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American Journal of Drug Discovery and Development

Year: 2013 | Volume: 3 | Issue: 3 | Page No.: 120-129
Research Article

Isolation, Partial Purification and Characterization of Angiotensin Converting Enzyme (ACE) from Rabbit (Oryctolagus ciniculus) Lungs

A. Abdulazeez Mansurah
Department of Biochemistry, Faculty of Science, Ahmadu Bello University, Zaria, Nigeria
LiveDNA: 234.30336
I.A. Aimola
Department of Developmental and Regenerative Biology, Mount Sinai School of Medicine, New York
Rukevwe O. Annette
Department of Biochemistry, Faculty of Science, Ahmadu Bello University, Zaria, Nigeria
S. Abdullahi
Department of Biochemistry, Faculty of Science, Ahmadu Bello University, Zaria, Nigeria
A simple method was set up to isolate, partially purify and characterize angiotensin-converting enzyme from rabbit lungs. Angiotensin-converting enzyme was purified by a three-step method: Ammonium sulphate precipitation (30-70%), gel filtration chromatography (Sephadex G-75) and ion exchange chromatography (CM-sephadex). The enzyme activity was assayed by monitoring the rate of production of hippuric acid from the hydrolysis of Hippuryl-L-Histidine-L-Leucine by angiotensin-converting enzyme. From the results, the enzyme was purified 6.25-fold with a yield of 21% on CM-Sephadex column. The angiotensin converting enzyme from rabbit lung had a broad optimum pH range (8.0-8.3) and optimum temperature of 37°C. Initial velocity studies for the determination of kinetic constants with Hippuryl-L-Histidine-L-Leucine as substrate revealed a KM and VMAX of 1.8 and 0.4 μmol min-1, respectively. The enzyme activity was not affected by Mg2+, Ca2+, Na+ and K+, while EDTA strongly inhibited it. In conclusion, this study has shown that though angiotensin-converting enzyme can be partially-purified from rabbit lungs via. a three-step purification protocol, the purification steps and assay conditions needs to be modified to obtain a higher yield and specific activity.
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How to cite this article

A. Abdulazeez Mansurah, I.A. Aimola, Rukevwe O. Annette and S. Abdullahi, 2013. Isolation, Partial Purification and Characterization of Angiotensin Converting Enzyme (ACE) from Rabbit (Oryctolagus ciniculus) Lungs. American Journal of Drug Discovery and Development, 3: 120-129.

DOI: 10.3923/ajdd.2013.120.129

URL: https://scialert.net/abstract/?doi=ajdd.2013.120.129

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