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Structural characteristic, pH and thermal stabilities of apo and holo forms of caprine and bovine lactoferrins

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Abstract

Apo and holo forms of lactoferrin (LF) from caprine and bovine species have been characterized and compared with regard to the structural stability determined by thermal denaturation temperature values (T m), at pH 2.0–8.0. The bovine lactoferrin (bLF) showed highest thermal stability with a T m of 90 ± 1°C at pH 7.0 whereas caprine lactoferrin (cLF) showed a lower T m value 68 ± 1°C. The holo form was much more stable than the apo form for the bLF as compared to cLF. When pH was gradually reduced to 3.0, the T m values of both holo bLF and holo cLF were reduced showing T m values of 49 ± 1 and 40 ± 1°C, respectively. Both apo and holo forms of cLF and bLF were found to be most stable at pH 7.0. A significant loss in the iron content of both holo and apo forms of the cLF and bLF was observed when pH was decreased from 7.0 to 2.0. At the same time a gradual unfolding of the apo and holo forms of both cLF and bLF was shown by maximum exposure of hydrophobic regions at pH 3.0. This was supported with a loss in α-helix structure together with an increase in the content of unordered (aperiodic) structure, while β structure seemed unchanged at all pH values. Since LF is used today as fortifier in many products, like infant formulas and exerts many biological functions in human, the structural changes, iron binding and release affected by pH and thermal denaturation temperature are important factors to be clarified for more than the bovine species.

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Abbreviations

LF:

Lactoferrin

apo LF:

Apo lactoferrin

holo LF:

Holo lactoferrin

cLF:

Caprine lactoferrin

bLF:

Bovine lactoferrin

Trp:

Tryptophan

CD:

Circular dichroism

far-UV CD:

Far-ultraviolet circular dichroism

T m :

Thermal denaturation temperature

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Acknowledgments

The author gratefully acknowledges Daniel Krowarsch, Faculty of Biotechnology, University of Wroclaw, Poland for providing circular dichroism facility and technical assistance. Further, the author is thankful to Tor Bruun, Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences for assistance during protein purification.

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Authors and Affiliations

  1. Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Aas, 1432, Norway

    Ashoka Sreedhara, Ragnar Flengsrud, Thor Langsrud & Gerd Elisabeth Vegarud

  2. Department of Protein Chemistry and Technology, Central Food Technological Research Institute (A Constituent Laboratory of CSIR, Delhi), Mysore, 570 020, India

    Ashoka Sreedhara, Purnima Kaul & Vishweshwaraiah Prakash

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  1. Ashoka Sreedhara
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  2. Ragnar Flengsrud
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  3. Thor Langsrud
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  4. Purnima Kaul
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  5. Vishweshwaraiah Prakash
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  6. Gerd Elisabeth Vegarud
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Correspondence to Ashoka Sreedhara.

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Sreedhara, A., Flengsrud, R., Langsrud, T. et al. Structural characteristic, pH and thermal stabilities of apo and holo forms of caprine and bovine lactoferrins. Biometals 23, 1159–1170 (2010). https://doi.org/10.1007/s10534-010-9366-5

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  • DOI: https://doi.org/10.1007/s10534-010-9366-5

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