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Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum

ß-1,6-glucanase produced by Trichoderma harzianum has been proven as one of the prime compounds to be excreted onto the hyphae of the pathogen causing localised cell wall lysis at the point of interaction. This study was conducted in the interest to investigate the regulation of ß-1,6-glucanase gene...

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Main Authors: Mustafa, Muskhazli, Wallis, G.L.F., Peberdy, J.F., Noor, Nurul Shafiza, Ahmad Bedawi, Salifah Hasanah, Go, Rusea, Abd Aziz, Nor Azwady
Format: Article
Language:English
English
Published: Universiti Kebangsaan Malaya 2009
Online Access:http://psasir.upm.edu.my/id/eprint/16394/1/Purification%20of%20enzyme%20and%20influence%20of%20substrate%20specificity%20on%20%C3%A2.pdf
http://psasir.upm.edu.my/id/eprint/16394/
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spelling my.upm.eprints.163942015-10-23T01:02:32Z http://psasir.upm.edu.my/id/eprint/16394/ Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum Mustafa, Muskhazli Wallis, G.L.F. Peberdy, J.F. Noor, Nurul Shafiza Ahmad Bedawi, Salifah Hasanah Go, Rusea Abd Aziz, Nor Azwady ß-1,6-glucanase produced by Trichoderma harzianum has been proven as one of the prime compounds to be excreted onto the hyphae of the pathogen causing localised cell wall lysis at the point of interaction. This study was conducted in the interest to investigate the regulation of ß-1,6-glucanase gene expression in T. harzianum strain BIO10671. β-1,6-glucanase enzyme from the culture filtrate of T. harzianum was purified through precipitation with 80% acetone followed by anion-exchange chromatography and chromatofocusing using Neobar AQ and Mono P HR 5/20 columns, respectively. Two β-1,6-glucanase bands at 32 kDa and 43 kDa in size has been purified. However, four restriction endonucleases digestion revealed only a single copy of ß-1,6-glucanase gene was encoded for both ß-1,6-glucanase isozymes. Fungal cell walls were able to trigger high level expression of gene encoding ß-1,6-glucanase. The expression of ß-1,6-glucanase gene was strongly affected by substrate specificity; where the presence of glucose or non ß-1,6-glucan linked substrate will significantly suppress the gene transcriptions. In spite of this, 24 hours were required for the gene transcription to achieve maximum total mRNA. Universiti Kebangsaan Malaya 2009 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/16394/1/Purification%20of%20enzyme%20and%20influence%20of%20substrate%20specificity%20on%20%C3%A2.pdf Mustafa, Muskhazli and Wallis, G.L.F. and Peberdy, J.F. and Noor, Nurul Shafiza and Ahmad Bedawi, Salifah Hasanah and Go, Rusea and Abd Aziz, Nor Azwady (2009) Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum. Sains Malaysiana, 38 (3). pp. 429-434. ISSN 0126-6039 English
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
English
description ß-1,6-glucanase produced by Trichoderma harzianum has been proven as one of the prime compounds to be excreted onto the hyphae of the pathogen causing localised cell wall lysis at the point of interaction. This study was conducted in the interest to investigate the regulation of ß-1,6-glucanase gene expression in T. harzianum strain BIO10671. β-1,6-glucanase enzyme from the culture filtrate of T. harzianum was purified through precipitation with 80% acetone followed by anion-exchange chromatography and chromatofocusing using Neobar AQ and Mono P HR 5/20 columns, respectively. Two β-1,6-glucanase bands at 32 kDa and 43 kDa in size has been purified. However, four restriction endonucleases digestion revealed only a single copy of ß-1,6-glucanase gene was encoded for both ß-1,6-glucanase isozymes. Fungal cell walls were able to trigger high level expression of gene encoding ß-1,6-glucanase. The expression of ß-1,6-glucanase gene was strongly affected by substrate specificity; where the presence of glucose or non ß-1,6-glucan linked substrate will significantly suppress the gene transcriptions. In spite of this, 24 hours were required for the gene transcription to achieve maximum total mRNA.
format Article
author Mustafa, Muskhazli
Wallis, G.L.F.
Peberdy, J.F.
Noor, Nurul Shafiza
Ahmad Bedawi, Salifah Hasanah
Go, Rusea
Abd Aziz, Nor Azwady
spellingShingle Mustafa, Muskhazli
Wallis, G.L.F.
Peberdy, J.F.
Noor, Nurul Shafiza
Ahmad Bedawi, Salifah Hasanah
Go, Rusea
Abd Aziz, Nor Azwady
Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum
author_facet Mustafa, Muskhazli
Wallis, G.L.F.
Peberdy, J.F.
Noor, Nurul Shafiza
Ahmad Bedawi, Salifah Hasanah
Go, Rusea
Abd Aziz, Nor Azwady
author_sort Mustafa, Muskhazli
title Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum
title_short Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum
title_full Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum
title_fullStr Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum
title_full_unstemmed Purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in Trichoderma harzianum
title_sort purification of enzyme and influence of substrate specificity on â-1,6-glucanase gene expression in trichoderma harzianum
publisher Universiti Kebangsaan Malaya
publishDate 2009
url http://psasir.upm.edu.my/id/eprint/16394/1/Purification%20of%20enzyme%20and%20influence%20of%20substrate%20specificity%20on%20%C3%A2.pdf
http://psasir.upm.edu.my/id/eprint/16394/
_version_ 1643826200919932928
score 13.211869

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