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Applied and Environmental Microbiology, September 2002, p. 4546-4553, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4546-4553.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Enzymatic Properties and Intracellular Localization of the Novel Trichoderma reesei ß-Glucosidase BGLII (Cel1A)

Markku Saloheimo,1* Juha Kuja-Panula,1,{dagger} Erkko Ylösmäki,1 Michael Ward,2 and Merja Penttilä1

VTT Biotechnology, 02044 VTT, Finland,1 Genencor International, Palo Alto, California 94304-10132

Received 11 March 2002/ Accepted 18 June 2002

This paper describes the characterization of an intracellular ß-glucosidase enzyme BGLII (Cel1a) and its gene (bgl2) from the cellulolytic fungus Trichoderma reesei (Hypocrea jecorina). The expression pattern of bgl2 is similar to that of other cellulase genes known from this fungus, and the gene would appear to be under the control of carbon catabolite repression mediated by the cre1 gene. The BGLII protein was produced in Escherichia coli, and its enzymatic properties were analyzed. It was shown to be a specific ß-glucosidase, having no ß-galactosidase side activity. It hydrolyzed both cellotriose and cellotetraose. BGLII exhibited transglycosylation activity, producing mainly cellotriose from cellobiose and sophorose and cellobiose from glucose. Antibodies raised against BGLII showed the presence of the enzyme in T. reesei cell lysates but not in the culture supernatant. Activity measurements and Western blot analysis of T. reesei strains expressing bgl2 from a constitutive promoter further confirmed the intracellular localization of this ß-glucosidase.

* Corresponding author. Mailing address: VTT Biotechnology, P.O. Box 1500, 02044 VTT, Finland. Phone: 358-9-456 78628. Fax: 358-9-455 2103. E-mail: Markku.Saloheimo{at}vtt.fi.

{dagger} Present address: Institute of Biotechnology, University of Helsinki, Helsinki, Finland.

Applied and Environmental Microbiology, September 2002, p. 4546-4553, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4546-4553.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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