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

27 Citations (Scopus)

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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