Broadly applicable genetic tools for fungi
Engineering of microbial hosts for industrial use has became significantly easier and faster in the recent years. This has been enabled by novel molecular biology tools and genome editing methods. However, the gene expression tools required for optimization of the engineered organisms are often inef...
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Format: | Doctoral or Postdoctoral Thesis |
Language: | English |
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Aalto University
2019
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Online Access: | https://aaltodoc.aalto.fi/handle/123456789/36182 |
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Aalto University Publication Archive (Aaltodoc) |
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English |
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Chemistry expression system yeast fungi synthetic biology transcription factor industrial biotechnology ekspressiosysteemi hiiva home synteettinen biologia transkriptiofaktori teollinen biotekniikka |
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Chemistry expression system yeast fungi synthetic biology transcription factor industrial biotechnology ekspressiosysteemi hiiva home synteettinen biologia transkriptiofaktori teollinen biotekniikka Rantasalo, Anssi Broadly applicable genetic tools for fungi |
topic_facet |
Chemistry expression system yeast fungi synthetic biology transcription factor industrial biotechnology ekspressiosysteemi hiiva home synteettinen biologia transkriptiofaktori teollinen biotekniikka |
description |
Engineering of microbial hosts for industrial use has became significantly easier and faster in the recent years. This has been enabled by novel molecular biology tools and genome editing methods. However, the gene expression tools required for optimization of the engineered organisms are often inefficient, or completely lacking especially in less-established production hosts. This hinders the rapid development of novel biotechnological applications. This PhD work focused on developing synthetic biology tools that allow accurate control of gene expression in a broad spectrum of fungal species. The outcomes of this thesis represent an important advance in the field of biological engineering because fungi comprise an industrially important group of organisms. The developed tools provide high utility especially in applications in which precise balancing or strong gene expression is required. In the first part of this work (Publication I and II), a library of modular genetic parts, including a set of core promoters and synthetic transcription factors was established in Saccharomyces cerevisiae. These tools enable construction of complex genetic devices and they provided a broad gene expression range. Next part of this work focuses on development of universal gene expression system for fungi (Publication III). The functionality of the system was demonstrated in eight diverse fungal hosts: in six yeasts and two filamentous fungi. Importantly, due to universal performance of the system, these tools also makes it possible to introduce novel eukaryotic microbes for biotechnological use. In the final part, the developed synthetic tools were applied in a protein production application (Publication IV). The selected example protein product was lipase B of Candida antarctica (calB), and the production was carried out in filamentous fungus Trichoderma reesei. The obtained protein levels were comparable to the levels obtained by using the commonly employed inducible cbh1 promoter. In addition, due to highly constitutive ... |
author2 |
Mojzita, Dominik, Dr., VTT Technical Research Centre of Finland, Finland Jäntti, Jussi, Dr., VTT Technical Research Centre of Finland, Finland Kemian tekniikan korkeakoulu School of Chemical Technology Biotuotteiden ja biotekniikan laitos Department of Bioproducts and Biosystems Frey, Alexander, Assoc. Prof., Aalto University, Department of Bioproducts and Biosystems, Finland Aalto-yliopisto Aalto University |
format |
Doctoral or Postdoctoral Thesis |
author |
Rantasalo, Anssi |
author_facet |
Rantasalo, Anssi |
author_sort |
Rantasalo, Anssi |
title |
Broadly applicable genetic tools for fungi |
title_short |
Broadly applicable genetic tools for fungi |
title_full |
Broadly applicable genetic tools for fungi |
title_fullStr |
Broadly applicable genetic tools for fungi |
title_full_unstemmed |
Broadly applicable genetic tools for fungi |
title_sort |
broadly applicable genetic tools for fungi |
publisher |
Aalto University |
publishDate |
2019 |
url |
https://aaltodoc.aalto.fi/handle/123456789/36182 |
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Antarc* Antarctica |
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Antarc* Antarctica |
op_relation |
Aalto University publication series DOCTORAL DISSERTATIONS 2/2019 [Publication 1]: Rantasalo A., Czeizler E., Virtanen R., Rousu J., Lähdesmäki H., Penttilä M., Jäntti J., Mojzita D. Synthetic transcription amplifier system for orthogonal control of gene expression in Saccharomyces cerevisiae. PLoS One, 2016, 11, e0148320. Full Text in Acris/Aaltodoc: http://urn.fi/URN:NBN:fi:aalto-201705114007. DOI:10.1371/journal.pone.0148320 [Publication 2]: Rantasalo A., Kuivanen J., Penttilä M., Jäntti J., Mojzita D. Synthetic Toolkit for Complex Genetic Circuit Engineering in Saccharomyces cerevisiae. ACS Synthetic biology, 2018, Volume 7, 1573–1587. DOI:10.1021/acssynbio.8b00076 [Publication 3]: Rantasalo A., Landowski C.P., Kuivanen J., Korppoo A., Reuter L., Koivistoinen O., Valkonen M., Penttilä M., Jäntti J., Mojzita D. A universal gene expression system for fungi. Nucleic Acids Research, 2018, Volume 46, e111. DOI:10.1093/nar/gky558 [Publication 4]: Rantasalo A., Vitikainen M., Paasikallio T., Jäntti J., Landowski C.P., Mojzita D. Novel genetic tools that enable highly pure protein production in Trichoderma reesei. 2018, Under review in Scientific Reports. 978-952-60-8362-9 (electronic) 978-952-60-8361-2 (printed) 1799-4942 (electronic) 1799-4934 (printed) 1799-4934 (ISSN-L) https://aaltodoc.aalto.fi/handle/123456789/36182 URN:ISBN:978-952-60-8362-9 |
op_doi |
https://doi.org/10.1371/journal.pone.0148320 https://doi.org/10.1021/acssynbio.8b00076 https://doi.org/10.1093/nar/gky558 |
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PLOS ONE |
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e0148320 |
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ftaaltouniv:oai:aaltodoc.aalto.fi:123456789/36182 2023-05-15T13:34:36+02:00 Broadly applicable genetic tools for fungi Laajatoimiset geneettiset työkalut hiiva- ja homeorganismeille Rantasalo, Anssi Mojzita, Dominik, Dr., VTT Technical Research Centre of Finland, Finland Jäntti, Jussi, Dr., VTT Technical Research Centre of Finland, Finland Kemian tekniikan korkeakoulu School of Chemical Technology Biotuotteiden ja biotekniikan laitos Department of Bioproducts and Biosystems Frey, Alexander, Assoc. Prof., Aalto University, Department of Bioproducts and Biosystems, Finland Aalto-yliopisto Aalto University 2019 application/pdf https://aaltodoc.aalto.fi/handle/123456789/36182 en eng Aalto University Aalto-yliopisto Aalto University publication series DOCTORAL DISSERTATIONS 2/2019 [Publication 1]: Rantasalo A., Czeizler E., Virtanen R., Rousu J., Lähdesmäki H., Penttilä M., Jäntti J., Mojzita D. Synthetic transcription amplifier system for orthogonal control of gene expression in Saccharomyces cerevisiae. PLoS One, 2016, 11, e0148320. Full Text in Acris/Aaltodoc: http://urn.fi/URN:NBN:fi:aalto-201705114007. DOI:10.1371/journal.pone.0148320 [Publication 2]: Rantasalo A., Kuivanen J., Penttilä M., Jäntti J., Mojzita D. Synthetic Toolkit for Complex Genetic Circuit Engineering in Saccharomyces cerevisiae. ACS Synthetic biology, 2018, Volume 7, 1573–1587. DOI:10.1021/acssynbio.8b00076 [Publication 3]: Rantasalo A., Landowski C.P., Kuivanen J., Korppoo A., Reuter L., Koivistoinen O., Valkonen M., Penttilä M., Jäntti J., Mojzita D. A universal gene expression system for fungi. Nucleic Acids Research, 2018, Volume 46, e111. DOI:10.1093/nar/gky558 [Publication 4]: Rantasalo A., Vitikainen M., Paasikallio T., Jäntti J., Landowski C.P., Mojzita D. Novel genetic tools that enable highly pure protein production in Trichoderma reesei. 2018, Under review in Scientific Reports. 978-952-60-8362-9 (electronic) 978-952-60-8361-2 (printed) 1799-4942 (electronic) 1799-4934 (printed) 1799-4934 (ISSN-L) https://aaltodoc.aalto.fi/handle/123456789/36182 URN:ISBN:978-952-60-8362-9 Chemistry expression system yeast fungi synthetic biology transcription factor industrial biotechnology ekspressiosysteemi hiiva home synteettinen biologia transkriptiofaktori teollinen biotekniikka G5 Artikkeliväitöskirja text Doctoral dissertation (article-based) Väitöskirja (artikkeli) 2019 ftaaltouniv https://doi.org/10.1371/journal.pone.0148320 https://doi.org/10.1021/acssynbio.8b00076 https://doi.org/10.1093/nar/gky558 2022-12-15T19:18:24Z Engineering of microbial hosts for industrial use has became significantly easier and faster in the recent years. This has been enabled by novel molecular biology tools and genome editing methods. However, the gene expression tools required for optimization of the engineered organisms are often inefficient, or completely lacking especially in less-established production hosts. This hinders the rapid development of novel biotechnological applications. This PhD work focused on developing synthetic biology tools that allow accurate control of gene expression in a broad spectrum of fungal species. The outcomes of this thesis represent an important advance in the field of biological engineering because fungi comprise an industrially important group of organisms. The developed tools provide high utility especially in applications in which precise balancing or strong gene expression is required. In the first part of this work (Publication I and II), a library of modular genetic parts, including a set of core promoters and synthetic transcription factors was established in Saccharomyces cerevisiae. These tools enable construction of complex genetic devices and they provided a broad gene expression range. Next part of this work focuses on development of universal gene expression system for fungi (Publication III). The functionality of the system was demonstrated in eight diverse fungal hosts: in six yeasts and two filamentous fungi. Importantly, due to universal performance of the system, these tools also makes it possible to introduce novel eukaryotic microbes for biotechnological use. In the final part, the developed synthetic tools were applied in a protein production application (Publication IV). The selected example protein product was lipase B of Candida antarctica (calB), and the production was carried out in filamentous fungus Trichoderma reesei. The obtained protein levels were comparable to the levels obtained by using the commonly employed inducible cbh1 promoter. In addition, due to highly constitutive ... Doctoral or Postdoctoral Thesis Antarc* Antarctica Aalto University Publication Archive (Aaltodoc) PLOS ONE 11 2 e0148320 |