MICROBICIDAL POTENTIALITY OF PURIFIED ANTHOCYANIN FROM IN VITRO CULTURE OF CLERODENDRON INFORTUNATUM L. AGAINST SELECTED PATHOGENS

Meenu Krishnan V. G.; Greeshma Murukan; Aswathy J. M.; Bosco Lawarence; Murugan K.

doi:10.22159/ijpps.2018v10i6.18649

Authors

Meenu Krishnan V. G. Plant Biochemistry and Molecular Biology Laboratory, Department of Botany, University College, Trivandrum, 695034, Kerala
Greeshma Murukan Plant Biochemistry and Molecular Biology Laboratory, Department of Botany, University College, Trivandrum, 695034, Kerala
Aswathy J. M. Plant Biochemistry and Molecular Biology Laboratory, Department of Botany, University College, Trivandrum, 695034, Kerala
Bosco Lawarence Plant Biochemistry and Molecular Biology Laboratory, Department of Botany, University College, Trivandrum, 695034, Kerala
Murugan K. Plant Biochemistry and Molecular Biology Laboratory, Department of Botany, University College, Trivandrum, 695034, Kerala

DOI:

https://doi.org/10.22159/ijpps.2018v10i6.18649

Keywords:

Clerodendron, HPLC, MS medium, Hormones, Anthocyanin, Column chromatography, Antimicrobial

Abstract

Objective: Clerodendron infortunatum L. is a widely used medicinal herb over centuries for curing many skin-borne disorders. The present study was designed to validate the tribal knowledge by evaluating antimicrobial potential of purified anthocyanin extracted from in vitro cell suspension culture.

Methods: The explants were inoculated on murashige and skoog (MS) medium mixed with various combinations of 2, 4-D a+BAP for callus induction. Green compact callus was initiated within 30 d from the explants on MS medium fortified with benzylaminopurine (BAP) (2.0 mg/l)+2, 4-D (0.5 mg/l). Subsequently, anthocyanin was triggered from the compact callus by subculturing in the medium containing 2, 4-D and Kinetin. Cell suspension culture was also developed. Anthocyanin production was enhanced by elicitation using salicylic acid and others. Three chromatographic methods such as solid phase extraction by Sepharose C18 column, Oasis-MCX and Amberlite XAD 7+Sephadex LH 120 sorbents were used to purify the in vitro synthesized anthocyanin from the cell cultures. HPLC and molar absorptivity assay were carried to check the purity. Antimicrobial analysis was also carried using standard protocols to check minimum inhibitory concentration (MIC) and minimum killing concentration (MKC).

Results: The mean purity values obtained by high-performance thin layer chromatography (HPLC) were 90.9%Â±1.9, 80.60%Â±2.3 for Oasis MCX, Amberlite XAD-7+Sephadex LH-20 column respectively. However, the purity by molar absorptivity was found to be less. HPLC chromatogram revealed 12 fractions of anthocyanin. Inhibition zone diameter, MIC and MKC values obtained for the purified anthocyanin revealedÂ its antimicrobial potentiality but at different levels among the selected bacteria and fungi. C.Â albicans, S. aureus, P. aerugenosa showed significant values followed by MRSA, E. coli and A. flavus. The results are comparable with the synthetic antibiotics. However, E. faecalis was more resistance. Mode of action was confirmed from the results of intracellular potassium leakage and bacterial membrane integrity analysis.

Conclusion: Thus, the study confirms the efficacy of anthocyanin as natural antimicrobial and suggests the possibility of employing it as drugs for the treatment of infectious diseases caused by the pathogens.

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