Phanerochaete chrysosporium glyoxal oxidase is encoded by two allelic variants: structure, genomic organization, and heterologous expression of glx1 and glx2.
A cDNA clone (glx-2c) encoding glyoxal oxidase (GLOX) was isolated from a Phanerochaete chrysosporium lambda gt11 library, and its nucleotide sequence was shown to be distinct from that of the previously described clone glx-1c (P. J. Kersten and D. Cullen, Proc. Natl. Acad. Sci. USA 90:7411-7413, 1993). Genomic clones corresponding to both cDNAs were also isolated and sequenced. overall nucleotide sequence identity was 98%, and the predicted proteins differed by a single residue: Lys-308<==>Thr-308. Analyses of parental dikaryotic strain BKM-F-1767 and homokaryotic progeny firmly established allelism for these structural variants. Southern blots of pulsed-field gels localized the GLOX gene (glx) to a dimorphic chromosome separate from the peroxidase and cellobiohydrolase genes of P. chrysosporium. Controlled expression of active GLOX was obtained from Aspergillus nidulans transformants when glx-1c was fused to the promoter and secretion signal of the A. niger glucoamylase gene. The GLOX isozyme corresponding to glx-2c was also efficiently secreted by A. nidulans following site-specific mutagenesis of the expression vector at codon 308 of glx-1c.
Kersten
P J
PJ
Forest Products Laboratory, U.S. Department of Agriculture Forest Service, Madison, Wisconsin, USA.
Witek
C
C
vanden Wymelenberg
A
A
Cullen
D
D
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D000483:Alleles; D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics; D001483:Base Sequence; D015139:Blotting, Southern; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D005658:Fungi / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D005800:Genes, Fungal / Q000235:genetics*; D008040:Genetic Linkage; D014644:Genetic Variation; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis
D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; C052371:glyoxal oxidase