Transcriptional and proteomic changes in Trichoderma reesei expressing a heterologous bacterial xynB gene from the thermophile Dictyoglomus thermophilum

Helena Nevalainen, Johan Hekelaar, Jaana Uusitalo, Junior Te o, Iris Jonkers, Peter Bergquist, Merja Penttilä

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

We have taken a holistic approach into analysis of heterologous gene expression in Trichoderma involving enzyme activity measurements, gene transcription assays, zymogram gels, Western blotting and proteomics. The T. reesei transformant expressing the bacterial xynB gene encoding a thermophile xylanase was grown in a laboratory fermenter and assays were performed on culture supernatants collected from different time points. Transcription of the xynB under the T. reesei cbhl (cellobiohydrolase 1) promoter was comparable to that of the native cbh1, however, the yield of the heterologous xylanase produced in this particular transformant was no more than about 100 mg/l. Transcription analysis of selected genes involved in UPR (unfolded protein response) indicated a slight increase in the expression of pdi (protein disulfide isomerase) but otherwise the UPR pathway seemed not have been induced. Comparative proteomic analysis of the culture supernatant from the non-transformed host and the transformant expressing thermophilic xylanase showed differential expression of 23 proteins. Analysis of these spots is underway. This multifaceted approach may reveal crucial proteins involved in heterologous gene expression in filamentous fungi in general and Trichoderma in particular.
Original languageEnglish
Title of host publicationPoster Abstracts
Pages201
Publication statusPublished - 2004
Event7th European Conference on Fungal Genetics - Copenhagen, Denmark
Duration: 17 Apr 200420 Apr 2004

Conference

Conference7th European Conference on Fungal Genetics
CountryDenmark
CityCopenhagen
Period17/04/0420/04/04

Fingerprint

Dictyoglomus thermophilum
thermophilic microorganisms
Trichoderma reesei
xylanases
proteomics
unfolded protein response
transcription (genetics)
Trichoderma
protein disulfide-isomerase
cellulose 1,4-beta-cellobiosidase
genes
fermenters
assays
Western blotting
proteins
promoter regions
gels
enzyme activity
fungi

Cite this

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title = "Transcriptional and proteomic changes in Trichoderma reesei expressing a heterologous bacterial xynB gene from the thermophile Dictyoglomus thermophilum",
abstract = "We have taken a holistic approach into analysis of heterologous gene expression in Trichoderma involving enzyme activity measurements, gene transcription assays, zymogram gels, Western blotting and proteomics. The T. reesei transformant expressing the bacterial xynB gene encoding a thermophile xylanase was grown in a laboratory fermenter and assays were performed on culture supernatants collected from different time points. Transcription of the xynB under the T. reesei cbhl (cellobiohydrolase 1) promoter was comparable to that of the native cbh1, however, the yield of the heterologous xylanase produced in this particular transformant was no more than about 100 mg/l. Transcription analysis of selected genes involved in UPR (unfolded protein response) indicated a slight increase in the expression of pdi (protein disulfide isomerase) but otherwise the UPR pathway seemed not have been induced. Comparative proteomic analysis of the culture supernatant from the non-transformed host and the transformant expressing thermophilic xylanase showed differential expression of 23 proteins. Analysis of these spots is underway. This multifaceted approach may reveal crucial proteins involved in heterologous gene expression in filamentous fungi in general and Trichoderma in particular.",
author = "Helena Nevalainen and Johan Hekelaar and Jaana Uusitalo and {Te o}, Junior and Iris Jonkers and Peter Bergquist and Merja Penttil{\"a}",
year = "2004",
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Nevalainen, H, Hekelaar, J, Uusitalo, J, Te o, J, Jonkers, I, Bergquist, P & Penttilä, M 2004, Transcriptional and proteomic changes in Trichoderma reesei expressing a heterologous bacterial xynB gene from the thermophile Dictyoglomus thermophilum. in Poster Abstracts., VIIIp-30, pp. 201, 7th European Conference on Fungal Genetics, Copenhagen, Denmark, 17/04/04.

Transcriptional and proteomic changes in Trichoderma reesei expressing a heterologous bacterial xynB gene from the thermophile Dictyoglomus thermophilum. / Nevalainen, Helena; Hekelaar, Johan; Uusitalo, Jaana; Te o, Junior; Jonkers, Iris; Bergquist, Peter; Penttilä, Merja.

Poster Abstracts. 2004. p. 201 VIIIp-30.

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

TY - CHAP

T1 - Transcriptional and proteomic changes in Trichoderma reesei expressing a heterologous bacterial xynB gene from the thermophile Dictyoglomus thermophilum

AU - Nevalainen, Helena

AU - Hekelaar, Johan

AU - Uusitalo, Jaana

AU - Te o, Junior

AU - Jonkers, Iris

AU - Bergquist, Peter

AU - Penttilä, Merja

PY - 2004

Y1 - 2004

N2 - We have taken a holistic approach into analysis of heterologous gene expression in Trichoderma involving enzyme activity measurements, gene transcription assays, zymogram gels, Western blotting and proteomics. The T. reesei transformant expressing the bacterial xynB gene encoding a thermophile xylanase was grown in a laboratory fermenter and assays were performed on culture supernatants collected from different time points. Transcription of the xynB under the T. reesei cbhl (cellobiohydrolase 1) promoter was comparable to that of the native cbh1, however, the yield of the heterologous xylanase produced in this particular transformant was no more than about 100 mg/l. Transcription analysis of selected genes involved in UPR (unfolded protein response) indicated a slight increase in the expression of pdi (protein disulfide isomerase) but otherwise the UPR pathway seemed not have been induced. Comparative proteomic analysis of the culture supernatant from the non-transformed host and the transformant expressing thermophilic xylanase showed differential expression of 23 proteins. Analysis of these spots is underway. This multifaceted approach may reveal crucial proteins involved in heterologous gene expression in filamentous fungi in general and Trichoderma in particular.

AB - We have taken a holistic approach into analysis of heterologous gene expression in Trichoderma involving enzyme activity measurements, gene transcription assays, zymogram gels, Western blotting and proteomics. The T. reesei transformant expressing the bacterial xynB gene encoding a thermophile xylanase was grown in a laboratory fermenter and assays were performed on culture supernatants collected from different time points. Transcription of the xynB under the T. reesei cbhl (cellobiohydrolase 1) promoter was comparable to that of the native cbh1, however, the yield of the heterologous xylanase produced in this particular transformant was no more than about 100 mg/l. Transcription analysis of selected genes involved in UPR (unfolded protein response) indicated a slight increase in the expression of pdi (protein disulfide isomerase) but otherwise the UPR pathway seemed not have been induced. Comparative proteomic analysis of the culture supernatant from the non-transformed host and the transformant expressing thermophilic xylanase showed differential expression of 23 proteins. Analysis of these spots is underway. This multifaceted approach may reveal crucial proteins involved in heterologous gene expression in filamentous fungi in general and Trichoderma in particular.

M3 - Conference abstract in proceedings

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BT - Poster Abstracts

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