Investigation of inhibition effect of folic acid (vitamin B9) on angiotensin-converting enzyme activity purified from human plasma

Abstract

Inhibition of the angiotensin-converting enzyme (ACE, EC 3.4.15.1) is one of the most important hypertension treatments. Here, ACE was purified from human plasma with affinity chromatography. The purity and molecule weight of ACE were identified utilizing the SDS-PAGE and viewed in two bands at around 60 kDa and 70 kDa on the gel. KM and Vmax constants from the Lineweaver-Burk graphic were computed to be 0.6 mM and 175.44 (µmol/min).mL-1 , respectively. The effects of folic acid (vitamin B9) on purified ACE were studied. Folic acid on purified ACE demonstrated an inhibitory efficacy. The IC50 value for folic acid was calculated to be 127.94 µM. Kind of inhibitory and Ki constant for folic acid were defined. The kind of inhibitory for folic acid was found as non-competitive inhibitory. Ki constant was computed to be 226.59 µM for folic acid. In this study, it was concluded that folic acid, which shows an inhibitor efficacy on ACE, may have both therapeutic and protective impacts against hypertension.

Keywords

• angiotensin-converting enzyme
• folic acid (vitamin B9)
• inhibition
• purification

References

1. Andújar-Sánchez M, Cámara-Artigas A, Jara-Pérez V, 2003. Purification of angiotensin I converting enzyme from pig lung using concanavalin-A sepharose chromatography. Journal of Chromatography B, 783: 247-252.
2. Aydin F, Turkoglu V, Bas Z, 2021. Purification and characterization of angiotensin-converting enzyme (ACE) from sheep lung, Molecular Biology Reports 48: 4191-4199.
3. Bas Z, 2021. Inhibition effect of nicotinamide (vitamin B3) and reduced glutathione (GSH) peptide on angiotensin-converting enzyme activity purified from sheep kidney, International Journal of Biological Macromolecules, 189: 65-71.
4. Bas Z, Turkoglu V, Goz Y, 2022. Investigation of inhibition effect of butanol and water extracts of Matricaria chamomilla L. on angiotensin-converting enzyme purified from human plasma. Biotechnology and Applied Biochemistry, 69: 273–280.
5. Basi Z, Turkoglu V, 2018. Purification of angiotensin-converting enzyme from human plasma and investigation of the effect of some active ingredients isolated from Nigella sativa L.extract on the enzyme activity. Biomedical Chromatography, 32: e4175.
6. Basi Z, Turkoglu V, 2019. In vitro effect of oxidized and reduced glutathione peptides on angiotensin converting enzyme purified from human plasma. Journal of Chromatography B Analyt. Technol. Biomed. Life Sci., 1104: 190-195.
7. Basi Z, Turkoglu N, Turkoglu V, Karahan F, 2019. In vitro effect of ethyl acetate, butanol and water extracts of Juniperus excelsa Bieb. on angiotensin-converting enzyme purified from human plasma. Chemical Papers, 73: 2525–2533.
8. Bernstein KE, Khan Z, Giani, JF, Cao DY, Bernstein, EA, Shen XZ, 2018. Angiotensin-converting enzyme in innate and adaptive immunity. Nature reviews. Nephrology,14: 325–336.

9. Botez MI, 1976. Folate deficiency and neurological disorders in adults. Med Hypotheses. 1976; 2(4):135-40.
10. Bradford MM, 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-54.
11. Choi JH, Yates Z, Veysey M, Heo YR, Lucock M, 2014. Contemporary issues surrounding folic Acid fortification initiatives. Preventive Nutrition and Food Science, 19 (4): 247–260.
12. Çelikezen FÇ, Türkoğlu V, Fırat M, Baş Z, 2021. The Effects of Coriandrum sativum L. and Chaerophyllum macropodum Boiss. (Apiaceae) on human plasma angiotensin-converting enzyme (ACE) in vitro. BEU Journal of Science, 10 (3): 710-718.
13. Fu W, Chen C, Zeng H, Lin J, Zhang Y, Hu J, Zheng B, 2019. Novel angiotensin-converting enzyme inhibitory peptides derived from Trichiurus lepturus myosin: Molecular docking and surface plasmon resonance study. LWT, 110: 54-63.
14. Gür T, Karahan F, Baş Z, Türkoğlu V, 2020. The Determination of Inhibition Effect of Extracts of Thymbra sintenisii Bornm. et Aznav. Subsp on Angiotensin Converting Enzyme. Journal of the Institute of Science and Technology, 10 (3): 1848-1856.
15. Holmquist B, Bünning P, Riordan JF, 1979. A continuous spectrophotometric assay for angiotensin converting enzyme. Analytical Biochemistry, 95 (2): 540-548.
16. Huo Y, Qin X, Wang J, Sun N, Zeng Q, Xu X, Liu, L Xu, X, Wang X, 2012. Efficacy of folic acid sup¬plementation in stroke prevention: new insight from a meta-analysis. International Journal of Clinical Practice, 66 (6): 544-51.
17. Karahan F, Turkoglu V, 2021 Enhanced purification protocol for the angiotensin-converting enzyme from bovine systems and investigation of the in vitro effect of some active substances Chemico-Biological Interactions, 347: 109604.
18. Kiylik A, Turkoglu V, Bas Z, 2022. Purification of angiotensin-converting enzyme (ACE) from sheep kidney and inhibition effect of reduced nicotinamide adenine dinucleotide (NADH) on purified ACE activity. Cell Biochemistry and Biophysics, 80: 115-122.
19. Laemmli UK, 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685.
20. Li J, Liu, Z, Zhao Y, Zhu X, Yu R, Dong S, Wu H, 2018. Novel natural angiotensin converting enzyme (ACE)-inhibitory peptides derived from sea cucumber-modified hydrolysates by adding exogenous proline and a study of their structure–activity relationship. Marine Drugs, 16 (8): 271.
21. Liew SC, 2016. Folic acid and diseases - Supplement it or not? Revista da Associação Médica Brasileira, 62 (1): 90-100.
22. Lineweaver H, Burk D, 1934. The determination of enzyme dissociation constants. Journal of the American Chemical Society, 56: 658–660.
23. Liu CL, Fang L, Min WH, Liu JS, Li HM, 2018. Exploration of the molecular interactions between angiotensin-I-converting enzyme (ACE) and the inhibitory peptides derived from hazelnut (Corylus heterophylla Fisch.). Food Chemistry, 245: 471–480.
24. McRae MP, 2009. High-dose folic acid supplementation effects on endothelial function and blood pressure in hypertensive patients: a meta-analysis of randomized controlled clinical trials. Journal of Chiropractic Medicine, 8 (1): 15-24.
25. Pantoliano MW, Holmquist B, Riordan JF, 1984. Affinity chromatographic purification of angiotensin converting enzyme. Biochemistry, 23 (5): 1037-1042.
26. Qin X, Xu M, Zhang, Y, Li J, Xu X, Wang X, Xu X, Huo Y, 2012. Effect of folic acid supplementation on the progression of carotid intima-media thickness: a meta-analysis of randomized controlled trials. Atherosclerosis, 222 (2): 307-313.
27. Sabeur K, Vo AT, Ball BA, 2001. Characterization of angiotensin-converting enzyme in canine testis. Reproduction, 122 (1): 139-146.
28. Strittmatter SM, Thiele EA, Kapiloff MS, Snyder SH, 1985. A rat brain isozyme of angiotensin-converting enzyme unique specificity for amidated peptide substrates. The Journal of Biological Chemistry, 260 (17): 9825-9832. Tu M, Wang C, Chen C, Zhang R, Liu H, Lu W, Jiang L, Du M, 2018. Identification of a novel ACE-inhibitory peptide from casein and evaluation of the inhibitory mechanisms. Food Chemistry, 256: 98-104.
29. Weinstein SJ, Hartman TJ, Stolzenberg-Solomon R, Pietinen P, Barrett MJ, Taylor PR, Virtamo, J, Albanes D, 2003. Null association between prostate cancer and serum folate, vitamin B(6), vitamin B(12), and homocysteine. Cancer Epidemiology Biomarkers  Prevention, 12: 1271-1272.
30. West AA, Caudill MA, Bailey LB, 2020. "Folate". In BP Marriott, DF Birt, VA Stallings, AA Yates (eds.). Present Knowledge in Nutrition, Eleventh Edition. London, United Kingdom: Academic Press (Elsevier). Pp: 273–88.
31. Yu ZP, Guo H, Shiuan D, Xia CS, Zhao WZ, Ding L, Zheng FP, Liu JB, 2020. Interaction mechanism of egg white-derived ACE inhibitory peptide TNGIIR with ACE and its effect on the expression of ACE and AT1 receptor. Food Science and Human Wellness, 9 (1): 52–57.