Expression of fungal thermotolerant endo-1,4-β-glucanase in transgenic barley seeds during germination

Anna-Maria Nuutila, Anneli Ritala, Ron Skadsen, Leena Mannonen, Veli Kauppinen

Research output: Contribution to journalArticleScientificpeer-review

51 Citations (Scopus)

Abstract

The malting quality of two barley cultivars, Kymppi and Golden Promise, was modified to better meet the requirements of the brewing process. The egl1 gene, coding for fungal thermotolerant endo-1,4-β-glucanase (EGI, cellulase), was transferred to the cultivars using particle bombardment, and transgenic plants were regenerated on bialaphos selection. Integration of the egl1 gene was confirmed by Southern blot hybridization. The transgenic seeds were screened for the expression of the heterologous EGI. Under the high-pI α-amylase promoter, the egl1 gene was expressed during germination. The heterologous enzyme was thermotolerant at 65 °C for 2 h, thus being suitable for mashing conditions. The amount of heterologous EGI produced by the seeds (ca. 0.025% of soluble seed protein), has been shown to be sufficient to reduce wort viscosity by decreasing the soluble β-glucan content. A decrease in the soluble β-glucan content in the wort improves the filtration rate of beer.
Original languageEnglish
Pages (from-to)777-783
Number of pages7
JournalPlant Molecular Biology
Volume41
Issue number6
DOIs
Publication statusPublished - 1999
MoE publication typeA1 Journal article-refereed

Fingerprint

Hordeum
Germination
Seeds
wort (brewing)
seed germination
Glucans
barley
genetically modified organisms
glucans
Fungal Genes
seeds
malting quality
mashing
genes
Cellulase
Genetically Modified Plants
brewing
cultivars
Amylases
Southern Blotting

Keywords

  • cellulase
  • heterologous expression
  • Hordeum vulgare L.
  • particle bombardment
  • thermotolerant endo-1.4-beta-glucanase
  • transgenic barley

Cite this

@article{3885bb0de869476cbb61849b3e20fdf8,
title = "Expression of fungal thermotolerant endo-1,4-β-glucanase in transgenic barley seeds during germination",
abstract = "The malting quality of two barley cultivars, Kymppi and Golden Promise, was modified to better meet the requirements of the brewing process. The egl1 gene, coding for fungal thermotolerant endo-1,4-β-glucanase (EGI, cellulase), was transferred to the cultivars using particle bombardment, and transgenic plants were regenerated on bialaphos selection. Integration of the egl1 gene was confirmed by Southern blot hybridization. The transgenic seeds were screened for the expression of the heterologous EGI. Under the high-pI α-amylase promoter, the egl1 gene was expressed during germination. The heterologous enzyme was thermotolerant at 65 °C for 2 h, thus being suitable for mashing conditions. The amount of heterologous EGI produced by the seeds (ca. 0.025{\%} of soluble seed protein), has been shown to be sufficient to reduce wort viscosity by decreasing the soluble β-glucan content. A decrease in the soluble β-glucan content in the wort improves the filtration rate of beer.",
keywords = "cellulase, heterologous expression, Hordeum vulgare L., particle bombardment, thermotolerant endo-1.4-beta-glucanase, transgenic barley",
author = "Anna-Maria Nuutila and Anneli Ritala and Ron Skadsen and Leena Mannonen and Veli Kauppinen",
year = "1999",
doi = "10.1023/A:1006318206471",
language = "English",
volume = "41",
pages = "777--783",
journal = "Plant Molecular Biology",
issn = "0167-4412",
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Expression of fungal thermotolerant endo-1,4-β-glucanase in transgenic barley seeds during germination. / Nuutila, Anna-Maria; Ritala, Anneli; Skadsen, Ron; Mannonen, Leena; Kauppinen, Veli.

In: Plant Molecular Biology, Vol. 41, No. 6, 1999, p. 777-783.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Expression of fungal thermotolerant endo-1,4-β-glucanase in transgenic barley seeds during germination

AU - Nuutila, Anna-Maria

AU - Ritala, Anneli

AU - Skadsen, Ron

AU - Mannonen, Leena

AU - Kauppinen, Veli

PY - 1999

Y1 - 1999

N2 - The malting quality of two barley cultivars, Kymppi and Golden Promise, was modified to better meet the requirements of the brewing process. The egl1 gene, coding for fungal thermotolerant endo-1,4-β-glucanase (EGI, cellulase), was transferred to the cultivars using particle bombardment, and transgenic plants were regenerated on bialaphos selection. Integration of the egl1 gene was confirmed by Southern blot hybridization. The transgenic seeds were screened for the expression of the heterologous EGI. Under the high-pI α-amylase promoter, the egl1 gene was expressed during germination. The heterologous enzyme was thermotolerant at 65 °C for 2 h, thus being suitable for mashing conditions. The amount of heterologous EGI produced by the seeds (ca. 0.025% of soluble seed protein), has been shown to be sufficient to reduce wort viscosity by decreasing the soluble β-glucan content. A decrease in the soluble β-glucan content in the wort improves the filtration rate of beer.

AB - The malting quality of two barley cultivars, Kymppi and Golden Promise, was modified to better meet the requirements of the brewing process. The egl1 gene, coding for fungal thermotolerant endo-1,4-β-glucanase (EGI, cellulase), was transferred to the cultivars using particle bombardment, and transgenic plants were regenerated on bialaphos selection. Integration of the egl1 gene was confirmed by Southern blot hybridization. The transgenic seeds were screened for the expression of the heterologous EGI. Under the high-pI α-amylase promoter, the egl1 gene was expressed during germination. The heterologous enzyme was thermotolerant at 65 °C for 2 h, thus being suitable for mashing conditions. The amount of heterologous EGI produced by the seeds (ca. 0.025% of soluble seed protein), has been shown to be sufficient to reduce wort viscosity by decreasing the soluble β-glucan content. A decrease in the soluble β-glucan content in the wort improves the filtration rate of beer.

KW - cellulase

KW - heterologous expression

KW - Hordeum vulgare L.

KW - particle bombardment

KW - thermotolerant endo-1.4-beta-glucanase

KW - transgenic barley

U2 - 10.1023/A:1006318206471

DO - 10.1023/A:1006318206471

M3 - Article

VL - 41

SP - 777

EP - 783

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SN - 0167-4412

IS - 6

ER -