Use of stable isotopes to measure de novo synthesis and turnover of amino acid-C and -N in mixed micro-organisms from the sheep rumen in vitro

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dc.contributor.authorAtasoglu, C.
dc.contributor.authorGuliye, A.Y.
dc.contributor.authorWallace, R.J.
dc.date.accessioned2025-01-27T18:55:57Z
dc.date.available2025-01-27T18:55:57Z
dc.date.issued2004
dc.departmentÇanakkale Onsekiz Mart Üniversitesi
dc.description.abstractProtein synthesis and turnover in ruminal micro-organisms were assessed by stable-isotope methods in order to follow independently the fate of amino acid (AA)-C and -N in different AA. Rumen fluid taken from sheep receiving a grass hay-concentrate diet were strained and incubated in vitro with starch-cellobiose-xylose in the presence of NH3 and 5 g algal protein hydrolysate (APH)/l, in incubations where the labels were 15NH3, [15N]APH or [13C]APH. Total 15N incorporation was calculated from separate incubations with 15NH3 and [15N]APH, and net N synthesis from the increase in AA in protein-bound material. The large difference between total and net AA synthesis indicated that substantial turnover of microbial protein occurred, averaging 3.5 %/h. Soluble AA-N was incorporated on average more extensively than soluble AA-C (70 v. 50 % respectively, P=0.001); however, incorporation of individual AA varied. Ninety percent of phenylalanine-C was derived from the C-skeleton of soluble AA, whereas the incorporation of phenylalanine-N was 72 %. In contrast, only 15 % aspartate-C + asparagine-C was incorporated, while 45 % aspartate-N + asparagine-N was incorporated. Deconvolution analysis of mass spectra indicated substantial exchange of carboxyl groups in several AA before incorporation and a condensation of unidentified C2 and C4 intermediates during isoleucine metabolism. The present results demonstrate that differential labelling with stable isotopes is a way in which fluxes of AA synthesis and degradation, their biosynthetic routes, and separate fates of AA-C and -N can be determined in a mixed microbial population. © The Authors 2004.
dc.description.sponsorshipScottish Executive Environment and Rural Affairs Department
dc.identifier.doi10.1079/BJN20031040
dc.identifier.endpage261
dc.identifier.issn0007-1145
dc.identifier.issue2
dc.identifier.pmid14756911
dc.identifier.scopus2-s2.0-1342346115
dc.identifier.scopusqualityQ1
dc.identifier.startpage253
dc.identifier.urihttps://doi.org/10.1079/BJN20031040
dc.identifier.urihttps://hdl.handle.net/20.500.12428/12780
dc.identifier.volume91
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.relation.ispartofBritish Journal of Nutrition
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/openAccess
dc.snmzKA_Scopus_20250125
dc.subjectAmino acids; Carbon skeletons; Rumen; Sheep
dc.titleUse of stable isotopes to measure de novo synthesis and turnover of amino acid-C and -N in mixed micro-organisms from the sheep rumen in vitro
dc.typeArticle

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