The stable isotope composition of amino acids refers to the abundance of heavy and light non-radioactive isotopes of carbon (13C and 12C), nitrogen (15N and 14N), and other elements within these molecules. Amino acids are the building blocks of proteins. They are synthesized from alpha-keto acid precursors that are in turn intermediates of several different pathways in central metabolism. Carbon skeletons from these diverse sources are further modified before transamination, the addition of an amino group that completes amino acid biosynthesis. Bonds to heavy isotopes are stronger than bonds to light isotopes, making reactions involving heavier isotopes proceed slightly slower in most cases. This phenomenon, known as a kinetic isotope effect, gives rise to isotopic differences between reactants and products that can be detected using isotope ratio mass spectrometry. Amino acids are synthesized via a variety of pathways with reactions containing different, unknown isotope effects. Because of this, the 13C content of amino acid carbon skeletons varies considerably between the amino acids. There is also an isotope effect associated with transamination, which is apparent from the abundance of 15N in some amino acids. Because of these properties, amino acid isotopes record useful information about the organisms that produce them. Variations in metabolism between different taxonomical groups give rise to characteristic patterns of 13C enrichment in their amino acids. This allows the sources of carbon in food webs to be identified. The isotope effect associated with transamination also makes amino acid nitrogen isotopes a useful tool to study the structure of food webs. Repeated transamination by consumers results in a predictable increase in the abundance of 15N as amino acids are transferred up food chains. Together, these application, among others in ecology, demonstrate the utility of stable isotopes as tracers of environmental processes that are difficult to measure directly.
Isotopic fractionation in reaction networks To explain the wide range of isotopic compositions observed among the amino acids, it is necessary to consider how isotopes are sorted between starting materials, intermediates, and products in reaction networks. Amino acid biosynthesis pathways contain both reversible and irreversible reactions, as well as branch points where one intermediate can react to form two different products. The following examples adapted from Hayes (2001) illustrate the isotopic consequences of these network structures.
Linear irreversible network In the following reaction network, A is irreversibly converted to an intermediate B, which irreversibly reacts to form C.
A → δ b , α b / A ϕ a b B → δ c , α c / B ϕ b c C {\displaystyle {\ce {A->[{\phi _{ab}}][{\delta _{b},\alpha _{b/A}}]B->[{\phi _{bc}}][{\delta _{c},\alpha _{c/B}}]C}}}
The pools of A, B, and C have delta values defined as δA, δB, and δC respectively. These values are related to the ratio of heavy to light isotopes in each pool, and are the conventional means by which scientists express the isotopic composition of materials. Importantly, δB is distinct from δb listed on the diagram, as δb is the isotopic composition of B produced from A before it mixes with the pool of B. The isotopic compositions of the pools and products are related through fractionation factors that reflect the kinetic isotope effects (KIEs) associated with each reaction. For A → B,
α b / A ≡ δ b + 1 δ A + 1 {\displaystyle \alpha _{b/A}\equiv {\frac {\delta _{b}+1}{\delta _{A}+1}}}
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![Stable isotope composition of amino acids: Some amino acids have carbon isotope compositions that reflect the organism that produced them. The x axis is the difference in δ13C between isoleucine and leucine, while the y axis shows this difference for isolecuine and lysine. There are clear clusters of points corresponding to bacteria, fungi, and plants. Figure adapted from.[3]](https://upload.wikimedia.org/wikipedia/commons/thumb/1/10/Isoleucine_vs_leucine_and_lysine_d13C_for_amino_acids_from_different_organisms.svg/500px-Isoleucine_vs_leucine_and_lysine_d13C_for_amino_acids_from_different_organisms.svg.png?utm_source=en.wikipedia.org&utm_campaign=parser&utm_content=thumbnail)
