A common precursor for the three subunits ofL-glutamate oxidase encoded bygoxgene fromStreptomyces platensisNTU3304

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 alpha, beta, and gamma, 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 alpha-gamma-beta. 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.

Interaction of l-glutamate oxidase with triazine dyes: selection of ligands for affinity chromatography

“…This K m value is in agreement with that reported for Streptomyces endus enzyme [1] but differs significantly to that reported for the Streptomyces sp. 80-5 enzyme (0.21 mM) [2].…”

“…The biological significance of the majority of l-amino specific oxidases remains uncertain due to the lack of sufficient data [1]. Streptomyces l-glutamate oxidase (GOX) is an extracellular heterotrimeric enzyme consisting of ␣, ␤ and ␥ subunits of molecular masses equal to 39, 19 and 16 kDa, respectively [1][2][3].…”

“…Streptomyces l-glutamate oxidase (GOX) is an extracellular heterotrimeric enzyme consisting of ␣, ␤ and ␥ subunits of molecular masses equal to 39, 19 and 16 kDa, respectively [1][2][3]. The enzyme attracts attention because of its biotechnological applications.…”

“…This is mainly due to its limited supply in a native form and unavailability in a recombinant form. To these problems, one must add the laborious and inefficient purification procedures available so far, involving four steps of which three being chromatographic [1,2,7], leading to modest enzyme recovery.…”

Isolation, purification, and characterization of L-glutamate oxidase from Streptomyces sp. 18G

“…X-119-6 had a molecular weight of approximately 140 kDa and consisted of 3 types of subunits, a, b and g with molecular weights of approximately 44, 19 and 9 kDa, respectively. Recently, another GLOD with three subunits was obtained from S. platensis NTU3304 (Chen et al 2001). The pI of GLOD from Streptomyces sp.…”