Screening of candidate regulators for cellulase and hemicellulase production in Trichoderma reesei and identification of a factor essential for cellulase production

Biotechnology for Biofuels

Tiina M Pakula¹  Markku Saloheimo¹  Merja Penttilä¹  Marika Vitikainen¹  Mikko Arvas¹  Nina Aro¹  Ann Westerholm-Parvinen¹  Mari J Valkonen¹  Mari Häkkinen¹ 

[1]VTT Technical Research Centre of Finland, PO Box 1000 Tietotie 2, Espoo FI-02044, VTT, Finland

关键词: Transcriptional profiling;    Transcription factors;    Hemicellulase;    Gene regulation;    Co-regulation;    Cellulase;    Carbohydrate active enzymes;   

Others  :  793842
DOI  :  10.1186/1754-6834-7-14

 received in 2013-07-05, accepted in 2014-01-14,  发布年份 2014

【 摘 要 】

Background

The soft rot ascomycetal fungus Trichoderma reesei is utilized for industrial production of secreted enzymes, especially lignocellulose degrading enzymes. T. reesei uses several different enzymes for the degradation of plant cell wall-derived material, including 9 characterized cellulases, 15 characterized hemicellulases and at least 42 genes predicted to encode cellulolytic or hemicellulolytic activities. Production of cellulases and hemicellulases is modulated by environmental and physiological conditions. Several regulators affecting the expression of cellulase and hemicellulase genes have been identified but more factors still unknown are believed to be present in the genome of T. reesei.

Results

We have used transcriptional profiling data from T. reesei cultures in which cellulase/hemicellulase production was induced by the addition of different lignocellulose-derived materials to identify putative novel regulators for cellulase and hemicellulase genes. Based on this induction data, supplemented with other published genome-wide data on different protein production conditions, 28 candidate regulatory genes were selected for further studies and they were overexpressed in T. reesei. Overexpression of seven genes led to at least 1.5-fold increased production of cellulase and/or xylanase activity in the modified strains as compared to the parental strain. Deletion of gene 77513, here designated as ace3, was found to be detrimental for cellulase production and for the expression of several cellulase genes studied. This deletion also significantly reduced xylanase activity and expression of xylan-degrading enzyme genes. Furthermore, our data revealed the presence of co-regulated chromosomal regions containing carbohydrate-active enzyme genes and candidate regulatory genes.

Conclusions

Transcriptional profiling results from glycoside hydrolase induction experiments combined with a previous study of specific protein production conditions was shown to be an effective method for finding novel candidate regulatory genes affecting the production of cellulases and hemicellulases. Recombinant strains with improved cellulase and/or xylanase production properties were constructed, and a gene essential for cellulase gene expression was found. In addition, more evidence was gained on the chromatin level regional regulation of carbohydrate-active enzyme gene expression.

【 授权许可】

   
2014 Häkkinen et al.; licensee BioMed Central Ltd.

【 图 表 】

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