Isolation of lactoferrin from milk of different species: Calorimetric and antimicrobial studies

Celia Conesa, Lourdes Sánchez, Carmen Rota, María Dolores Pérez, Miguel Calvo, Sebastien Farnaud, Robert W. Evans

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)131-139
Number of pages9
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume150
Issue number1
DOIs
Publication statusPublished - May 2008
Externally publishedYes

Fingerprint

Lactoferrin
Milk
Earless Seals
New World Camelids
Iron
Camelus
Goats
Escherichia coli
Seals
Hot Temperature
Domestic Sheep
Denaturation
Temperature
Mammals
Protein Stability
Ion Exchange Chromatography
Differential Scanning Calorimetry
Pathogens
Chromatography
Sepharose

Keywords

  • Alpaca LF
  • Antibacterial activity
  • Asian elephant LF
  • Camel LF
  • DSC
  • Goat LF
  • Grey seal milk
  • Human LF
  • Sheep LF

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology

Cite this

Isolation of lactoferrin from milk of different species : Calorimetric and antimicrobial studies. / Conesa, Celia; Sánchez, Lourdes; Rota, Carmen; Pérez, María Dolores; Calvo, Miguel; Farnaud, Sebastien; Evans, Robert W.

In: Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, Vol. 150, No. 1, 05.2008, p. 131-139.

Research output: Contribution to journalArticle

Conesa, Celia ; Sánchez, Lourdes ; Rota, Carmen ; Pérez, María Dolores ; Calvo, Miguel ; Farnaud, Sebastien ; Evans, Robert W. / Isolation of lactoferrin from milk of different species : Calorimetric and antimicrobial studies. In: Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology. 2008 ; Vol. 150, No. 1. pp. 131-139.
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T2 - Calorimetric and antimicrobial studies

AU - Conesa, Celia

AU - Sánchez, Lourdes

AU - Rota, Carmen

AU - Pérez, María Dolores

AU - Calvo, Miguel

AU - Farnaud, Sebastien

AU - Evans, Robert W.

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N2 - 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.

AB - 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.

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KW - Antibacterial activity

KW - Asian elephant LF

KW - Camel LF

KW - DSC

KW - Goat LF

KW - Grey seal milk

KW - Human LF

KW - Sheep LF

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M3 - Article

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EP - 139

JO - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology

JF - Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology

SN - 1096-4959

IS - 1

ER -