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 the cbh1 promoter contain 14 Xyr1-binding sites (XBS), and 8 XBS are present in the xyn1 promoter. Some of these XBS are arranged in tandem and others as inverted repeats. One such cis element, an inverted repeat, plays a crucial role in the inducibility of the xyn1 promoter. We investigated the impact of the properties of such cis elements by shuffling them by insertion, exchange, deletion, and rearrangement of cis elements in both the cbh1 and xyn1 promoter. A promoter-reporter assay using the Aspergillus nigergoxA gene 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 in cbh1 promoter strength and allows induction by xylan or wheat straw. Furthermore, evidence is provided that the distances of cis elements to the transcription start site have important influence on promoter activity. Our results suggest that the arrangement and distances of cis elements have large impacts on the strength of the cbh1 promoter, whereas the sheer number of XBS has only secondary importance. Ultimately, the biotechnologically important cbh1 promoter can be improved by cis element rearrangement.IMPORTANCE In the present study, we demonstrate that the arrangement of cis elements has a major impact on promoter strength and inducibility. We discovered an influence on promoter activity by the distances of cis elements to the transcription start site. Furthermore, we found that the configuration of cis elements has a greater effect on promoter strength than does the sheer number of transactivator binding sites present in the promoter. Altogether, the arrangement of cis elements is an important factor that should not be overlooked when enhancement of gene expression is desired.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Kiesenhofer DP,Mach RL,Mach-Aigner ARdoi
10.1128/AEM.01742-17subject
Has Abstractpub_date
2017-12-15 00:00:00issue
1eissn
0099-2240issn
1098-5336pii
AEM.01742-17journal_volume
84pub_type
杂志文章abstract::Pigment production by the fungus Monascus sp. was studied to determine why Monascus sp. provides more pigment in solid culture than in submerged culture. Adding a sterilized nonionic polymeric adsorbent resin directly to the growing submerged culture did not enhance the pigment production, thus indicating that pigment...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.47.6.1323-1326.1984
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journal_title:Applied and environmental microbiology
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doi:10.1128/AEM.44.2.355-362.1982
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journal_title:Applied and environmental microbiology
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doi:10.1128/AEM.32.6.741-746.1976
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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journal_title:Applied and environmental microbiology
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doi:10.1128/AEM.61.2.518-525.1995
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doi:10.1128/AEM.00531-07
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pub_type: 杂志文章
doi:10.1128/aem.69.10.6121-6127.2003
更新日期:2003-10-01 00:00:00
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pub_type: 杂志文章
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.39.5.1078-1080.1980
更新日期:1980-05-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.02037-13
更新日期:2014-04-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.42.3.427-432.1981
更新日期:1981-09-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.45.3.1066-1084.1983
更新日期:1983-03-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/AEM.45.4.1277-1282.1983
更新日期:1983-04-01 00:00:00
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pub_type: 杂志文章
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journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.57.9.2576-2580.1991
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/aem.70.2.822-830.2004
更新日期:2004-02-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2020-05-19 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.61.5.1938-1945.1995
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.67.8.3702-3706.2001
更新日期:2001-08-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.71.1.530-537.2005
更新日期:2005-01-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.54.8.1983-1988.1988
更新日期:1988-08-01 00:00:00