Isolation of lactoferrin from milk of different species: calorimetric and antimicrobial studies.
Lactoferrin (LF) is an iron-binding glycoprotein found in different biological fluids of mammals and in neutrophils. It has been proposed to be involved in many functions, including protection from pathogens. In this work, purification of lactoferrin using an ion-exchange chromatography (SP-Sepharose) was attempted for the milk of the following animals: sheep (Ovis aries), goat (Capra hircus), camel (Camelus bactrianus), alpaca (Lama pacos), elephant (Elephas maximus) and grey seal (Halichoerus grypus), as well as human (Homo sapiens). Lactoferrin was identified in all the milks apart from that from grey seal. The thermal stability of the purified lactoferrins, in their native and iron-saturated forms, was studied by differential scanning calorimetry (DSC). Maximum temperature, onset temperature and enthalpy change of denaturation were higher when lactoferrins were saturated with iron than in their native form, indicating an increase in the stability of the protein structure upon iron-binding. Human lactoferrin was found to be the most heat-resistant and the other lactoferrins presented different degrees of thermoresistance, that of elephant being the least resistant. The antimicrobial activity of the different isolated lactoferrins was investigated against Escherichia coli 0157:H7. The minimal inhibitory concentrations (MICs) were determined by measuring the absorbance at 620 nm. The minimum bactericidal concentrations (MBCs) were also measured and it was found that camel lactoferrin was the most active lactoferrin against E. coli 0157:H7, whereas alpaca and human lactoferrins were the least active.
Conesa
Celia
C
Tecnología de los Alimentos, Facultad de Veterinaria, Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain.
Sánchez
Lourdes
L
Rota
Carmen
C
Pérez
María-Dolores
MD
Calvo
Miguel
M
Farnaud
Sebastien
S
Evans
Robert W
RW
D000818:Animals; D000890:Anti-Infective Agents / Q000302:isolation & purification* / Q000494:pharmacology*; D002152:Calorimetry, Differential Scanning; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000187:drug effects; D006801:Humans; D005779:Immunodiffusion; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000302:isolation & purification* / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D008892:Milk / Q000737:chemistry*; D013045:Species Specificity; D013696:Temperature
D000890:Anti-Infective Agents; D007501:Iron; D007781:Lactoferrin