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Biological Properties and Characterization of ASL50 Protein from Aged Allium sativum Bulbs

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Abstract

Allium sativum is well known for its medicinal properties. The A. sativum lectin 50 (ASL50, 50 kDa) was isolated from aged A. sativum bulbs and purified by gel filtration chromatography on Sephacryl S-200 column. Agar well diffusion assay were used to evaluate the antimicrobial activity of ASL50 against Candida species and bacteria then minimal inhibitory concentration (MIC) was determined. The lipid A binding to ASL50 was determined by surface plasmon resonance (SPR) technology with varying concentrations. Electron microscopic studies were done to see the mode of action of ASL50 on microbes. It exerted antimicrobial activity against clinical Candida isolates with a MIC of 10–40 μg/ml and clinical Pseudomonas aeruginosa isolates with a MIC of 10–80 μg/ml. The electron microscopic study illustrates that it disrupts the cell membrane of the bacteria and cell wall of fungi. It exhibited antiproliferative activity on oral carcinoma KB cells with an IC50 of 36 μg/ml after treatment for 48 h and induces the apoptosis of cancer cells by inducing 2.5-fold higher caspase enzyme activity than untreated cells. However, it has no cytotoxic effects towards HEK 293 cells as well as human erythrocytes even at higher concentration of ASL50. Biological properties of ASL50 may have its therapeutic significance in aiding infection and cancer treatments.

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Abbreviations

LA:

Lipid A

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

MRSA:

Methicillin-resistant Staphylococcus aureus

MHA:

Mueller Hinton agar

SDA:

Sabouraud dextrose agar

MIC:

Minimal inhibitory concentration

CLSI:

Clinical and Laboratory Standards Institute

MHB:

Mueller Hinton broth media

TEM:

Transmission electron microscopy

SEM:

Scanning electron microscopy

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Acknowledgments

The authors acknowledge the Indian Council of Medical Research, Government of India, New Delhi, India, for providing funds for the consumable items and Electron Microscopic Facility, AIIMS, for EM studies.

Ethics

The Ethics Committee of All India Institute of Medical Sciences, (AIIMS) New Delhi, India, approved the study protocol (IEC/NP-374/2013) and informed consent was obtained.

Conflict of Interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Department of Biophysics, All India Institute of Medical Sciences, New Delhi, 110029, India

    Suresh Kumar, Kumar Jitendra, Kusum Singh, Mou Sinha, Sujata Sharma, Tej P. Singh & Sharmistha Dey

  2. Department of Biotechnology, All India Institute of Medical Sciences, New Delhi, 110029, India

    Vaishali Kapoor & Satya N. Das

  3. Department of Microbiology, All India Institute of Medical Sciences, New Delhi, 110029, India

    Immaculata Xess

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  1. Suresh Kumar
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  2. Kumar Jitendra
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  3. Kusum Singh
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Correspondence to Sharmistha Dey.

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Kumar, S., Jitendra, K., Singh, K. et al. Biological Properties and Characterization of ASL50 Protein from Aged Allium sativum Bulbs. Appl Biochem Biotechnol 176, 1914–1927 (2015). https://doi.org/10.1007/s12010-015-1687-y

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  • DOI: https://doi.org/10.1007/s12010-015-1687-y

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