Optimization, Chromatography-based Purification and characterization of novel Antibacterial compound 4-formyl-2-hydroxy bicyclo [4.1.0] heptane-7-carboxylic acid produced by Streptomyces rochei (OM746935)

Author: Dhanasekaran Kokila, Venkatapathi Bhuvaneswari, Manickam Ramesh, Natarajan Ranjani and Ranganathan Kannan

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Abstract

Actinobacteria are the most affordable source of novel antibiotic production for the biotechnological and pharmaceutical industries. In the present work, Streptomyces rochei was isolated from Bargur hills and was investigated for their antimicrobial activity by agar well diffusion method, Streptomyces rochei was active against all the test organisms, and the maximum zone of inhibition was observed in Staphylococcus aureus lowest concentration of MIC 1.25 mg/ml. Ethyl acetate extract showed four fractions in TLC and the active fraction was identified by bioautography. This study investigates the optimization of the nutrient content of the fermentation medium by alteration in carbon and nitrogen source, medium PH, and incubation days are monitored. Then purification of bioactive compounds by column chromatography, 30 fractions were obtained 21, 23, and 24th fractions active against all the tested organisms. All these active fractions are again purified by preparative HPLC which contains C-18 column. Sixteen fractions were again investigated for antibacterial activity, the best activity was obtained in the 7th fraction. Significant peaks were visible in the HPLC chromatogram at the 11th and 16th minutes. Using a UV detector, the maximum absorbance was recorded in the UV range of 220 nm. The active fraction was further characterized by FTIR, GCMS, and NMR. Based on the NMR shift values structure and name of the compound are predicted as 4-formyl-2-hydroxy bicyclo [4.1.0] heptane-7-carboxylic acid. This compound is responsible for the action of antibacterial activity. Actinomycetes isolated from terrestrial sources can each produce a variety of secondary metabolites; in fact, the vast majority of antibiotics originate from these sources. However, a number of diseases are developing resistance to the widely prescribed antibiotics. The development of drugs to stop the spread of these infections is therefore urgently needed.

Keywords

Optimization, column chromatography, preparative HPLC, FTIR, GC-MS

Conclusion

Strain Streptomyces rochei BF3A has the ability to produce antibacterial compounds. Purified antibacterial compound structurally identified as 4-formyl-2-hydroxy bicyclo [4.1.0] heptane-7-carboxylic acid. This compound is active against all the test organisms particularly the maximum zone of inhibition in Staphylococcus aureus. The Insilco docking approaches of a compound with the target sites of Staphylococcus aureus will be studied in the future.

References

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How to cite this article

Dhanasekaran Kokila, Venkatapathi Bhuvaneswari, Manickam Ramesh, Natarajan Ranjani and Ranganathan Kannan (2023). Optimization, Chromatography-based Purification and characterization of novel Antibacterial compound 4-formyl-2-hydroxy bicyclo [4.1.0] heptane-7-carboxylic acid produced by Streptomyces rochei(OM746935). Biological Forum – An International Journal, 15(5): 1352-1361.