ISOLATION AND CHARACTERIZATION OF LACTOFERRIN FROM CARABAO&#039;S (Bubalus bubalis L.) MILK WHEY

Florinia  E. Merca,; N.p.  Roxas,; Amado  A. Angeles,

ISOLATION AND CHARACTERIZATION OF LACTOFERRIN FROM CARABAO'S (Bubalus bubalis L.) MILK WHEY

Amado A. Angeles | Florinia E. Merca | N.p. Roxas

Abstract:

Lactoferrin (cLF) was isolated from carabao (Bubalus bubalis L) milk whey by ammonium sulfate (AS) precipitation and cation exchange chromatography (CEC) using carboxymethyl cellulose (CMC) as cation exchanger. Bovine lactoferrin ELISA kit was used to detect and quantify lactoferrin from the samples. The cLF was assayed for antibacterial activity in vitro. Bovine LF was likewise tested for antibacterial activity for comparison. Carabao LF was eluted at approximately 0.27-0.30 M NaCl with a concentration of 20.05 mg/l and a recovery of 24.97% from the whey sample. SDS-PAGE of cLF showed RF value corresponding to molecular weight of 77,829. Carbohydrate analysis by phenolsulfuric acid test showed carbohydrate content of 0.346% indicating the glycoprotein nature of the cLF. In vitro antibacterial assay showed that cLF can inhibit growth of enteropathogenic E. coli (EPEC) and S. typhimurium with minimum inhibitory concentrations (MIC) of 800 and 1600 μg/ml, respectively. Commercial bovine LF (bLF) on the other hand, had twice the activity of cLF with MICs of 400 and 800 μg/ml for EPEC and S. typhimurium, respectively. Antibacterial activity of cLF and bLF against EPEC and S. typhimurium were stable from pH 3 to 7 and up to 50ºC.