Trichoderma reesei is one of the most frequently used production organisms for expression of plant cell wall degrading enzymes, but is also applied for production of diverse performance proteins in industry. Therefore optimization of expression of the target proteins under the strong cellulase promotors is crucial, but we are also testing alternative constitutive and inducible promotors and terminators for application in heterologous expression.
From our extensive transcriptome data resulting from growth under diverse conditions and with different mutants, we select suitable promotors and terminators, check for potential antisense transcripts or variable transcription start points and test them for expression efficiency in our functional genomics studies.
Figure 1. Schematic representation of a variable overexpression vector to be constructed by yeast recombination and/or joint PCR approaches.
In addition to overexpression we are also testing known regulators for the relevance of phosphorylation sites by introducing alleles with modified open reading frames bearing amino acid exchanges.
Schmoll M., Kotlowski, C., Seibel, C., Liebmann B., Kubicek, C. P. (2010) Recombinant production of an Aspergillus nidulans class I hydrophobin, (DewA) in Hypocrea jecorina (Trichoderma reesei) is promoter-dependent, Appl Microbiol Biotechnol, 88(1):95-103.