The effect of specific growth rate on protein synthesis and secretion in the filamentous fungus Trichoderma reesei

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Abstract

Trichoderma reesei was cultivated in chemostat cultures on lactose-containing medium. The cultures were characterized for growth, consumption of the carbon source and protein production. Secreted proteins were produced most efficiently at low specific growth rates, 0.022-0.033 h-1, the highest specific rate of total protein production being 4.1 mg g-1 h-1 at the specific growth rate 0.031 h-1. At low specific growth rates, up to 29% of the proteins produced were extracellular, in comparison to only 6-8% at high specific growth rates, 0.045-0.066 h-1. To analyse protein synthesis and secretion in more detail, metabolic labelling of proteins was applied to analyse production of the major secreted protein, cellobiohydrolase I (CBHI, Cel7A). Intracellular and extracellular labelled CBHI was quantified and analysed for pl isoforms in two-dimensional gels, and the synthesis and secretion rates of the molecule were determined. Both the specific rates of CBHI synthesis and secretion were highest at low specific growth rates, the optimum being at 0.031 h-1. However, at low specific growth rates the secretion rate/synthesis rate ratio was significantly lower than that at high specific growth rates, indicating that at low growth rates the capacity of cells to transport the protein becomes limiting. In accordance with the high level of protein production and limitation in the secretory capacity, the transcript levels of the unfolded protein response (UPR) target genes pdi1 and bip1 as well as the gene encoding the UPR transcription factor hac1 were induced.

Original languageEnglish
Pages (from-to)135-143
Number of pages9
JournalMicrobiology
Volume151
Issue number1
DOIs
Publication statusPublished - 1 Jan 2005
MoE publication typeA1 Journal article-refereed

Fingerprint

Trichoderma
Fungi
Growth
Proteins
Unfolded Protein Response
Cellulose 1,4-beta-Cellobiosidase
Lactose
Genes
Carrier Proteins
Protein Isoforms
Transcription Factors
Carbon
Gels

Keywords

  • Unfolded-protein response (UPR)
  • Cellobiohydrolase I
  • Endoglucanase I
  • Dilution rate

Cite this

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title = "The effect of specific growth rate on protein synthesis and secretion in the filamentous fungus Trichoderma reesei",
abstract = "Trichoderma reesei was cultivated in chemostat cultures on lactose-containing medium. The cultures were characterized for growth, consumption of the carbon source and protein production. Secreted proteins were produced most efficiently at low specific growth rates, 0.022-0.033 h-1, the highest specific rate of total protein production being 4.1 mg g-1 h-1 at the specific growth rate 0.031 h-1. At low specific growth rates, up to 29{\%} of the proteins produced were extracellular, in comparison to only 6-8{\%} at high specific growth rates, 0.045-0.066 h-1. To analyse protein synthesis and secretion in more detail, metabolic labelling of proteins was applied to analyse production of the major secreted protein, cellobiohydrolase I (CBHI, Cel7A). Intracellular and extracellular labelled CBHI was quantified and analysed for pl isoforms in two-dimensional gels, and the synthesis and secretion rates of the molecule were determined. Both the specific rates of CBHI synthesis and secretion were highest at low specific growth rates, the optimum being at 0.031 h-1. However, at low specific growth rates the secretion rate/synthesis rate ratio was significantly lower than that at high specific growth rates, indicating that at low growth rates the capacity of cells to transport the protein becomes limiting. In accordance with the high level of protein production and limitation in the secretory capacity, the transcript levels of the unfolded protein response (UPR) target genes pdi1 and bip1 as well as the gene encoding the UPR transcription factor hac1 were induced.",
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The effect of specific growth rate on protein synthesis and secretion in the filamentous fungus Trichoderma reesei. / Pakula, Tiina M.; Salonen, Katri; Uusitalo, Jaana; Penttilä, Merja.

In: Microbiology, Vol. 151, No. 1, 01.01.2005, p. 135-143.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - The effect of specific growth rate on protein synthesis and secretion in the filamentous fungus Trichoderma reesei

AU - Pakula, Tiina M.

AU - Salonen, Katri

AU - Uusitalo, Jaana

AU - Penttilä, Merja

PY - 2005/1/1

Y1 - 2005/1/1

N2 - Trichoderma reesei was cultivated in chemostat cultures on lactose-containing medium. The cultures were characterized for growth, consumption of the carbon source and protein production. Secreted proteins were produced most efficiently at low specific growth rates, 0.022-0.033 h-1, the highest specific rate of total protein production being 4.1 mg g-1 h-1 at the specific growth rate 0.031 h-1. At low specific growth rates, up to 29% of the proteins produced were extracellular, in comparison to only 6-8% at high specific growth rates, 0.045-0.066 h-1. To analyse protein synthesis and secretion in more detail, metabolic labelling of proteins was applied to analyse production of the major secreted protein, cellobiohydrolase I (CBHI, Cel7A). Intracellular and extracellular labelled CBHI was quantified and analysed for pl isoforms in two-dimensional gels, and the synthesis and secretion rates of the molecule were determined. Both the specific rates of CBHI synthesis and secretion were highest at low specific growth rates, the optimum being at 0.031 h-1. However, at low specific growth rates the secretion rate/synthesis rate ratio was significantly lower than that at high specific growth rates, indicating that at low growth rates the capacity of cells to transport the protein becomes limiting. In accordance with the high level of protein production and limitation in the secretory capacity, the transcript levels of the unfolded protein response (UPR) target genes pdi1 and bip1 as well as the gene encoding the UPR transcription factor hac1 were induced.

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KW - Unfolded-protein response (UPR)

KW - Cellobiohydrolase I

KW - Endoglucanase I

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