Release and acticity of bound alpha-amylase in a germinating barley grain

Tuomas Sopanen, Christiane Lauriere

Research output: Contribution to journalArticleScientificpeer-review

101 Citations (Scopus)

Abstract

In resting grains of Triumph barley (Hordeum vulgare L. cv Triumph) about 40% of the β-amylase could be extracted with a saline solution, the remaining 60% being in a bound form.
During seedling growth (20°C), the bound form was released mainly between days 1 and 3. When a preparation containing bound β-amylase was incubated with an extract made of endosperms separated from germinating grains, release of bound β-amylase took place and could be studied in vitro.
The release was almost completely prevented by leupeptin and antipain, specific inhibitors of a group of SH-proteinases, but it was not inhibited by pepstatin A or EDTA, which inhibit some other barley proteinases. It is thus very likely that in a whole grain, at least the bulk of the bound β-amylase is released by the proteolytic action of one or several SH-proteinases. When the bound β-amylase was released by papain, its molecular weight was about 5000 daltons smaller than that of β-amylase released by dithiothreitol. This indicates that the release is due to removal of a sequence of β-amylase itself.
A similar decrease in size took place during seedling growth. Bound β-amylase showed some activity against native starch and it hydrolyzed maltotetraose at a rate that was about 70% of the rate the same amount of bound β-amylase gave after release. Bound β-amylase is thus not inactive and it is likely that the slower rate of hydrolysis is due to steric hindrances which prevent substrates from reaching the active site.
Original languageEnglish
Pages (from-to)244-249
JournalPlant Physiology
Volume89
Issue number1
Publication statusPublished - 1989
MoE publication typeA1 Journal article-refereed

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alpha-Amylases
Hordeum
Amylases
alpha-amylase
amylases
barley
Peptide Hydrolases
proteinases
Seedlings
seedling growth
Antipain
Endosperm
Papain
dithiothreitol
papain
Dithiothreitol
whole grain foods
Growth
Sodium Chloride
active sites

Cite this

Sopanen, Tuomas ; Lauriere, Christiane. / Release and acticity of bound alpha-amylase in a germinating barley grain. In: Plant Physiology. 1989 ; Vol. 89, No. 1. pp. 244-249.
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abstract = "In resting grains of Triumph barley (Hordeum vulgare L. cv Triumph) about 40{\%} of the β-amylase could be extracted with a saline solution, the remaining 60{\%} being in a bound form. During seedling growth (20°C), the bound form was released mainly between days 1 and 3. When a preparation containing bound β-amylase was incubated with an extract made of endosperms separated from germinating grains, release of bound β-amylase took place and could be studied in vitro. The release was almost completely prevented by leupeptin and antipain, specific inhibitors of a group of SH-proteinases, but it was not inhibited by pepstatin A or EDTA, which inhibit some other barley proteinases. It is thus very likely that in a whole grain, at least the bulk of the bound β-amylase is released by the proteolytic action of one or several SH-proteinases. When the bound β-amylase was released by papain, its molecular weight was about 5000 daltons smaller than that of β-amylase released by dithiothreitol. This indicates that the release is due to removal of a sequence of β-amylase itself. A similar decrease in size took place during seedling growth. Bound β-amylase showed some activity against native starch and it hydrolyzed maltotetraose at a rate that was about 70{\%} of the rate the same amount of bound β-amylase gave after release. Bound β-amylase is thus not inactive and it is likely that the slower rate of hydrolysis is due to steric hindrances which prevent substrates from reaching the active site.",
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Sopanen, T & Lauriere, C 1989, 'Release and acticity of bound alpha-amylase in a germinating barley grain', Plant Physiology, vol. 89, no. 1, pp. 244-249.

Release and acticity of bound alpha-amylase in a germinating barley grain. / Sopanen, Tuomas; Lauriere, Christiane.

In: Plant Physiology, Vol. 89, No. 1, 1989, p. 244-249.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Release and acticity of bound alpha-amylase in a germinating barley grain

AU - Sopanen, Tuomas

AU - Lauriere, Christiane

PY - 1989

Y1 - 1989

N2 - In resting grains of Triumph barley (Hordeum vulgare L. cv Triumph) about 40% of the β-amylase could be extracted with a saline solution, the remaining 60% being in a bound form. During seedling growth (20°C), the bound form was released mainly between days 1 and 3. When a preparation containing bound β-amylase was incubated with an extract made of endosperms separated from germinating grains, release of bound β-amylase took place and could be studied in vitro. The release was almost completely prevented by leupeptin and antipain, specific inhibitors of a group of SH-proteinases, but it was not inhibited by pepstatin A or EDTA, which inhibit some other barley proteinases. It is thus very likely that in a whole grain, at least the bulk of the bound β-amylase is released by the proteolytic action of one or several SH-proteinases. When the bound β-amylase was released by papain, its molecular weight was about 5000 daltons smaller than that of β-amylase released by dithiothreitol. This indicates that the release is due to removal of a sequence of β-amylase itself. A similar decrease in size took place during seedling growth. Bound β-amylase showed some activity against native starch and it hydrolyzed maltotetraose at a rate that was about 70% of the rate the same amount of bound β-amylase gave after release. Bound β-amylase is thus not inactive and it is likely that the slower rate of hydrolysis is due to steric hindrances which prevent substrates from reaching the active site.

AB - In resting grains of Triumph barley (Hordeum vulgare L. cv Triumph) about 40% of the β-amylase could be extracted with a saline solution, the remaining 60% being in a bound form. During seedling growth (20°C), the bound form was released mainly between days 1 and 3. When a preparation containing bound β-amylase was incubated with an extract made of endosperms separated from germinating grains, release of bound β-amylase took place and could be studied in vitro. The release was almost completely prevented by leupeptin and antipain, specific inhibitors of a group of SH-proteinases, but it was not inhibited by pepstatin A or EDTA, which inhibit some other barley proteinases. It is thus very likely that in a whole grain, at least the bulk of the bound β-amylase is released by the proteolytic action of one or several SH-proteinases. When the bound β-amylase was released by papain, its molecular weight was about 5000 daltons smaller than that of β-amylase released by dithiothreitol. This indicates that the release is due to removal of a sequence of β-amylase itself. A similar decrease in size took place during seedling growth. Bound β-amylase showed some activity against native starch and it hydrolyzed maltotetraose at a rate that was about 70% of the rate the same amount of bound β-amylase gave after release. Bound β-amylase is thus not inactive and it is likely that the slower rate of hydrolysis is due to steric hindrances which prevent substrates from reaching the active site.

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