Characterization and kinetic analysis of a thermostable GH3 β-glucosidase from Penicillum brasilianum

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Krogh, K.B.R.M. (författare): Danmarks Tekniske Universitet,Technical University of Denmark,Novozymes A/S

Olsen, Carsten L. (författare)

Salomon Johansen, Katja, 1969 (författare)

Hojer-Pedersen, Jesper (författare): Danmarks Tekniske Universitet,Technical University of Denmark,Novozymes A/S

Börjesson, Johan (författare): Novozymes A/S,Lunds universitet,Lund University

Olsson, Lisbeth, 1963 (författare): Chalmers tekniska högskola,Chalmers University of Technology,Danmarks Tekniske Universitet,Technical University of Denmark

(creator_code:org_t)

2009-09-16
2010
Engelska.
Ingår i: Applied Microbiology and Biotechnology. - : Springer Science and Business Media LLC. - 1432-0614 .- 0175-7598. ; 86:86, s. 143-154

Relaterad länk:: https://research.cha...; visa fler...; https://doi.org/10.1...; visa färre...

Abstract Ämnesord

Stäng

A GH3 β-glucosidase (BGL) from Penicillum brasilianum was purified to homogeneity after cultivation on a cellulose and xylan rich medium. The BGL was identified in a genomic library, and it was successfully expressed in Aspergillus oryzae. The BGL had excellent stability at elevated temperatures with no loss in activity after 24 h of incubation at 60°C at pH 4-6, and the BGL was shown to have significantly higher stability at these conditions in comparison to Novozym 188 and to other fungal GH3 BGLs reported in the literature. The BGL had significant lower afinity for cellobiose compared with the artificial substrate para-nitrophenyl-β-D-glucopyranoside (pNP-Glc)and further, pronounced substrate inhibition using p-NP-Glc. Kinetic studies demonstrated the high importance of using cellbiose as substrate and glucose as inhibitor to describe the inhibition kinetics of BGL taking place during cellulose hydrolysis. A novel assay was developed to characterize this glucose inhibition on cellobiose hydrolosis. The assay uses labelled glucose-13C6 as inhibitor and subsequent mass spectrometry analysis to quantify the hydrolysis rates

Av författaren/redakt...: Krogh, K.B.R.M.; Harris, P.V.; Olsen, Carsten L ...; Salomon Johansen ...; Hojer-Pedersen, ...; Börjesson, Johan; visa fler...; Olsson, Lisbeth, ...; visa färre...

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