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The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant

BACKGROUND: The filamentous fungus Trichoderma reesei (anamorph of Hypocrea jecorina) produces increased cellulase expression when grown on cellulose or its derivatives as a sole carbon source. It has been believed that β-glucosidases of T. reesei not only metabolize cellobiose but also contribute i...

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Autores principales: Shida, Yosuke, Yamaguchi, Kaori, Nitta, Mikiko, Nakamura, Ayana, Takahashi, Machiko, Kidokoro, Shun-ichi, Mori, Kazuki, Tashiro, Kosuke, Kuhara, Satoru, Matsuzawa, Tomohiko, Yaoi, Katsuro, Sakamoto, Yasumitsu, Tanaka, Nobutada, Morikawa, Yasushi, Ogasawara, Wataru
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4696228/
https://www.ncbi.nlm.nih.gov/pubmed/26719764
http://dx.doi.org/10.1186/s13068-015-0420-y
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author Shida, Yosuke
Yamaguchi, Kaori
Nitta, Mikiko
Nakamura, Ayana
Takahashi, Machiko
Kidokoro, Shun-ichi
Mori, Kazuki
Tashiro, Kosuke
Kuhara, Satoru
Matsuzawa, Tomohiko
Yaoi, Katsuro
Sakamoto, Yasumitsu
Tanaka, Nobutada
Morikawa, Yasushi
Ogasawara, Wataru
author_facet Shida, Yosuke
Yamaguchi, Kaori
Nitta, Mikiko
Nakamura, Ayana
Takahashi, Machiko
Kidokoro, Shun-ichi
Mori, Kazuki
Tashiro, Kosuke
Kuhara, Satoru
Matsuzawa, Tomohiko
Yaoi, Katsuro
Sakamoto, Yasumitsu
Tanaka, Nobutada
Morikawa, Yasushi
Ogasawara, Wataru
author_sort Shida, Yosuke
collection PubMed
description BACKGROUND: The filamentous fungus Trichoderma reesei (anamorph of Hypocrea jecorina) produces increased cellulase expression when grown on cellulose or its derivatives as a sole carbon source. It has been believed that β-glucosidases of T. reesei not only metabolize cellobiose but also contribute in the production of inducers of cellulase gene expression by their transglycosylation activity. The cellulase hyper-producing mutant PC-3-7 developed in Japan has enhanced cellulase production ability when cellobiose is used as the inducer. The comparative genomics analysis of PC-3-7 and its parent revealed a single-nucleotide mutation within the bgl2 gene encoding intracellular β-glucosidase II (BGLII/Cel1a), giving rise to an amino acid substitution in PC-3-7, which could potentially account for the enhanced cellulase expression when these strains are cultivated on cellulose and cellobiose. RESULTS: To analyze the effects of the BGLII mutation in cellulase induction, we constructed both a bgl2 revertant and a disruptant. Enzymatic analysis of the transformant lysates showed that the strain expressing mutant BGLII exhibited weakened cellobiose hydrolytic activity, but produced some transglycosylation products, suggesting that the SNP in bgl2 strongly diminished cellobiase activity, but did not result in complete loss of function of BGLII. The analysis of the recombinant BGLII revealed that transglycosylation products might be oligosaccharides, composed probably of glucose linked β-1,4, β-1,3, or a mixture of both. PC-3-7 revertants of bgl2 exhibited reduced expression and inducibility of cellulase during growth on cellulose and cellobiose substrates. Furthermore, the effect of this bgl2 mutation was reproduced in the common strain QM9414 in which the transformants showed cellulase production comparable to that of PC-3-7. CONCLUSION: We conclude that BGLII plays an important role in cellulase induction in T. reesei and that the bgl2 mutation in PC-3-7 brought about enhanced cellulase expression on cellobiose. The results of the investigation using PC-3-7 suggested that other mutation(s) in PC-3-7 could also contribute to cellulase induction. Further investigation is essential to unravel the mechanism responsible for cellulase induction in T. reesei. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-015-0420-y) contains supplementary material, which is available to authorized users.
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spelling pubmed-46962282015-12-31 The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant Shida, Yosuke Yamaguchi, Kaori Nitta, Mikiko Nakamura, Ayana Takahashi, Machiko Kidokoro, Shun-ichi Mori, Kazuki Tashiro, Kosuke Kuhara, Satoru Matsuzawa, Tomohiko Yaoi, Katsuro Sakamoto, Yasumitsu Tanaka, Nobutada Morikawa, Yasushi Ogasawara, Wataru Biotechnol Biofuels Research BACKGROUND: The filamentous fungus Trichoderma reesei (anamorph of Hypocrea jecorina) produces increased cellulase expression when grown on cellulose or its derivatives as a sole carbon source. It has been believed that β-glucosidases of T. reesei not only metabolize cellobiose but also contribute in the production of inducers of cellulase gene expression by their transglycosylation activity. The cellulase hyper-producing mutant PC-3-7 developed in Japan has enhanced cellulase production ability when cellobiose is used as the inducer. The comparative genomics analysis of PC-3-7 and its parent revealed a single-nucleotide mutation within the bgl2 gene encoding intracellular β-glucosidase II (BGLII/Cel1a), giving rise to an amino acid substitution in PC-3-7, which could potentially account for the enhanced cellulase expression when these strains are cultivated on cellulose and cellobiose. RESULTS: To analyze the effects of the BGLII mutation in cellulase induction, we constructed both a bgl2 revertant and a disruptant. Enzymatic analysis of the transformant lysates showed that the strain expressing mutant BGLII exhibited weakened cellobiose hydrolytic activity, but produced some transglycosylation products, suggesting that the SNP in bgl2 strongly diminished cellobiase activity, but did not result in complete loss of function of BGLII. The analysis of the recombinant BGLII revealed that transglycosylation products might be oligosaccharides, composed probably of glucose linked β-1,4, β-1,3, or a mixture of both. PC-3-7 revertants of bgl2 exhibited reduced expression and inducibility of cellulase during growth on cellulose and cellobiose substrates. Furthermore, the effect of this bgl2 mutation was reproduced in the common strain QM9414 in which the transformants showed cellulase production comparable to that of PC-3-7. CONCLUSION: We conclude that BGLII plays an important role in cellulase induction in T. reesei and that the bgl2 mutation in PC-3-7 brought about enhanced cellulase expression on cellobiose. The results of the investigation using PC-3-7 suggested that other mutation(s) in PC-3-7 could also contribute to cellulase induction. Further investigation is essential to unravel the mechanism responsible for cellulase induction in T. reesei. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-015-0420-y) contains supplementary material, which is available to authorized users. BioMed Central 2015-12-30 /pmc/articles/PMC4696228/ /pubmed/26719764 http://dx.doi.org/10.1186/s13068-015-0420-y Text en © Shida et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Shida, Yosuke
Yamaguchi, Kaori
Nitta, Mikiko
Nakamura, Ayana
Takahashi, Machiko
Kidokoro, Shun-ichi
Mori, Kazuki
Tashiro, Kosuke
Kuhara, Satoru
Matsuzawa, Tomohiko
Yaoi, Katsuro
Sakamoto, Yasumitsu
Tanaka, Nobutada
Morikawa, Yasushi
Ogasawara, Wataru
The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant
title The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant
title_full The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant
title_fullStr The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant
title_full_unstemmed The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant
title_short The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant
title_sort impact of a single-nucleotide mutation of bgl2 on cellulase induction in a trichoderma reesei mutant
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4696228/
https://www.ncbi.nlm.nih.gov/pubmed/26719764
http://dx.doi.org/10.1186/s13068-015-0420-y
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