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Gewählte Publikation:

Ma, S; Preims, M; Piumi, F; Kappel, L; Seiboth, B; Record, E; Kracher, D; Ludwig, R.
(2017): Molecular and catalytic properties of fungal extracellular cellobiose dehydrogenase produced in prokaryotic and eukaryotic expression systems
MICROB CELL FACT. 2017; 16: FullText FullText_BOKU

Background: Cellobiose dehydrogenase (CDH) is an extracellular enzyme produced by lignocellulolytic fungi. cdh gene expression is high in cellulose containing media, but relatively low CDH concentrations are found in the supernatant of fungal cultures due to strong binding to cellulose. Therefore, heterologous expression of CDH in Pichia pastoris was employed in the last 15 years, but the obtained enzymes were over glycosylated and had a reduced specific activity. Results: We compare the well-established CDH expression host P. pastoris with the less frequently used hosts Escherichia coli, Aspergillus niger, and Trichoderma reesei. The study evaluates the produced quantity and protein homogeneity of Corynascus thermophilus CDH in the culture supernatants, the purification, and finally compares the enzymes in regard to cofactor loading, glycosylation, catalytic constants and thermostability. Conclusions: Whereas E. coli could only express the catalytic dehydrogenase domain of CDH, all eukaryotic hosts could express full length CDH including the cytochrome domain. The CDH produced by T. reesei was most similar to the CDH originally isolated from the fungus C. thermophilus in regard to glycosylation, cofactor loading and catalytic constants. Under the tested experimental conditions the fungal expression hosts produce CDH of superior quality and uniformity compared to P. pastoris.
Autor/innen der BOKU Wien:
Kracher Daniel
Ludwig Roland
Ma Su
Preims Marita
BOKU Gendermonitor:

Find related publications in this database (Keywords)
Aspergillus niger
Cellobiose dehydrogenase
Cofactor loading
Escherichia coli
Heterologous expression
Pichia pastoris
Trichoderma reesei

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