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 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.
Raíces
Manuel
M
Division of Industrial Biotechnology, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Cubanacan, Havana, Cuba.
Montesino
Raquel
R
Cremata
José
J
García
Bianca
B
Perdomo
Walmer
W
Szabó
István
I
Henriksson
Gunnar
G
Hallberg
B Martin
BM
Pettersson
Göran
G
Johansson
Gunnar
G
D000483:Alleles; D015139:Blotting, Southern; D015153:Blotting, Western; D002237:Carbohydrate Dehydrogenases / Q000096:biosynthesis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry; D020075:Phanerochaete / Q000235:genetics / Q000378:metabolism*; D012697:Serine Endopeptidases; D014158:Transcription, Genetic
D005656:Fungal Proteins; D002237:Carbohydrate Dehydrogenases; C019859:cellobiose-quinone oxidoreductase; D012697:Serine Endopeptidases; C036813:lysyl endopeptidase