Analysis of o-phthalaldehyde derivatives of acidic and polar amino acids in fermentation broths by high-performance liquid chromatography

White, Robert L.; DeMarco, Alphonse C.; Smith, Kevin C.

doi:10.1016/s0021-9673(01)93147-4

Cited by 11 publications

(8 citation statements)

References 16 publications

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“…Amino acid concentrations in Fusobacterium cultures were monitored by HPLC analysis of fluorescent isoindole derivatives (White et al, 1989). After incubation of cultures supplied with racemic amino acids the residual amino acid concentration was close to either 0, 50 or 100% of the initial concentration, indicating complete, partial or insignificant utilization of each enantiomer.…”

Section: Resultsmentioning

confidence: 99%

“…After 1rain at ambient temperature, sodium acetate solution (120/d, 0.1M, pH 6.5) was added, and a 20-~tl portion was injected onto a Beckman Ultrasphere ODS column (5/~m, 45 × 4.6ram). Separations were achieved at a total flow rate of 2.5 ml/min using gradients formed between sodium acetate (0.1M, pH 6.5 with 3M HC1)-methanol-THF (900:95:5) and methanol (White et al, 1989). Alanine, a-aminoadipic acid, arginine, aspartic acid, glutamic acid, glutamine, isoleucine, leucine, methionine, serine, tyrosine, and valine were analyzed using a gradient of composition (rain, % methanol): 0.0, 0; 0.…”

Section: Amino Acid Analysis By Hplcmentioning

confidence: 99%

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Utilization ofd-amino acids byFusobacterium nucleatum andFusobacterium varium

1999

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The utilization of D- and L-amino acids with acidic, basic or polar side chains was demonstrated by HPLC. Two species of the anaerobe Fusobacterium utilized D-lysine and the L isomers of glutamate, glutamine, histidine, lysine and serine. Only F. varium used L-arginine, D-glutamate and D-serine as substrates, whereas F. nucleatum specifically utilized D-histidine and D-glutamine. D-Glutamate accumulated in F. nucleatum cultures supplemented with D-glutamine, and ornithine was detected when either DL- or L-arginine was included in F. varium cultures. Based on literature precedents, D-glutamate and D-histidine are isomerized to their L isomers prior to degradation, but separate catabolic pathways are possible for each enantiomer of lysine and serine.

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Section: Resultsmentioning

confidence: 99%

Section: Amino Acid Analysis By Hplcmentioning

confidence: 99%

Utilization ofd-amino acids byFusobacterium nucleatum andFusobacterium varium

1999

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“…Although HON is readily detected in culture supernatants by hplc using precolumn derivatization with ophthalaldehyde (7), neither HON nor any other metabolite was detected by this method in culture supernatants of mutants L127, LI38, and L167 over incubation for four days on casein-starch medium. Because this derivatization method is selective for primary amines, and intermediates of HON biosynthesis need not be free amino acids (5), a bioassay-directed purification of the metabolite(s) accumulated by mutant LI38 was undertaken.…”

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confidence: 79%

Isolation of N-Acetyl-3,4-dihydroxy-L-phenylalanine from Streptomyces akiyoshiensis

Smith¹,

White²,

Le³

et al. 1995

J. Nat. Prod.

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Acetyl-3,4-dihydroxy-L-phenylalanine (N-acetyl-L-DOPA) was isolated from cultures of Streptomyces akiyoshiensis. The structure was deduced from spectroscopic data and confirmed by comparison to a standard sample synthesized from acetic anhydride and L-DOPA.N-Acetyl-L-DOPA does not appear to be involved in the biosynthesis of 5-hydroxy-4-oxo-Lnorvaline (HON), the major metabolite of S. akiyoshiensis.A non-protein amino acid, 5-hydroxy-4-oxo-L-norvaline (HON), which possesses antitubercular (1) and antifungal (2) properties is produced by the soil bacterium Streptomyces akiyoshiensis (Streptomycetaceae) (3,4). Aspartate and acetate have been identified as the biosynthetic precursors of HON by isotopic feeding experiments ( 5), and ongoing investigations to obtain additional biosynthetic information are focused on mutants of S. akiyoshiensis unable to synthesize HON. Three groups of nonauxotrophic mutants have been obtained, and the relative order of the blocked steps has been determined by cross-feeding experiments; for example, culture supernatant obtained from mutant LI38 stimulates HON synthesis in mutants LI27 and LI 67 (6). Inasmuch as this effect could be explained by the accumulation in mutant LI 38 cultures of a biosynthetic intermediate (or a related compound) that is con-

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“…Diluted culture broth or cell extract (20 kL) was mixed with an internal standard solution (20 kL, cysteic acid, 50 nmoVmL) and derivatized using a commercial o-phthalaldehyde reagent (Beckman Fluor-R, Beckman Instruments. Mississauga, Ont., or Sigma o-phthalaldehyde reagent, Sigma Chemical Company, St. Louis, Mo., U.S.A.) according to published conditions (13). The fluorescent OPA-derivatives were separated by gradient elution on a reversed-phase column as described previously (Figure l b in ref.…”

Section: Hplc Analysis Of Amino Acidsmentioning

confidence: 99%

“…by the observation that efficient aeration of S. akiyoshiensis culsince the claisen condensation in pepstatin biosynthesis tures is required for HON production (3 I), but the oxidation and occurs at the c-terminal leucine residue of a peptide (40) and Nde~arbox~lation steps needed to convert the product of the conacetylglutamate is an intermediate in the b i o s~~t h e s i s of ornidensation step (13) to HON (1) could occur simultaneously or in thine from glutamate (5 I), it is conceivable that a derivative of two alternative sequences (Scheme 3). There is some precedent aspartic acid or an aspartic acid residue in a peptide is involved for each of the latter.…”

mentioning

confidence: 99%

Biosynthesis of 5-hydroxy-4-oxo-L-norvaline in Streptomyces akiyoshiensis

White¹,

Smith²,

DeMarco³

1994

Can. J. Chem.

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The biosynthesis of 5-hydroxy-4-0x0-L-norvaline (HON) in Streptomyces akiyoshiensis has been investigated using I3c-labelled substrates. Incorporations of I3c label from sodium [I-"c]-, [2-I3c]-, and [1,2-13C2]acetate indicated that HON was formed from a four-carbon compound derived from the citric acid cycle and the methyl carbon of acetate. Feeding experiments using D L -[~-'~~I -and ~~-[ 2 -'~~, '~~] a s~a r t a t e demonstrated that aspartate served as the four-carbon precursor to HON. Both enantiomers of aspartate were metabolized by S. akiyoshiensis, but the D isomer was consumed at a slower rate. The distribution of I3C label in the intracellular L-glutamic acid isolated in these feeding experiments is consistent with the operation of the citric acid cycle in S. akiyoshiensis. A biosynthetic hypothesis that involves a condensation reaction between acetyl or malonyl CoA and the P-carboxyl group of aspartate, and subsequent oxidative decarboxylation, is proposed to account for the incorporation results. An analogous condensation step has been proposed for the biosynthesis of other natural products, including the carbapenem antibiotics. ~~-[ 2 -~~C ,~~~]~s~a r t a t e was synthesized from [Traduit par la rCdaction]

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Analysis of o-phthalaldehyde derivatives of acidic and polar amino acids in fermentation broths by high-performance liquid chromatography

Cited by 11 publications

References 16 publications

Utilization ofd-amino acids byFusobacterium nucleatum andFusobacterium varium

Utilization ofd-amino acids byFusobacterium nucleatum andFusobacterium varium

Isolation of N-Acetyl-3,4-dihydroxy-L-phenylalanine from Streptomyces akiyoshiensis

Biosynthesis of 5-hydroxy-4-oxo-L-norvaline in Streptomyces akiyoshiensis

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