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Backgrounds  The fungus  Trichoderma reesei  is an important workhorse for expression of homologous or heterologous genes, and the inducible  cbh1  promoter is generally used. However, constitutive expression is more preferable in some cases than inducible expression that leads to production of unwanted cellulase components. In this work, constitutive promoters of  T. reesei  were screened and successfully used for high level homologous expression of xylanase II.    Results  The transcriptional profiles of 13 key genes that participate in glucose metabolism in  T. reesei  were analyzed by quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR). The results indicated that the mRNA levels of  pdc  (encoding pyruvate decarboxylase) and  eno  (encoding enolase) genes were much higher than other genes under high glucose conditions. Recombinant  T. reesei  strains that homologously expressed xylanase II were constructed by using the promoters of the  pdc  and  eno  genes, and they respectively produced 9266 IU/ml and 8866 IU/ml of xylanase activities in the cultivation supernatant in a medium with high glucose concentration. The productivities of xylanase II were 1.61 g/L (with the  pdc  promoter) and 1.52 g/L (with the  eno  promoter), approximately accounted for 83% and 82% of the total protein secreted by  T. reesei , respectively.    Conclusions  This work demonstrates the screening of constitutive promoters by using RT-qPCR in  T. reesei , and has obtained the highest expression of recombinant xylanase II to date by using these promoters.

Achieving efficient protein expression in Trichoderma reesei by using strong constitutive promoters