Cellobiose quinone oxidoreductase from the white rot fungus Phanerochaete chrysosporium is produced by intracellular proteolysis of cellobiose dehydrogenase

The fungus Phanerochaete chrysosporium was grown in a 10-l automatic fermenter using cellobiose as carbon source to monitor the induction of cellobiose dehydrogenase (CDH) and cellobiose quinone oxidoreductase (CBQ) enzymes, and to search for tentative cbq and cdh genes and their transcriptional pro...

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Bibliographic Details
Published in:Biochimica et biophysica acta 2002-06, Vol.1576 (1), p.15-22
Main Authors: Raı́ces, Manuel, Montesino, Raquel, Cremata, José, Garcı́a, Bianca, Perdomo, Walmer, Szabó, István, Henriksson, Gunnar, Hallberg, B.Martin, Pettersson, Göran, Johansson, Gunnar
Format: Article
Language:English
Subjects:
Alleles
Blotting, Southern
Blotting, Western
Carbohydrate Dehydrogenases - biosynthesis
Carbohydrate Dehydrogenases - chemistry
Cellobiose dehydrogenase
Cellobiose quinone oxidoreductase
cellulose
Chromatography, High Pressure Liquid
domains
Expression
Fungal Proteins - biosynthesis
Fungal Proteins - chemistry
genes
lignin
oxidase
Phanerochaete - genetics
Phanerochaete - metabolism
Phanerochaete chrysosporium
proteins
Proteolysis
purification
Serine Endopeptidases
sporotrichum-pulverulentum
Transcription, Genetic
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Description
Summary:The fungus Phanerochaete chrysosporium was grown in a 10-l automatic fermenter using cellobiose as carbon source to monitor the induction of cellobiose dehydrogenase (CDH) and cellobiose quinone oxidoreductase (CBQ) enzymes, and to search for tentative cbq and cdh genes and their transcriptional products. After 24 h of induction, CDH was detected in the culture supernatant and a protein was recognized by a specific anti-CDH polyclonal antibody in the sonicated biomass. Northern blot experiments performed with several fungal RNA samples showed, after 24 h of induction, only one single species of an mRNA transcript corresponding in size to the cdh gene (2.5 kb) The relative amount of this transcript decreased as a function of time. Southern blot experiments done with genomic DNA and database search in the recently available genome information also ruled out the presence in this strain of a separate cbq gene distinct from the cdh gene. Taken together, these results demonstrated that CBQ originates from the cdh gene. Furthermore, it is not produced by differential splicing but by a posttranslational, predominantly intracellular, proteolytic cleavage.
ISSN:0167-4781
0006-3002
1879-2634
1879-2634
DOI:10.1016/S0167-4781(02)00243-9