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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 volume 80, pages 115–122 (2022)Cite this article

Abstract

Angiotensin-converting enzyme (ACE, EC 3.4.15.1) is a significant enzyme that regulates blood pressure. ACE inhibitors are often used in the treatment of hypertension. In this work, ACE was purified and characterized in one step with affinity chromatography from sheep kidneys. ACE was 10305-fold purified and specific activity was 19,075 EU/mg protein. The molecular weight and purity of ACE were found with SDS-PAGE and observed two bands at about 60 kDa and 70 kDa on the gel. The effects of reduced nicotinamide adenine dinucleotide (NADH), an antioxidant compound, on purified ACE activity were also researched. NADH on ACE activity showed an inhibition effect. The inhibition type of NADH was determined to be non-competitive inhibition by the Lineweaver–Burk chart and IC50 and Ki values for NADH were 244.33 and 175.08 µM, respectively. These results suggest that antioxidant substances might be efficient in preventing hypertension.

Highlights

  • ACE was purified from sheep kidneys.

  • Ki and IC50 for NADH were 175.08 and 244.33 µM, respectively.

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Acknowledgements

This work received financial support from the Head of Scientific Research Projects of Van Yüzüncü Yıl University (FYL-2018-7037).

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Affiliations

  1. Department of Chemistry, Faculty of Science, Van YüzüncüYıl University, Van, Turkey

    Aysenur Kiylik & Vedat Turkoglu

  2. Department of Nutrition and Dietetics, Faculty of Health Sciences, Van Yüzüncü Yıl University, Van, Turkey

    Zehra Bas

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  1. Aysenur Kiylik
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  2. Vedat Turkoglu
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  3. Zehra Bas
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Contributions

Vedat Turkoglu contributed to the study conception and design. All authors contributed to the manuscript. Vedat Turkoglu, Aysenur Kiylik, and Zehra Bas performed the experiments and analyzed the data. Zehra Bas wrote the first draft of the manuscript. Vedat Turkoglu and Zehra Bas contributed to the revisions and the final draft of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Zehra Bas.

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This work involved the chemical analyses of animal sources. No approval of animal use protocols was required.

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Kiylik, A., Turkoglu, V. & Bas, Z. Purification of Angiotensin-Converting Enzyme (ACE) from Sheep Kidney and Inhibition Effect of Reduced Nicotinamide Adenine Dinucleotide (NADH) on Purified ACE Activity. Cell Biochem Biophys 80, 115–122 (2022). https://doi.org/10.1007/s12013-021-01036-2

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  • DOI: https://doi.org/10.1007/s12013-021-01036-2

Keywords

  • Angiotensin-converting enzyme (ACE)
  • Reduced nicotinamide adenine dinucleotide (NADH)
  • Antioxidant
  • Purification
  • Inhibition