Secretion of a heat-stable fungal beta-glucanase from transgenic, suspension-cultured barley cells

Kristian Aspegren, Leena Mannonen, Anneli Ritala, Riitta Puupponen-Pimiä, Ulrika Kurten, Marjatta Salmenkallio-Marttila, Veli Kauppinen, Teemu Teeri

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

Transgenic plant cell cultures have a potential for production and secretion of important proteins and peptides. To assess the possibilities of using a stable barley suspension culture for secretion of heterologous proteins in active form, we expressed the cDNA of the thermostableβ-glucanase (EGI) ofTrichoderma reesei in barley suspension cells. The cDNA coding for EGI and its signal sequence was placed under the control of the CaMV 35S promoter and the construction was transferred to the cells by particle bombardment. Stably transformed lines were obtained by selecting for a cotransformed antibiotic resistance marker. The expression of EGI cDNA led to accumulation of EGI in the culture medium, as shown by analysis with EGI-specific antibodies. Enzymatic assays confirmed that the EGI secreted by the suspension cells retained its activity and thermostable character. Furthermore, it was shown that the enzyme produced by the transgenic suspension culture could be used for degradation of solubleβ-glucans during mashing.
Original languageEnglish
Pages (from-to)91-99
JournalMolecular Breeding
Volume1
Issue number1
DOIs
Publication statusPublished - 1995
MoE publication typeA1 Journal article-refereed

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beta-glucanase
Hordeum
cell suspension culture
Cultured Cells
Suspensions
Hot Temperature
barley
secretion
genetically modified organisms
heat
protein secretion
Complementary DNA
glucans
signal peptide
antibiotic resistance
transgenic plants
cell culture
culture media
promoter regions
peptides

Cite this

Aspegren, Kristian ; Mannonen, Leena ; Ritala, Anneli ; Puupponen-Pimiä, Riitta ; Kurten, Ulrika ; Salmenkallio-Marttila, Marjatta ; Kauppinen, Veli ; Teeri, Teemu. / Secretion of a heat-stable fungal beta-glucanase from transgenic, suspension-cultured barley cells. In: Molecular Breeding. 1995 ; Vol. 1, No. 1. pp. 91-99.
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title = "Secretion of a heat-stable fungal beta-glucanase from transgenic, suspension-cultured barley cells",
abstract = "Transgenic plant cell cultures have a potential for production and secretion of important proteins and peptides. To assess the possibilities of using a stable barley suspension culture for secretion of heterologous proteins in active form, we expressed the cDNA of the thermostableβ-glucanase (EGI) ofTrichoderma reesei in barley suspension cells. The cDNA coding for EGI and its signal sequence was placed under the control of the CaMV 35S promoter and the construction was transferred to the cells by particle bombardment. Stably transformed lines were obtained by selecting for a cotransformed antibiotic resistance marker. The expression of EGI cDNA led to accumulation of EGI in the culture medium, as shown by analysis with EGI-specific antibodies. Enzymatic assays confirmed that the EGI secreted by the suspension cells retained its activity and thermostable character. Furthermore, it was shown that the enzyme produced by the transgenic suspension culture could be used for degradation of solubleβ-glucans during mashing.",
author = "Kristian Aspegren and Leena Mannonen and Anneli Ritala and Riitta Puupponen-Pimi{\"a} and Ulrika Kurten and Marjatta Salmenkallio-Marttila and Veli Kauppinen and Teemu Teeri",
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Aspegren, K, Mannonen, L, Ritala, A, Puupponen-Pimiä, R, Kurten, U, Salmenkallio-Marttila, M, Kauppinen, V & Teeri, T 1995, 'Secretion of a heat-stable fungal beta-glucanase from transgenic, suspension-cultured barley cells', Molecular Breeding, vol. 1, no. 1, pp. 91-99. https://doi.org/10.1007/BF01682092

Secretion of a heat-stable fungal beta-glucanase from transgenic, suspension-cultured barley cells. / Aspegren, Kristian; Mannonen, Leena; Ritala, Anneli; Puupponen-Pimiä, Riitta; Kurten, Ulrika; Salmenkallio-Marttila, Marjatta; Kauppinen, Veli; Teeri, Teemu.

In: Molecular Breeding, Vol. 1, No. 1, 1995, p. 91-99.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Secretion of a heat-stable fungal beta-glucanase from transgenic, suspension-cultured barley cells

AU - Aspegren, Kristian

AU - Mannonen, Leena

AU - Ritala, Anneli

AU - Puupponen-Pimiä, Riitta

AU - Kurten, Ulrika

AU - Salmenkallio-Marttila, Marjatta

AU - Kauppinen, Veli

AU - Teeri, Teemu

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PY - 1995

Y1 - 1995

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AB - Transgenic plant cell cultures have a potential for production and secretion of important proteins and peptides. To assess the possibilities of using a stable barley suspension culture for secretion of heterologous proteins in active form, we expressed the cDNA of the thermostableβ-glucanase (EGI) ofTrichoderma reesei in barley suspension cells. The cDNA coding for EGI and its signal sequence was placed under the control of the CaMV 35S promoter and the construction was transferred to the cells by particle bombardment. Stably transformed lines were obtained by selecting for a cotransformed antibiotic resistance marker. The expression of EGI cDNA led to accumulation of EGI in the culture medium, as shown by analysis with EGI-specific antibodies. Enzymatic assays confirmed that the EGI secreted by the suspension cells retained its activity and thermostable character. Furthermore, it was shown that the enzyme produced by the transgenic suspension culture could be used for degradation of solubleβ-glucans during mashing.

U2 - 10.1007/BF01682092

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