Influence ofcis Element Arrangement on Promoter Strength in Trichoderma reesei

Applied and Environmental Microbiology
Daniel P KiesenhoferAstrid R Mach-Aigner

Abstract

Trichoderma reesei can produce up to 100 g/liter of extracellular proteins. The major and industrially relevant products are cellobiohydrolase I (CBHI) and the hemicellulase XYNI. The genes encoding both enzymes are transcriptionally activated by the regulatory protein Xyr1. The first 850 nucleotides of thecbh1promoter contain 14 Xyr1-binding sites (XBS), and 8 XBS are present in thexyn1promoter. Some of these XBS are arranged in tandem and others as inverted repeats. One suchciselement, an inverted repeat, plays a crucial role in the inducibility of thexyn1promoter. We investigated the impact of the properties of suchciselements by shuffling them by insertion, exchange, deletion, and rearrangement ofciselements in both thecbh1andxyn1promoter. A promoter-reporter assay using theAspergillus nigergoxAgene allowed us to measure changes in the promoter strength and inducibility. Most strikingly, we found that an inverted repeat of XBS causes an important increase incbh1promoter strength and allows induction by xylan or wheat straw. Furthermore, evidence is provided that the distances ofciselements to the transcription start site have important influence on promoter activity. Our results suggest that the arrangement and distances of...Continue Reading

References

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Feb 28, 2016·Enzyme and Microbial Technology·Tom Tao Huang, John M Wages

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Citations

Jun 4, 2018·Applied Microbiology and Biotechnology·Janina KlugeUlrich Kück
Aug 9, 2020·Fungal biology and biotechnology·Christian DerntlAstrid Mach-Aigner
Oct 27, 2018·Frontiers in Bioengineering and Biotechnology·Elisabeth FitzBernhard Seiboth
Jul 31, 2019·International Journal of Molecular Sciences·Xinxin XuWei Zhang
Oct 20, 2018·Frontiers in Bioengineering and Biotechnology·Leonardo Martins-SantanaRafael Silva-Rocha
Nov 9, 2018·Frontiers in Bioengineering and Biotechnology·Gustavo Pagotto BorinJuliana Velasco de Castro Oliveira

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Datasets Mentioned

BETA
KT176093
KY780444
KY780445
KY780443
KY780480

Methods Mentioned

BETA
PCR
electrophoresis
protein assay

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