Incubation of barley kernel sections with purified cell wall degrading enzymes

Karin Autio, Leena Mannonen, Kirsti Pietilä, Maarit Koskinen, Matti Siika-aho, Matti Linko

Research output: Contribution to journalArticleScientificpeer-review

11 Citations (Scopus)

Abstract

The effects of purified Trichoderma reesei endo‐xylanase, XYN II and endoglucanases, EG I and EG II on the cell walls of embedded barley cross sections were studied. The cell walls were stained with Calcofluor which is specific for (1–3)(1–4)‐β‐D‐glucans (mixed‐linked β‐glucans) in histochemical applications. Whereas endosperm cell walls remained highly fluorescent after incubation with buffer, the aleurone cell walls were only weakly fluorescent. Both EG I and EG II were effective in degrading the outer endosperm cell walls, but had little or no effect on the inner endosperm and aleurone cell walls. Corresponding sections incubated with XYN II showed highly intensive fluorescence of aleurone cell walls suggesting that the surface of alaurone cell walls contains a high level of xylan which is removed by xylanase treatment exposing a layer rich in mixed linked β‐glucans. This indicates that arabinoxylans and β‐glucans in the aleurone cell walls of barley exist as separate, macroscopic layers. Xylanase did not have any Influence on the microstructure of endosperm cell walls which are rich in mixed linked β‐glucans. However, incubation with EG II after xylanase treatment caused the aleurone cell walls to become very thin.

Original languageEnglish
Pages (from-to)427-432
Number of pages6
JournalJournal of the Institute of Brewing
Volume102
Issue number6
DOIs
Publication statusPublished - 1996
MoE publication typeA1 Journal article-refereed

Fingerprint

Hordeum
Cell Wall
barley
cell walls
aleurone cells
Enzymes
enzymes
seeds
Endosperm
Glucans
glucans
endosperm
xylanases
endo-1,4-beta-xylanase
Endo-1,4-beta Xylanases
Cellulases
Xylans
Trichoderma reesei
Trichoderma
arabinoxylan

Cite this

Autio, Karin ; Mannonen, Leena ; Pietilä, Kirsti ; Koskinen, Maarit ; Siika-aho, Matti ; Linko, Matti. / Incubation of barley kernel sections with purified cell wall degrading enzymes. In: Journal of the Institute of Brewing. 1996 ; Vol. 102, No. 6. pp. 427-432.
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abstract = "The effects of purified Trichoderma reesei endo‐xylanase, XYN II and endoglucanases, EG I and EG II on the cell walls of embedded barley cross sections were studied. The cell walls were stained with Calcofluor which is specific for (1–3)(1–4)‐β‐D‐glucans (mixed‐linked β‐glucans) in histochemical applications. Whereas endosperm cell walls remained highly fluorescent after incubation with buffer, the aleurone cell walls were only weakly fluorescent. Both EG I and EG II were effective in degrading the outer endosperm cell walls, but had little or no effect on the inner endosperm and aleurone cell walls. Corresponding sections incubated with XYN II showed highly intensive fluorescence of aleurone cell walls suggesting that the surface of alaurone cell walls contains a high level of xylan which is removed by xylanase treatment exposing a layer rich in mixed linked β‐glucans. This indicates that arabinoxylans and β‐glucans in the aleurone cell walls of barley exist as separate, macroscopic layers. Xylanase did not have any Influence on the microstructure of endosperm cell walls which are rich in mixed linked β‐glucans. However, incubation with EG II after xylanase treatment caused the aleurone cell walls to become very thin.",
author = "Karin Autio and Leena Mannonen and Kirsti Pietil{\"a} and Maarit Koskinen and Matti Siika-aho and Matti Linko",
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Incubation of barley kernel sections with purified cell wall degrading enzymes. / Autio, Karin; Mannonen, Leena; Pietilä, Kirsti; Koskinen, Maarit; Siika-aho, Matti; Linko, Matti.

In: Journal of the Institute of Brewing, Vol. 102, No. 6, 1996, p. 427-432.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Autio, Karin

AU - Mannonen, Leena

AU - Pietilä, Kirsti

AU - Koskinen, Maarit

AU - Siika-aho, Matti

AU - Linko, Matti

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N2 - The effects of purified Trichoderma reesei endo‐xylanase, XYN II and endoglucanases, EG I and EG II on the cell walls of embedded barley cross sections were studied. The cell walls were stained with Calcofluor which is specific for (1–3)(1–4)‐β‐D‐glucans (mixed‐linked β‐glucans) in histochemical applications. Whereas endosperm cell walls remained highly fluorescent after incubation with buffer, the aleurone cell walls were only weakly fluorescent. Both EG I and EG II were effective in degrading the outer endosperm cell walls, but had little or no effect on the inner endosperm and aleurone cell walls. Corresponding sections incubated with XYN II showed highly intensive fluorescence of aleurone cell walls suggesting that the surface of alaurone cell walls contains a high level of xylan which is removed by xylanase treatment exposing a layer rich in mixed linked β‐glucans. This indicates that arabinoxylans and β‐glucans in the aleurone cell walls of barley exist as separate, macroscopic layers. Xylanase did not have any Influence on the microstructure of endosperm cell walls which are rich in mixed linked β‐glucans. However, incubation with EG II after xylanase treatment caused the aleurone cell walls to become very thin.

AB - The effects of purified Trichoderma reesei endo‐xylanase, XYN II and endoglucanases, EG I and EG II on the cell walls of embedded barley cross sections were studied. The cell walls were stained with Calcofluor which is specific for (1–3)(1–4)‐β‐D‐glucans (mixed‐linked β‐glucans) in histochemical applications. Whereas endosperm cell walls remained highly fluorescent after incubation with buffer, the aleurone cell walls were only weakly fluorescent. Both EG I and EG II were effective in degrading the outer endosperm cell walls, but had little or no effect on the inner endosperm and aleurone cell walls. Corresponding sections incubated with XYN II showed highly intensive fluorescence of aleurone cell walls suggesting that the surface of alaurone cell walls contains a high level of xylan which is removed by xylanase treatment exposing a layer rich in mixed linked β‐glucans. This indicates that arabinoxylans and β‐glucans in the aleurone cell walls of barley exist as separate, macroscopic layers. Xylanase did not have any Influence on the microstructure of endosperm cell walls which are rich in mixed linked β‐glucans. However, incubation with EG II after xylanase treatment caused the aleurone cell walls to become very thin.

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