Development of a method to assess binding of astaxanthin to Atlantic salmon Salmo salar L. muscle proteins

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DOIResolve DOI: http://doi.org/10.1111/j.1365-2109.2004.01205.x
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TypeArticle
Journal titleAquaculture Research
Volume36
Issue4
Pages336343; # of pages: 8
Subjectanalytical techniques; methodology Muscles; chemical reactions; colour; chromatographic techniques; filtration Fish culture; actin; biochemical analysis; proteins; aquaculture techniques
AbstractSeveral methods were examined to characterize the binding between astaxanthin and salmon muscle protein(s) in order to provide tools for evaluation of the role of muscle proteins on astaxanthin retention in Atlantic salmon Salmo salar L. flesh. The methods included gel filtration chromatography, displacement of a hydrophobic probe and ultrafiltration. With gel filtration chromatography, aggregation of astaxanthin under the experimental conditions was a major problem for the separation of bound astaxanthin from free astaxanthin because the apparent molecular weight of aggregated astaxanthin or astaxanthin micelles was in the range of protein–astaxanthin complexes. Displacement of the fluorescent probe 8-anilino-1-naphthalenesulphonate (ANS) was not effective as astaxanthin quenched the fluorophore so that displacement could not be observed. An ultrafiltration method was developed using 200-mM sodium cholate for dispersion of astaxanthin aggregates. This allowed unbound astaxanthin to be separated from bound astaxanthin using a 30-kDa filter. After salmon muscle proteins were solubilized in different fractions by sequential extraction using low ionic strength solutions, the astaxanthin binding of different fractions was assessed using the ultrafiltration method. The significant difference (P<0.05) observed in the astaxanthin binding of the various fractions suggests an application of this assay to detect differences in affinity of proteins for astaxanthin. The results also suggest that proteins other than actomyosin or actin can bind astaxanthin in Atlantic salmon flesh. This method can be used for the identification of astaxanthin-binding proteins in salmon flesh and other tissues.
Publication date
Copyright noticeCopyright © 2005, Blackwell Publishing Ltd.
LanguageEnglish
AffiliationNRC Institute for Marine Biosciences; National Research Council Canada; Industrial Research Assistance Program; Aquatic and Crop Resource Development
Peer reviewedYes
NRC number1670
NPARC number3538359
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Record identifier09c2b741-0ea1-4c19-a561-571ffc6e2602
Record created2009-03-01
Record modified2016-05-09
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