ISOLATION OF ANGIOTENSIN-CONVERTING ENZYME INHIBITOR PRODUCING BACTERIA FROM COW MILK
Objective: To evaluate the potential of protease producing organism for the production of Angiotensin Iâ€“converting enzyme (ACE) inhibitor by fermentation of various protein substrates.
Methods: Bacterial strains were isolated from cow milk collected in Coimbatore, Tamil Nadu, India by using serial dilution technique, plated on nutrient agar medium. The identity of the strain was ascertained by 16s rRNA gene sequencing method and was submitted to the NCBI GenBank nucleotide database. Various substrates were screened for ACE inhibitor production by the fermentation with the isolated strain.
Results: The isolated coded as BUCTL09, which showed a significant zone of clearance was selected and identified as Micrococcus luteus (KF303592.1). Among the seven substrates, only beef extract fermented broth showed an inhibition of 79% and was reported as the best substrate.
Conclusion: In the search for non-toxic, and economic ACE inhibitors as an alternative to the synthetic drugs, many natural ACE inhibitors have been isolated from a microbial source. In the present study, isolate BUCTL09 was selected for the production of ACE inhibitor from the beef extract. Findings from this study lead us to investigate this potent ACE inhibitor further for its biological properties and to explore the impending efficacy of the ACE inhibitor which may conceivably be developed into a prospective drug.
2. Takano Y. Object shift and scrambling. Natural Language Linguistic Theory 1998;16:817-89.
3. Deshayes S, Nahmias O. Angiotensin receptors: a new role in cancer? Trends Endocrinol Metab 2005;16:1016â€“8.
4. Galil AOA. Formation of proteases by Aspergillus fumigatus and Pencillium sp. J King Saud Univ Sci 1992;4:127-36.
5. Kandler O, Weiss N. Genus Lactobacillus Beijerinck 1901. 212AL. In: Sneath PHA, Mair NS, Sharpe ME, Holt JG. editors. Bergeyâ€™s manual of systematic bacteriology. Baltimore: Williams and Wilkins Co; 1986. p. 1209â€“34.
6. Sambrook J, Fritsch FE, Maniatis T. Molecular cloning: a laboratory manual. 2nd. ed. New York: Cold Spring Harbour; 1989.
7. Altschul S, Madden T, Schaffer A. Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 1997;25:3389â€“402.
8. Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties, and weight matrix choice. Nucleic Acids Res 1994;22:4673â€“80.
9. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951;193:265-75.
10. Cushman DW, Cheung HS. Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem Pharmacol 1971;20:1637-48.
11. Garbutt. Essentials of food microbiology. Arnold International edition; 1997. p. 67-72.
12. Sharpe ME, Fryer TF, Smith DG. Identification of lactic acid bacteria. In: Gibbs EM, Skinner FA. editors. Identification methods for microbiologists. London: Academic Press; 1979. p. 233-59.
13. Cote RJ. Media composition, microbial, laboratory scale. In: Flickinger MC, Drew SW. editors. Encyclopedia of bioprocess technology: Fermentation, biocatalysis, and bioseparation. New York: John Wiley and Sons Inc; 1999;1-5:1640-60.
14. Bridson EY, Brecker A. Design and formulation of microbial culture media. Methods Microbiol 1970;3:229-95.
15. Lindberg H, Nielsen D, Jensen BV, Eriksen J, Skovsgaard T. Angiotensin converting enzyme inhibitors for cancer treatment? Acta Oncol 2004;43:142â€“52.
16. Lever A, Hole DJ, Gillis CR, McCallum IR, McInnes GT, MacKinnon PL, et al. Do inhibitors of angiotensin-I-converting enzyme protect against risk of cancer? Lancet 1998;352:179â€“84.