Novel genetic tools that enable highly pure protein production in Trichoderma reesei
Trichoderma reesei is an established protein production host with high natural capacity to secrete enzymes. The lack of efficient genome engineering approaches and absence of robust constitutive gene expression systems limits exploitation of this organism in some protein production applications. Her...
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ftvttcrispub:oai:cris.vtt.fi:publications/90d0ad99-2abc-4248-916b-bea97ec91d88 2024-06-02T07:57:51+00:00 Novel genetic tools that enable highly pure protein production in Trichoderma reesei Rantasalo, Anssi Vitikainen, Marika Paasikallio, Toni Jäntti, Jussi Landowski, Christopher P. Mojzita, Dominik 2019-03-22 https://cris.vtt.fi/en/publications/90d0ad99-2abc-4248-916b-bea97ec91d88 https://doi.org/10.1038/s41598-019-41573-8 http://www.scopus.com/inward/record.url?scp=85063346796&partnerID=8YFLogxK eng eng https://cris.vtt.fi/en/publications/90d0ad99-2abc-4248-916b-bea97ec91d88 info:eu-repo/semantics/openAccess Rantasalo , A , Vitikainen , M , Paasikallio , T , Jäntti , J , Landowski , C P & Mojzita , D 2019 , ' Novel genetic tools that enable highly pure protein production in Trichoderma reesei ' , Scientific Reports , vol. 9 , no. 1 , 5032 . https://doi.org/10.1038/s41598-019-41573-8 article 2019 ftvttcrispub https://doi.org/10.1038/s41598-019-41573-8 2024-05-07T03:08:57Z Trichoderma reesei is an established protein production host with high natural capacity to secrete enzymes. The lack of efficient genome engineering approaches and absence of robust constitutive gene expression systems limits exploitation of this organism in some protein production applications. Here we report engineering of T. reesei for high-level production of highly enriched lipase B of Candida antarctica (calB) using glucose as a carbon source. Multiplexed CRISPR/Cas9 in combination with the use of our recently established synthetic expression system (SES) enabled accelerated construction of strains, which produced high amounts of highly pure calB. Using SES, calB production levels in cellulase-inducing medium were comparable to the levels obtained by using the commonly employed inducible cbh1 promoter, where a wide spectrum of native enzymes were co-produced. Due to highly constitutive expression provided by the SES, it was possible to carry out the production in cellulase-repressing glucose medium leading to around 4 grams per liter of fully functional calB and simultaneous elimination of unwanted background enzymes. Article in Journal/Newspaper Antarc* Antarctica VTT's Research Information Portal Scientific Reports 9 1 |
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VTT's Research Information Portal |
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ftvttcrispub |
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English |
description |
Trichoderma reesei is an established protein production host with high natural capacity to secrete enzymes. The lack of efficient genome engineering approaches and absence of robust constitutive gene expression systems limits exploitation of this organism in some protein production applications. Here we report engineering of T. reesei for high-level production of highly enriched lipase B of Candida antarctica (calB) using glucose as a carbon source. Multiplexed CRISPR/Cas9 in combination with the use of our recently established synthetic expression system (SES) enabled accelerated construction of strains, which produced high amounts of highly pure calB. Using SES, calB production levels in cellulase-inducing medium were comparable to the levels obtained by using the commonly employed inducible cbh1 promoter, where a wide spectrum of native enzymes were co-produced. Due to highly constitutive expression provided by the SES, it was possible to carry out the production in cellulase-repressing glucose medium leading to around 4 grams per liter of fully functional calB and simultaneous elimination of unwanted background enzymes. |
format |
Article in Journal/Newspaper |
author |
Rantasalo, Anssi Vitikainen, Marika Paasikallio, Toni Jäntti, Jussi Landowski, Christopher P. Mojzita, Dominik |
spellingShingle |
Rantasalo, Anssi Vitikainen, Marika Paasikallio, Toni Jäntti, Jussi Landowski, Christopher P. Mojzita, Dominik Novel genetic tools that enable highly pure protein production in Trichoderma reesei |
author_facet |
Rantasalo, Anssi Vitikainen, Marika Paasikallio, Toni Jäntti, Jussi Landowski, Christopher P. Mojzita, Dominik |
author_sort |
Rantasalo, Anssi |
title |
Novel genetic tools that enable highly pure protein production in Trichoderma reesei |
title_short |
Novel genetic tools that enable highly pure protein production in Trichoderma reesei |
title_full |
Novel genetic tools that enable highly pure protein production in Trichoderma reesei |
title_fullStr |
Novel genetic tools that enable highly pure protein production in Trichoderma reesei |
title_full_unstemmed |
Novel genetic tools that enable highly pure protein production in Trichoderma reesei |
title_sort |
novel genetic tools that enable highly pure protein production in trichoderma reesei |
publishDate |
2019 |
url |
https://cris.vtt.fi/en/publications/90d0ad99-2abc-4248-916b-bea97ec91d88 https://doi.org/10.1038/s41598-019-41573-8 http://www.scopus.com/inward/record.url?scp=85063346796&partnerID=8YFLogxK |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Rantasalo , A , Vitikainen , M , Paasikallio , T , Jäntti , J , Landowski , C P & Mojzita , D 2019 , ' Novel genetic tools that enable highly pure protein production in Trichoderma reesei ' , Scientific Reports , vol. 9 , no. 1 , 5032 . https://doi.org/10.1038/s41598-019-41573-8 |
op_relation |
https://cris.vtt.fi/en/publications/90d0ad99-2abc-4248-916b-bea97ec91d88 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1038/s41598-019-41573-8 |
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Scientific Reports |
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1800741058426961920 |