+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Compound-specific nitrogen isotope analysis of D-alanine, L-alanine, and valine: application of diastereomer separation to delta15N and microbial peptidoglycan studies

Compound-specific nitrogen isotope analysis of D-alanine, L-alanine, and valine: application of diastereomer separation to delta15N and microbial peptidoglycan studies

Analytical Chemistry 81(1): 394-399

We have developed an analytical method to determine the compound-specific nitrogen isotope compositions of individual amino acid enantiomers using gas chromatography/combustion/isotope ratio mass spectrometry. A novel derivatization of amino acid diastereomers by optically active (R)-(-)-2-butanol or (S)-(+)-2-butanol offers two advantages for nitrogen isotope analysis. First, chromatographic chiral separation can be achieved without the use of chiral stationary-phase columns. Second, the elution order of these compounds on the chromatogram can be switched by a designated esterification reaction. We applied the method to the compound-specific nitrogen isotope analysis of D- and L-alanine in a peptidoglycan derived from the cell walls of cultured bacteria (Firmicutes and Actinobacteria; Enterococcus faecalis, Staphylococcus aureus, Staphylococcus staphylolyticus, Lactobacillus acidophilus, Bacillus subtilis, Micrococcus luteus, and Streptomyces sp.), natural whole bacterial cells (Bacillus subtilis var. natto), (pseudo)-peptidoglycan from archaea (Methanobacterium sp.), and cell wall from eukaryota (Saccharomyces cerevisiae). We observed statistically significant differences in nitrogen isotopic compositions; e.g., delta15N ( per thousand vs air) in Staphylococcus staphylolyticus for d-alanine (19.2 +/- 0.5 per thousand, n = 4) and L-alanine (21.3 +/- 0.8 per thousand, n = 4) and in Bacillus subtilis for D-alanine (6.2 +/- 0.2 per thousand, n = 3) and L-alanine (8.2 +/- 0.4 per thousand, n = 3). These results suggest that enzymatic reaction pathways, including the alanine racemase reaction, produce a nitrogen isotopic difference in amino acid enantiomers, resulting in 15N-depleted D-alanine. This method is expected to facilitate compound-specific nitrogen isotope studies of amino acid stereoisomers.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 052269271

Download citation: RISBibTeXText

PMID: 19061396

DOI: 10.1021/ac802077v

Related references

Roles of Mycobacterium smegmatis D-alanine:D-alanine ligase and D-alanine racemase in the mechanisms of action of and resistance to the peptidoglycan inhibitor D-cycloserine. Antimicrobial Agents and ChemoTherapy 47(1): 283-291, 2002

Stereospecificity of phenyl alanine plasma kinetics and hydroxylation in man following oral application of a stable isotope labeled pseudo racemic mixture of l phenyl alanine and d phenyl alanine. Clinica Chimica Acta 128(2-3): 181-198, 1983

Solid phase synthesis of 1 asparagine 5 valine 8 alanine angiotensin ii and 1 hydantoic acid 5 valine 8 alanine angiotensin ii. Bioorganicheskaya Khimiya 1(9): 1257-1262, 1975

Thermal variations, between 1 and 3000 degrees K, of the specific heat of L-alanine, tri(L-alanine) and poly(L-alanine) for the alpha and beta forms. Biopolymers 14(4): 801-823, 1975

Isotope labeling studies on the formation of multiple addition products of alanine in the pyrolysis residue of glucose/alanine mixtures by high-resolution ESI-TOF-MS. Journal of Agricultural and Food Chemistry 59(21): 11629-11634, 2012

Tyrosine 265 of alanine racemase serves as a base abstracting alpha-hydrogen from L-alanine: the counterpart residue to lysine 39 specific to D-alanine. Journal of Biochemistry 126(4): 781-786, 1999

A study of conformational stability of poly(L-alanine), poly(L-valine), and poly(L-alanine)/poly(L-valine) blends in the solid state by (13)C cross-polarization/magic angle spinning NMR. Biopolymers 64(1): 26-33, 2002

Peptidoglycan biosynthesis in Escherichia coli: variations in the metabolism of alanine and D-alanyl-D-alanine. Journal of General Microbiology 138 Pt 8: 1751-1757, 1992

beta-Chloro-L-alanine inhibition of the Escherichia coli alanine-valine transaminase. Journal of Bacteriology 164(3): 1350-1352, 1985

B-Chloro- L-alanine inhibition of the Escherichia coli alanine-valine transaminase. Journal of Bacteriology 164: 50-52, 1985

Growth inhibition of Escherichia coli W by D-norvalyl-D-alanine: an analogue of D-alanine in position 4 of the peptide subunit of peptidoglycan. Antimicrobial Agents and ChemoTherapy 11(4): 638-644, 1977

Inert free radicals as spin labels part 2 reactions of 4 hydroxytetradecachlorotriphenylmethyl radical with alanine phenyl alanine valine and proline. Tetrahedron Letters (5): 479-480, 1978

Germination initiation and inhibitory activities of l alanine and d alanine analogs for bacillus subtilis spores modification of methyl group of l alanine and d alanine. Microbiology & Immunology 29(3): 229-242, 1985

Angiotensin-II Analogues. I: Synthesis and incorporation of the halogenated amino acids 3-(4-iodophenyl)alanine, 3-(3,5-dibromo-4-chlorophenyl)alanine, 3-(3,4,5-tribromophenyl)alanine, and 3-(2,3,4,5,6-pentabromophenyl)alanine. Helvetica Chimica Acta 66(3): 960-970, 1983

Stereoselective synthesis of stable isotope-labeled L-alpha-amino acids Electrophilic amination of Oppolzers acyl sultams in the synthesis of L- alanine, L- valine, L- leucine, L- phenylalanine and L- valine. Journal of Labelled Compounds & Radiopharmaceuticals 38(3): 239-248, 1996