Research article
Volume 13 Issue 6 - 2022
Purification and characterization of a sialic Acid specific bacterial lectin from Corynebacterium amycolatum and assigning its role in Keratitis

Narasimhappagari Jagadeesh1, Shivakumar Belur1, Sanhita Roy2 and Shashikala R Inamdar1*

1Department of Studies in Biochemistry, Karnatak University, Dharwad, India
2Professor Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, India
*Corresponding Author: Shashikala R Inamdar, Department of Studies in Biochemistry, Karnatak University, Dharwad, India.
Received: April 02, 2022; Published: May 25, 2022




Abstract

A sialic acid-specific bacterial lectin, CAL from Corynebacterium amycolatum, which was isolated from the corneal smears of a keratitis patient was purified by 78-fold with a combination of anion exchange chromatography using DEAE-cellulose and affinity chromatography on fetuin coupled Sepharose 4B column with an overall recovery of 45%. The purified lectin is a monomer and showed apparent molecular mass of 19.7 kDa as elucidated by SDS-PAGE both under reducing and non reducing conditions. CAL is human blood group non specific lectin which agglutinates trypsinised and neuraminidase treated human and rabbit erythrocytes. CAL is an N-acetyl neuraminic acid (NANA) specific lectin with least MIC but it also recognizes Fetuin, Mucin, Glucose, Galactose and Lactose. CAL showed strong binding to HCECs which could be blocked by NANA and mucin. CAL showed mitogenic effect on HCECs at concentrations below 0.3 µg/mL leading to expression of proinflammatory cytokines IL-6 and IL-8. In contrast, CAL showed growth inhibitory effects on HCECs above 0.6 µg/mL concentrations at 48h with an IC50 of 1.2 µg/mL. Annexin- V/PI assay results shows that, at higher concentration, CAL induced apoptotic effect in HCECs with an increase in early and late apoptotic population. These findings demonstrate that CAL has a significant role in mediating host pathogen interactions eventually leading to pathogenesis and hence are of clinical significance in developing strategy in controlling the infection or bacterial keratitis.

Keywords: Corynebacterium amycolatum Lectin; Bacterial Keratitis; Sialic Acid Specific; Proinflammatory; Apoptotisis

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Citation: Shashikala R Inamdar., et al. “Purification and characterization of a sialic Acid specific bacterial lectin from Corynebacterium amycolatum and assigning its role in Keratitis”. EC Ophthalmology 13.5 (2022): 03-17.

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