (European seabass) liver and toxicological effects of metals on enzyme activity. Environ Toxicol Pharmacol 32(1):69–74CrossRefGoogle Scholar
  18. Demir Y, Beydemir Ş (2015) Purification, refolding, and characterization of recombinant human paraoxonase-1. Turk J Chem 39:764–776CrossRefGoogle Scholar
  19. Demir Y, Oruç E, Topal A (2016) Carbonic anhydrase activity responses and histopathological changes in gill and liver tissues after acute exposure to chromium in brown trout juveniles. Hacettepe J Biol Chem 44:515–523Google Scholar
  20. Demir Y, Şenol Kotan M, Dikbaş N, Beydemir Ş (2017) Phytase from Weissella halotolerans: purification, partial characterisation and the effect of some metals. Int J Food Prop 20:2127–2137Google Scholar
  21. Erdemir F, Celepci DB, Aktaş A, Gök Y, Kaya R, Taslimi P, Demir Y, Gulçin İ (2019) Novel 2-aminopyridine liganded Pd(II) N-heterocyclic carbene complexes: synthesis, characterization, crystal structure and bioactivity properties. Bioorg Chem 91:103134CrossRefGoogle Scholar
  22. Gado AM, Aldahmash BA (2013) Antioxidant effect of Arabic gum against mercuric chloride-induced nephrotoxicity. Drug Des Devel Ther 7:1245CrossRefGoogle Scholar
  23. Gündoğdu S, Türkeş C, Arslan M, Demir Y, Beydemir Ş (2019) New Isoindole-1,3-dione substituted sulfonamides as potent inhibitors of carbonic anhydrase and Acetylcholinesterase: design, synthesis, and biological evaluation. ChemistrySelect 4(45):13347–13355CrossRefGoogle Scholar
  24. Huseynova M, Taslimi P, Medjidov A, Farzaliyev V, Aliyeva M, Gondolova G, Şahin O, Yalçın B, Sujayev A, Orman EB (2018) Synthesis, characterization, crystal structure, electrochemical studies and biological evaluation of metal complexes with thiosemicarbazone of glyoxylic acid. Polyhedron 155:25–33CrossRefGoogle Scholar
  25. Jaishankar M, Tseten T, Anbalagan N, Mathew BB, Beeregowda KN (2014) Toxicity, mechanism and health effects of some heavy metals. Interdiscip Toxicol 7:60–72CrossRefGoogle Scholar
  26. Jarvis P, Quy K, Macadam J, Edwards M, Smith M (2018) Intake of lead (Pb) from tap water of homes with leaded and low lead plumbing systems. ScTEn 644:1346–1356Google Scholar
  27. Kaya ED, Söyüt H, Beydemir Ş (2013) Carbonic anhydrase activity from the gilthead sea bream (Sparus aurata) liver: the toxicological effects of heavy metals. Environ Toxicol Pharmacol 36:514–521CrossRefGoogle Scholar
  28. Khan A, Ahmad L, Khan MZ (2012) Hemato-biochemical changes induced by Pyrethroid insecticides in avian, fish and mammalian species. Int J Agric Biol 14Google Scholar
  29. Kirici M, Demir Y, Beydemir S, Atamanalp M (2016) The effect of Al3 and Hg2 on glucose 6-phosphate dehydrogenase from Capoeta Umbla kidney. Appl Ecol Environ Res 14:253–264CrossRefGoogle Scholar
  30. Kirici M, Turk C, Caglayan C, Kirici M (2017) Toxic effects of copper sulphate pentahydrate on antioxidant enzyme activities and lipid peroxidation of freshwater fish Capoeta umbla (Heckel, 1843) tissues. Appl Ecol Environ Res 15:1685–1696CrossRefGoogle Scholar
  31. Koksal Z, Kalin R, Gulcin I, Ozdemir H (2018) Inhibitory effects of selected pesticides on peroxidases purified by affinity chromatography. Int J Food Prop 21:385–394CrossRefGoogle Scholar
  32. Kose LP, Gülçin İ, Özdemir H, Atasever A, Alwasel SH, Supuran CT (2016) The effects of some avermectins on bovine carbonic anhydrase enzyme. J Enzyme Inhib Med Chem 31:773–778CrossRefGoogle Scholar
  33. Krämer W, Schirmer U, Jeschke P, Witschel M (2012) Modern crop protection compounds, 3 volume set, 1. John Wiley & Sons, HobokenGoogle Scholar
  34. Kucuk M, Gulcin İ (2016) Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity. Environ Toxicol Pharmacol 44:134–139CrossRefGoogle Scholar
  35. Kucukoglu K, Gul HI, Taslimi P, Gulcin I, Supuran CT. (2019) Investigation of inhibitory properties of some hydrazone compounds on hCA I, hCA II and AChE enzymes. Bioorg Chem 86:316–321CrossRefGoogle Scholar
  36. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680CrossRefGoogle Scholar
  37. Manyin T, Rowe CL (2009) Bioenergetic effects of aqueous copper and cadmium on the grass shrimp, Palaemonetes pugio. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 150:65–71Google Scholar
  38. Murty AS (1986) Toxicity of pesticides to fish, 2. CRC press Boca Raton, FL, USAGoogle Scholar
  39. Özaslan MS, Demir Y, Aksoy M, Küfrevioğlu ÖI, Beydemir Ş (2018) Inhibition effects of pesticides on glutathione-S-transferase enzyme activity of Van Lake fish liver. J Biochem Mol Toxicol 32:e22196CrossRefGoogle Scholar
  40. Öztaskın N, Taslimi P, Maraş A, Gülcin İ, Göksu S (2017) Novel antioxidant bromophenols with acetylcholinesterase, butyrylcholinesterase and carbonic anhydrase inhibitory actions. Bioorg Chem 74:104–114CrossRefGoogle Scholar
  41. Öztaşkın N, Kaya R, Maraş A, Şahin E, Gülcin İ, Göksu S (2019) Synthesis and characterization of novel Bromophenols: determination of their anticholinergic, Antidiabetic and antioxidant activities. Bioorg ChemGoogle Scholar
  42. Pari L, Karthikeyan A, Karthika P, Rathinam A (2015) Protective effects of hesperidin on oxidative stress, dyslipidaemia and histological changes in iron-induced hepatic and renal toxicity in rats. Toxicol Rep 2:46–55CrossRefGoogle Scholar
  43. Peterson RE, Tu C, Linser PJ (1997) Isolation and characterization of a carbonic anhydrase homologue from the zebrafish (Danio rerio). JMolE 44:432–439Google Scholar
  44. Rai PK (2009) Heavy metal phytoremediation from aquatic ecosystems with special reference to macrophytes. Crit Rev Environ Sci Technol 39:697–753CrossRefGoogle Scholar
  45. Sağlık BN, Çevik UA, Osmaniye D, Levent S, Çavuşoğlu BK, Demir Y, Ilgın S, Özkay Y, Koparal AS, Beydemir Ş, Kaplancıklı ZA (2019) Synthesis, molecular docking analysis and carbonic anhydrase I-II inhibitory evaluation of new sulfonamide derivatives. Bioorg Chem 91:103153CrossRefGoogle Scholar
  46. Sender S, Böttcher K, Cetin Y, Gros G (1999) Carbonic anhydrase in the gills of seawater-and freshwater-acclimated flounders Platichthys flesus: purification, characterization, and immunohistochemical localization. J Histochem Cytochem 47:43–50CrossRefGoogle Scholar
  47. Simonsen LO, Harbak H, Bennekou P (2012) Cobalt metabolism and toxicology—a brief update. ScTEn 432:210–215Google Scholar
  48. Sivaperumal P, Sankar T, Nair PV (2007) Heavy metal concentrations in fish, shellfish and fish products from internal markets of India Vis-a-Vis international standards. Food Chem 102:612–620CrossRefGoogle Scholar
  49. Soyut H, Beydemir S (2008) Purification and some kinetic properties of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) liver and metal inhibition. Protein and peptide letters 15:528–535CrossRefGoogle Scholar
  50. Söyüt H, Beydemir Ş (2012) The impact of heavy metals on the activity of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) kidney. Toxicol Ind Health 28:296–305CrossRefGoogle Scholar
  51. Söyüt H, Beydemir Ş, Ceyhun SB, Erdoğan O, Kaya ED (2012) Changes in carbonic anhydrase activity and gene expression of Hsp70 in rainbow trout (Oncorhynchus mykiss) muscle after exposure to some metals. Turk J Vet Anim Sci 36:499–508Google Scholar
  52. Taslimi P, Caglayan C, Gulcin İ (2017) The impact of some natural phenolic compounds on carbonic anhydrase, acetylcholinesterase, butyrylcholinesterase, and α-glycosidase enzymes: an antidiabetic, anticholinergic, and antiepileptic study. J Biochem Mol Toxicol 31:e21995CrossRefGoogle Scholar
  53. Taslimi P, Caglayan C, Farzaliyev V, Nabiyev O, Sujayev A, Turkan F, Kaya R, Gulçin İ (2018a) Synthesis and discovery of potent carbonic anhydrase, acetylcholinesterase, butyrylcholinesterase, and α-glycosidase enzymes inhibitors: the novel N, N′-bis-cyanomethylamine and alkoxymethylamine derivatives. J Biochem Mol Toxicol 32:e22042CrossRefGoogle Scholar
  54. Taslimi P, Osmanova S, Caglayan C, Turkan F, Sardarova S, Farzaliyev V, Sujayev A, Sadeghian N, Gulçin İ (2018b) Novel amides of 1, 1-bis-(carboxymethylthio)-1-arylethanes: synthesis, characterization, acetylcholinesterase, butyrylcholinesterase, and carbonic anhydrase inhibitory properties. J Biochem Mol Toxicol 32:e22191CrossRefGoogle Scholar
  55. Taslimi P, Kandemir FM, Demir Y, İleritürk M, Temel Y, Caglayan C, Gulçin İ (2019) The antidiabetic and anticholinergic effects of chrysin on cyclophosphamide-induced multiple organ toxicity in rats: Pharmacological evaluation of some metabolic enzyme activities. J Biochem Mol Toxicol:e22313Google Scholar
  56. Tchounwou PB, Yedjou CG, Patlolla AK, Sutton DJ (2012) Heavy metal toxicity and the environment, molecular, clinical and environmental toxicology. Springer, Berlin, pp 133–164CrossRefGoogle Scholar
  57. Topal M, Gülçin İ (2014) Rosmarinic acid: a potent carbonic anhydrase isoenzymes inhibitor. Turk J Chem 38:894–902CrossRefGoogle Scholar
  58. Turk E, Kandemir FM, Yildirim S, Caglayan C, Kucukler S, Kuzu M (2019) Protective effect of hesperidin on sodium arsenite-induced nephrotoxicity and hepatotoxicity in rats. Biol Trace Elem Res 189:95–108CrossRefGoogle Scholar
  59. Turkan F, Cetin A, Taslimi P, Karaman M, Gulçin İ (2019) Synthesis, biological evaluation and molecular docking of novel pyrazole derivatives as potent carbonic anhydrase and acetylcholinesterase inhibitors. Bioorg Chem 86:420–427CrossRefGoogle Scholar
  60. Türkeş C, Arslan M, Demir Y, Çoçaj L, Rifati Nixha A, Beydemir Ş (2019) Synthesis, biological evaluation and in silico studies of novel N-substituted phthalazine sulfonamide compounds as potent carbonic anhydrase and acetylcholinesterase inhibitors. Bioorg Chem 89:103004CrossRefGoogle Scholar
  61. Vakonaki E, Androutsopoulos VP, Liesivuori J, Tsatsakis AM, Spandidos DA (2013) Pesticides and oncogenic modulation. Toxicology 307:42–45CrossRefGoogle Scholar
  62. Walia S, Sharma RK, Parmar BS (2009) Isolation and simultaneous LC analysis of thiram and its less toxic transformation product in DS formulation. Bull Environ Contam Toxicol 83:363–368CrossRefGoogle Scholar
  63. Wilbur KM, Anderson NG (1948) Electrometric and colorimetric determination of carbonic anhydrase. J Biol Chem 176:147–154Google Scholar
  64. Ye C, Zhou Q, Wang X (2008) Determination of thiophanate-methyl and chlorotoluron in water samples by improved single-drop microextraction coupled with high-performance liquid chromatography. Int J Environ Anal Chem 88:461–471CrossRefGoogle Scholar
  65. Yousef MI, Awad TI, Mohamed EH (2006) Deltamethrin-induced oxidative damage and biochemical alterations in rat and its attenuation by vitamin E. Toxicology 227:240–247CrossRefGoogle Scholar
  66. Zengin M, Genc H, Taslimi P, Kestane A, Guclu E, Ogutlu A, Karabay O, Gulçin İ (2018) Novel thymol bearing oxypropanolamine derivatives as potent some metabolic enzyme inhibitors–their antidiabetic, anticholinergic and antibacterial potentials. Bioorg Chem 81:119–126CrossRefGoogle Scholar

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Authors and Affiliations

  1. 1.Department of Biochemistry, Faculty of Veterinary MedicineBingol UniversityBingolTurkey
  2. 2.Department of Biotechnology, Faculty of ScienceBartin UniversityBartinTurkey
  3. 3.Department of Fisheries, Genç Vocational SchoolBingol UniversityBingolTurkey
  4. 4.Department of Chemistry, Faculty of ScienceAtatürk UniversityErzurumTurkey
  5. 5.Department of Biochemistry, Faculty of Veterinary MedicineAtatürk UniversityErzurumTurkey
  6. 6.Department of Pharmacy Services, Nihat Delibalta Göle Vocational High SchoolArdahan UniversityArdahanTurkey
  7. 7.Department of Biochemistry, Faculty of PharmacyAnadolu UniversityEskişehirTurkey

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