Expression and Secretion of Barley Cysteine Endopeptidase B and Cellobiohydrolase I in Trichoderma reesei.

Applied and Environmental Microbiology
Marko J NykänenAnita Mikkonen

Abstract

Localization of expression and secretion of a heterologous barley cysteine endopeptidase (EPB) and the homologous main cellobiohydrolase I (CBHI) in a Trichoderma reesei transformant expressing both proteins were studied. The transformant was grown on solid medium with Avicel cellulose and lactose to induce the cbh1 promoter for the synthesis of the native CBHI and the recombinant barley protein linked to a cbh1 expression cassette. Differences in localization of expression between the two proteins were clearly indicated by in situ hybridization, indirect immunofluorescence, and immunoelectron microscopy. In young hyphae, native-size recombinant epb mRNA was localized to apical compartments. In older cultures, it was also seen in subapical compartments but not in hyphae from the colony center. The recombinant EPB had a higher molecular weight than the native barley protein, probably due to glycosylation and differential processing in the fungal host. As was found with its transcripts, recombinant EPB was localized in apical and subapical compartments of hyphae. The cbh1 mRNA and CBHI were both localized to all hyphae of a colony, which suggests that the endogenous CBHI was also secreted from these. In immunoelectron microscopy,...Continue Reading

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Citations

Jun 8, 2007·The Journal of Biological Chemistry·Mari ValkonenRory R Duncan
Jul 9, 2014·Annual Review of Phytopathology·Christian P KubicekN Louise Glass
Nov 10, 2001·Annual Review of Phytopathology·Joan M HensonAlan W Day
Apr 18, 2002·Canadian Journal of Microbiology·Marko J NykänenAnita Mikkonen
Oct 15, 2011·Microbiology·Robyn Peterson, Helena Nevalainen
Feb 19, 2021·Journal of Industrial Microbiology & Biotechnology·Elizabeth BodieTim Dodge
Mar 15, 2006·Applied and Environmental Microbiology·R SaarelainenP Suominen
Sep 6, 2015·New Biotechnology·Angela SunHelena Nevalainen

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