Isolation and identification of 7-oxo-8-aminopelargonic acid, a biotin intermediate

Eisenberg, Max A.; Maseda, Ramon

doi:10.1021/bi00803a014

Cited by 13 publications

(4 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…AOP was determined by using the S. cerevisiae microbiological agar diffusion assay. Different values for the bioactivity of AOP (in the yeast assay) have been reported in the literature [14,15], but we found that racemic AOP was as active as biotin in the concentration range we used (0.1-20 fM), in accordance with Eisenberg & Maseda [14]. This routine discontinuous assay for AOP synthase is quite sensitive, but the manual measurement of growth diameters limits its accuracy (5-10 %).…”

Section: Results and Discussion Syntheses And Assayssupporting

confidence: 81%

The 8-amino-7-oxopelargonate synthase from Bacillus sphaericus. Purification and preliminary characterization of the cloned enzyme overproduced in Escherichia coli

Ploux

Marquet

1992

Biochemical Journal

View full text Add to dashboard Cite

The 8-amino-7-oxopelargonate synthase [6-carboxyhexanoyl-CoA:L-alanine carboxyhexanoyltransferase (decarboxylating); EC 2.3.1.47] from Bacillus sphaericus involved in biotin biosynthesis was purified from an Escherichia coli overproducing strain. The purification afforded an electrophoretically homogeneous enzyme with a specific activity of 0.67 unit/mg. The purified enzyme is a monomer of 41 kDa. N-Terminal sequencing of the first 14 amino acid residues showed complete agreement with the predicted sequence from the bioF gene. The pure enzyme showed the characteristic absorption band (425 nm) of pyridoxal 5'-phosphate-dependent enzymes. Furthermore, the holoenzyme was resolved during an affinity step yielding the inactive apoenzyme, which recovered activity and the 425 nm-absorption band on dialysis against pyridoxal 5'-phosphate. Km values for L-alanine and pimeloyl-CoA were respectively 3 mM and 1 microM.

show abstract

Section: Results and Discussion Syntheses And Assayssupporting

confidence: 81%

The 8-amino-7-oxopelargonate synthase from Bacillus sphaericus. Purification and preliminary characterization of the cloned enzyme overproduced in Escherichia coli

Ploux

Marquet

1992

Biochemical Journal

View full text Add to dashboard Cite

show abstract

“…We are therefore continuing efforts to purify this compound (9) from supernatants of the corresponding bioA auxotroph of E. coli (7). We have also mapped this mutation to a position within 3 cM of the tz locus on chromosome 5 and have begun to test RFLP probes in this region (4) in preparation for gene isolation through chromosome walking.…”

Section: Biotin In Isolated Embryosmentioning

confidence: 99%

Arrested Embryos from the bio1 Auxotroph of Arabidopsis thaliana Contain Reduced Levels of Biotin

Shellhammer¹,

Meinke²

1990

Plant Physiol.

View full text Add to dashboard Cite

The biol auxotroph of Arabidopsis thaliana is a recessive embryonic lethal that forms normal plants in the presence of biotin. The purpose of this study was to determine whether aborted seeds produced by heterozygous plants grown without vitamin supplements contained reduced levels of biotin. Two methods were used to determine the biotin content of mutant and wild-type tissues: streptavidin binding in microtiter plates and growth of the biotin-requiring bacterium Lactobacillus plantarum.Total biotin was measured in extracts prepared from immature seeds prior to desiccation. Aborted seeds produced by heterozygous (biol/BIO1) plants contained some biotin in the maternal seed coat but virtually no detectable biotin in the arrested embryo. This lack of biotin was not observed in arrested embryos from other mutants with similar patterns of abnormal development. These results are consistent with the model that biol tissues are defective in biotin synthesis. The alternative model of increased degradation is inconsistent with the recessive nature of the mutation and the ability of rescued plants to continue growing for several weeks following removal of supplemental biotin.

show abstract

“…Although biotin contains a nitrogen atom in the Yas well as the 3'-position, the need to distinguish between these two atoms in the mass spectrum of biotin has been eliminated by the findings of previous investigations. Iwahara et al (1966), Eisenberg &Star (1968), andEisenberg &Maseda (1969) have shown that L-alanine condenses with pimeloyl coenzyme A to form 7-keto-8-aminopelargonic acid. The l'-nitrogen atom of biotin is therefore contributed by L-alanine, and consequently, any significant incorporation of 15N seen in the experiments presented herein must be in the 3'-nitrogen.…”

Section: Resultsmentioning

confidence: 99%

Determination of the metabolic origins of the sulfur and 3'-nitrogen atoms in biotin of Escherichia coli by mass spectrometry

DeMoll¹,

White²,

Shive³

1984

Biochemistry

View full text Add to dashboard Cite

Two steps in the biosynthesis of biotin in Escherichia coli, incorporation of the nitrogen atom of methionine into 7-keto-8-aminopelargonic acid and of the sulfur atom into dethiobiotin, were examined. Sulfur and nitrogen metabolism were monitored by gas chromatography-mass spectrometry of volatile derivatives of internal (protein-bound) amino acids and excreted biotin. We were able to show that internal cysteine and excreted biotin were labeled to the same extent with 34S from either of two exogenous sulfur sources, 34SO4(2)-or L-[sulfane-34S]thiocystine. Internal methionine was eliminated from consideration, while cysteine, or possibly a closely related intermediate, was implicated as providing the sulfur atom for biotin biosynthesis. Also, in experiments designed to follow the metabolism of the nitrogen atom of methionine, it was found that biotin excreted into the culture medium by this organism grown with 95 atom % [15N]methionine contained greater than 70 atom % excess 15N in one of the nitrogens over that obtained from cultures grown with methionine of natural abundance 15N. These results provide evidence for the direct transfer of the methionine nitrogen as the role of S-adenosylmethionine in the conversion of 7-keto-8-aminopelargonic acid to 7,8-diaminopelargonic acid.

show abstract

Isolation and identification of 7-oxo-8-aminopelargonic acid, a biotin intermediate

Cited by 13 publications

References 8 publications

The 8-amino-7-oxopelargonate synthase from Bacillus sphaericus. Purification and preliminary characterization of the cloned enzyme overproduced in Escherichia coli

The 8-amino-7-oxopelargonate synthase from Bacillus sphaericus. Purification and preliminary characterization of the cloned enzyme overproduced in Escherichia coli

Arrested Embryos from the bio1 Auxotroph of Arabidopsis thaliana Contain Reduced Levels of Biotin

Determination of the metabolic origins of the sulfur and 3'-nitrogen atoms in biotin of Escherichia coli by mass spectrometry

Contact Info

Product

Resources

About