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Kerala Veterinary and Animal Sciences University, Wayanad

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20171
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Subject: Biochemistry × Author: Sinchu Vijayan ×
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    ThesisItemOpen Access
    ISOLATION AND CHARACTERIZATION OF LACTOFERRIN FROM COLOSTRUM OF GOATS
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2017) Sinchu Vijayan; UMA, R.
    Lactoferrin, an 80 k Da iron-binding protein, primarily present in milk, is well known for its multifunctional properties such as antibacterial, antifungal, antiviral, antioxidant, immunomodulatory and anticancer activities. The present study focussed on the isolation and characterization of lactoferrin from the colostrum of Malabari, Attappady Black and crossbred goats of Kerala as well as assessment of the antimicrobial potential of the lactoferrin isolated. Colostrum samples collected from the three goat breeds maintained at University Goat and Sheep Farm, College of Veterinary and Animal Sciences, Mannuthy were processed to remove fat globules and casein. The whey obtained after processing was fractionated with ammonium sulphate to remove globulins from the sample. Fraction containing albumin and remaining proteins including lactoferrin was separated out, dialysed against equilibration buffer, loaded on to CM- Sephadex C-50 cation exchanger column and eluted with a step gradient of 0.4, 0.6 and 0.8 M NaCl. The fractions with high OD280 values were analysed using 12 per cent SDS-PAGE to identify their protein components in comparison with standard protein. The protein fractions with high absorbance at 280nm, eluted with 0.6M NaCl, could be visualised as a single 80 kDa Coomassie Brilliant Blue-stained band. The total iron content in the isolated lactoferrin samples was estimated by atomic absorption spectrophotometry and it was found to be 820 ppm for Malabari and Attappady Black lactoferrin whereas 1100 ppm for crossbred goat lactoferrin. The concentration of Malabari lactoferrin (mgLf), Attappady Black lactoferrin (agLf) and crossbred goat (cgLf) as estimated by Lowry’s method was found to be 10.94, 12.93 and 11.22mg /L of colostrum respectively. These isolated samples of caprine lactoferrin were found effective to inhibit the growth of both Gram-positive and negative organisms. The assay depicted that the minimum inhibitory concentration (MIC) of mgLf and agLf against E. coli and S. aureus was 275µg/mL and 550µg/mL respectively. The MIC of cgLf against both E. coli and S. aureus was found to be 550µg/mL. The indigenous as well as crossbred goat lactoferrin exhibited the same intensity of antibacterial activity against Gram-positive bacteria. Against Gram-negative organism, lactoferrin of indigenous goats were found to be more potent when compared to the crossbred goat lactoferrin.