Kinetics of Protein Secretion in Trichoderma Reesei

Tiina Pakula, Katri Salonen, Jaana Uusitalo, Markku Saloheimo, Robert J. Aarts, Merja Penttilä

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

Abstract

In order to obtain knowledge of the protein secretion process in filamentous fungi, and to study the rate-limiting steps in protein production, we have developed methodology for measuring the kinetics of protein production and secretion by metabolic labelling in Trichoderma. The synthesis of the 35S- labelled proteins and their transport into the growth medium was monitored by 2D gel electrophoresis and/or immunoprecipitation. The major cellulose of T.
reesei, cellobiohydrolase I (CBHI), was used as a model protein in the studies. To study the protein production kinetics in a controlled physiological state, a chemostat cultivation system was set up for the fungus. In the chemostat cultivations, the intracellular production rate of labelled CBHI and the accumulation rate of the protein into the culture medium was measured at different dilution rates, and the corresponding specific steady-state mRNA levels were determined. With this methodology we have been able to follow the time course of protein synthesis, glycosylation and transport into the medium,
and obtain information on the synthesis vs. secretion capacity of the fungus in relation to growth rate.
Original languageEnglish
Title of host publicationAbstracts from the 3rd ECFG
Subtitle of host publicationEuropean Congress on Fungal Genetics
PublisherNew Prairie Press
DOIs
Publication statusPublished - 1996
Event3rd European Fungal Conference on Fungal Genetics - Münster, Germany
Duration: 27 Mar 199630 Mar 1996

Publication series

NameFungal Genetics Reports
PublisherNew Prairie Press
Number34
Volume43

Conference

Conference3rd European Fungal Conference on Fungal Genetics
CountryGermany
CityMünster
Period27/03/9630/03/96

Fingerprint

Trichoderma reesei
protein secretion
kinetics
cellulose 1,4-beta-cellobiosidase
proteins
fungi
culture media
secretion
synthesis
Trichoderma
glycosylation
physiological state
gel electrophoresis
cellulose
protein synthesis
methodology

Cite this

Pakula, T., Salonen, K., Uusitalo, J., Saloheimo, M., Aarts, R. J., & Penttilä, M. (1996). Kinetics of Protein Secretion in Trichoderma Reesei. In Abstracts from the 3rd ECFG: European Congress on Fungal Genetics New Prairie Press. Fungal Genetics Reports, No. 34, Vol.. 43 https://doi.org/10.4148/1941-4765.1330
Pakula, Tiina ; Salonen, Katri ; Uusitalo, Jaana ; Saloheimo, Markku ; Aarts, Robert J. ; Penttilä, Merja. / Kinetics of Protein Secretion in Trichoderma Reesei. Abstracts from the 3rd ECFG: European Congress on Fungal Genetics. New Prairie Press, 1996. (Fungal Genetics Reports; No. 34, Vol. 43).
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abstract = "In order to obtain knowledge of the protein secretion process in filamentous fungi, and to study the rate-limiting steps in protein production, we have developed methodology for measuring the kinetics of protein production and secretion by metabolic labelling in Trichoderma. The synthesis of the 35S- labelled proteins and their transport into the growth medium was monitored by 2D gel electrophoresis and/or immunoprecipitation. The major cellulose of T.reesei, cellobiohydrolase I (CBHI), was used as a model protein in the studies. To study the protein production kinetics in a controlled physiological state, a chemostat cultivation system was set up for the fungus. In the chemostat cultivations, the intracellular production rate of labelled CBHI and the accumulation rate of the protein into the culture medium was measured at different dilution rates, and the corresponding specific steady-state mRNA levels were determined. With this methodology we have been able to follow the time course of protein synthesis, glycosylation and transport into the medium,and obtain information on the synthesis vs. secretion capacity of the fungus in relation to growth rate.",
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Pakula, T, Salonen, K, Uusitalo, J, Saloheimo, M, Aarts, RJ & Penttilä, M 1996, Kinetics of Protein Secretion in Trichoderma Reesei. in Abstracts from the 3rd ECFG: European Congress on Fungal Genetics. New Prairie Press, Fungal Genetics Reports, no. 34, vol. 43, 3rd European Fungal Conference on Fungal Genetics, Münster, Germany, 27/03/96. https://doi.org/10.4148/1941-4765.1330

Kinetics of Protein Secretion in Trichoderma Reesei. / Pakula, Tiina; Salonen, Katri; Uusitalo, Jaana; Saloheimo, Markku; Aarts, Robert J.; Penttilä, Merja.

Abstracts from the 3rd ECFG: European Congress on Fungal Genetics. New Prairie Press, 1996. (Fungal Genetics Reports; No. 34, Vol. 43).

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

TY - CHAP

T1 - Kinetics of Protein Secretion in Trichoderma Reesei

AU - Pakula, Tiina

AU - Salonen, Katri

AU - Uusitalo, Jaana

AU - Saloheimo, Markku

AU - Aarts, Robert J.

AU - Penttilä, Merja

PY - 1996

Y1 - 1996

N2 - In order to obtain knowledge of the protein secretion process in filamentous fungi, and to study the rate-limiting steps in protein production, we have developed methodology for measuring the kinetics of protein production and secretion by metabolic labelling in Trichoderma. The synthesis of the 35S- labelled proteins and their transport into the growth medium was monitored by 2D gel electrophoresis and/or immunoprecipitation. The major cellulose of T.reesei, cellobiohydrolase I (CBHI), was used as a model protein in the studies. To study the protein production kinetics in a controlled physiological state, a chemostat cultivation system was set up for the fungus. In the chemostat cultivations, the intracellular production rate of labelled CBHI and the accumulation rate of the protein into the culture medium was measured at different dilution rates, and the corresponding specific steady-state mRNA levels were determined. With this methodology we have been able to follow the time course of protein synthesis, glycosylation and transport into the medium,and obtain information on the synthesis vs. secretion capacity of the fungus in relation to growth rate.

AB - In order to obtain knowledge of the protein secretion process in filamentous fungi, and to study the rate-limiting steps in protein production, we have developed methodology for measuring the kinetics of protein production and secretion by metabolic labelling in Trichoderma. The synthesis of the 35S- labelled proteins and their transport into the growth medium was monitored by 2D gel electrophoresis and/or immunoprecipitation. The major cellulose of T.reesei, cellobiohydrolase I (CBHI), was used as a model protein in the studies. To study the protein production kinetics in a controlled physiological state, a chemostat cultivation system was set up for the fungus. In the chemostat cultivations, the intracellular production rate of labelled CBHI and the accumulation rate of the protein into the culture medium was measured at different dilution rates, and the corresponding specific steady-state mRNA levels were determined. With this methodology we have been able to follow the time course of protein synthesis, glycosylation and transport into the medium,and obtain information on the synthesis vs. secretion capacity of the fungus in relation to growth rate.

U2 - 10.4148/1941-4765.1330

DO - 10.4148/1941-4765.1330

M3 - Conference abstract in proceedings

T3 - Fungal Genetics Reports

BT - Abstracts from the 3rd ECFG

PB - New Prairie Press

ER -

Pakula T, Salonen K, Uusitalo J, Saloheimo M, Aarts RJ, Penttilä M. Kinetics of Protein Secretion in Trichoderma Reesei. In Abstracts from the 3rd ECFG: European Congress on Fungal Genetics. New Prairie Press. 1996. (Fungal Genetics Reports; No. 34, Vol. 43). https://doi.org/10.4148/1941-4765.1330