A common precursor for the three subunits of L-glutamate oxidase encoded by gox gene from Streptomyces platensis NTU3304

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Abstract:

A segment of DNA containing the L-glutamate oxidase (gox) gene from Streptomyces platensis NTU3304 was cloned. The entire nucleotide sequence of the protein-coding portion consisting of 2130 bp (710 codons, including AUG and UGA) of the cloned DNA fragment was determined. The gox gene contained only one open reading frame (ORF) which coded for a 78-kDa polypeptide, the precursor of active extracellular Gox. Mature Gox is composed of three subunits, designated as α, , and , with molecular masses of 39, 19, and 16 kDa, respectively. Analyses of the N-terminal amino acid sequences of the subunits revealed that the order of subunits in the precursor polypeptide encoded by the ORF, from N-terminus to C-terminus, is α––. The presence of the flavin adenine dinucleotide (FAD)-binding motif place Gox as a member of the flavoenzyme family. Furthermore, a negative effect of glucose on the biosynthesis of Gox was observed when it was used as carbon source.Key words: L-glutamate oxidase, gox gene, signal peptide, DNA sequence, flavoenzyme, pIJ702 vector.

Nous avons cloné un segment d'ADN de Streptomyces platensis NTU3304 contenant le gène de la L-glutamate oxidase (gox). La séquence complète de nucléotides de la portion codante de l'ADN cloné, représentant 2130 pb (710 codons, incluant les AUG et UGA), fut déterminée. Le gène gox contenait seulement un cadre de lecture ouvert (CLO) qui codait un polypeptide de 78-kDa, le précurseur de la protéine Gox extracellulaire. La protéine Gox mature est formée de trois sous-unités, nommées α, , et , ayant des masses moléculaires respectives de 39, 19, et 16 kDa. Les analyses des séquence amino-terminales des acides aminés des sous-unités ont révélé que l'ordre des sous-unités, telle que retrouvée dans le polypeptide précurseur codé par le CLO, est α––, de la terminaison aminée à la terminaison carboxy. Le présence du motif de liaison à la flavine adénine dinucléotide (FAD) place Gox dans la famille des flavoenzymes. Dans une autre expérience, le glucose a eu un effet négatif sur la biosynthèse de Gox lorsqu'il était utilisé en tant que source de carbone.Mots clés : L-glutamate oxidase, gène gox, peptide de signal, séquence d'ADN, flavoenzyme, vecteur pIJ702.[Traduit par la Rédaction]

Keywords: DNA sequence; L-glutamate oxidase; flavoenzyme; gox gene; gène gox; pIJ702 vector; peptide de signal; signal peptide; séquence d'ADN; vecteur pIJ702

Document Type: Research Article

Publication date: March 1, 2001

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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