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Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis

BACKGROUND: Biologically relevant molecular markers can be used in evaluation of the physiological state of an organism in biotechnical processes. We monitored at high frequency the expression of 34 marker genes in batch, fed-batch and continuous cultures of the filamentous fungus Trichoderma reesei...

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Autores principales: Rautio, Jari J, Bailey, Michael, Kivioja, Teemu, Söderlund, Hans, Penttilä, Merja, Saloheimo, Markku
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1899492/
https://www.ncbi.nlm.nih.gov/pubmed/17537269
http://dx.doi.org/10.1186/1472-6750-7-28
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author Rautio, Jari J
Bailey, Michael
Kivioja, Teemu
Söderlund, Hans
Penttilä, Merja
Saloheimo, Markku
author_facet Rautio, Jari J
Bailey, Michael
Kivioja, Teemu
Söderlund, Hans
Penttilä, Merja
Saloheimo, Markku
author_sort Rautio, Jari J
collection PubMed
description BACKGROUND: Biologically relevant molecular markers can be used in evaluation of the physiological state of an organism in biotechnical processes. We monitored at high frequency the expression of 34 marker genes in batch, fed-batch and continuous cultures of the filamentous fungus Trichoderma reesei by the transcriptional analysis method TRAC (TRanscript analysis with the aid of Affinity Capture). Expression of specific genes was normalised either with respect to biomass or to overall polyA RNA concentration. Expressional variation of the genes involved in various process relevant cellular functions, such as protein production, growth and stress responses, was related to process parameters such as specific growth and production rates and substrate and dissolved oxygen concentrations. RESULTS: Gene expression of secreted cellulases and recombinant Melanocarpus albomyces laccase predicted the trends in the corresponding extracellular enzyme production rates and was highest in a narrow "physiological window" in the specific growth rate (μ) range of 0.03 – 0.05 h(-1). Expression of ribosomal protein mRNAs was consistent with the changes in μ. Nine starvation-related genes were found as potential markers for detection of insufficient substrate feed for maintaining optimal protein production. For two genes induced in anaerobic conditions, increasing transcript levels were measured as dissolved oxygen decreased. CONCLUSION: The data obtained by TRAC supported the usefulness of focused and intensive transcriptional analysis in monitoring of biotechnical processes providing thus tools for process optimisation purposes.
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spelling pubmed-18994922007-06-27 Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis Rautio, Jari J Bailey, Michael Kivioja, Teemu Söderlund, Hans Penttilä, Merja Saloheimo, Markku BMC Biotechnol Research Article BACKGROUND: Biologically relevant molecular markers can be used in evaluation of the physiological state of an organism in biotechnical processes. We monitored at high frequency the expression of 34 marker genes in batch, fed-batch and continuous cultures of the filamentous fungus Trichoderma reesei by the transcriptional analysis method TRAC (TRanscript analysis with the aid of Affinity Capture). Expression of specific genes was normalised either with respect to biomass or to overall polyA RNA concentration. Expressional variation of the genes involved in various process relevant cellular functions, such as protein production, growth and stress responses, was related to process parameters such as specific growth and production rates and substrate and dissolved oxygen concentrations. RESULTS: Gene expression of secreted cellulases and recombinant Melanocarpus albomyces laccase predicted the trends in the corresponding extracellular enzyme production rates and was highest in a narrow "physiological window" in the specific growth rate (μ) range of 0.03 – 0.05 h(-1). Expression of ribosomal protein mRNAs was consistent with the changes in μ. Nine starvation-related genes were found as potential markers for detection of insufficient substrate feed for maintaining optimal protein production. For two genes induced in anaerobic conditions, increasing transcript levels were measured as dissolved oxygen decreased. CONCLUSION: The data obtained by TRAC supported the usefulness of focused and intensive transcriptional analysis in monitoring of biotechnical processes providing thus tools for process optimisation purposes. BioMed Central 2007-05-30 /pmc/articles/PMC1899492/ /pubmed/17537269 http://dx.doi.org/10.1186/1472-6750-7-28 Text en Copyright © 2007 Rautio et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Rautio, Jari J
Bailey, Michael
Kivioja, Teemu
Söderlund, Hans
Penttilä, Merja
Saloheimo, Markku
Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis
title Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis
title_full Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis
title_fullStr Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis
title_full_unstemmed Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis
title_short Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis
title_sort physiological evaluation of the filamentous fungus trichoderma reesei in production processes by marker gene expression analysis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1899492/
https://www.ncbi.nlm.nih.gov/pubmed/17537269
http://dx.doi.org/10.1186/1472-6750-7-28
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