Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass
Abstract Background Fermentation of sugars derived from plant biomass feedstock is crucial for sustainability. Hence, utilizing customized enzymatic cocktails to obtain oligosaccharides instead of monomers is an alternative fermentation strategy to produce prebiotics, cosmetics, and biofuels. This s...
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BMC
2024-11-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-024-02578-9 |
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| author | Fernanda Lopes de Figueiredo Fabiano Jares Contesini César Rafael Fanchini Terrasan Jaqueline Aline Gerhardt Ana Beatriz Corrêa Everton Paschoal Antoniel Natália Sayuri Wassano Lucas Levassor Sarita Cândida Rabelo Telma Teixeira Franco Uffe Hasbro Mortensen André Damasio |
| author_facet | Fernanda Lopes de Figueiredo Fabiano Jares Contesini César Rafael Fanchini Terrasan Jaqueline Aline Gerhardt Ana Beatriz Corrêa Everton Paschoal Antoniel Natália Sayuri Wassano Lucas Levassor Sarita Cândida Rabelo Telma Teixeira Franco Uffe Hasbro Mortensen André Damasio |
| author_sort | Fernanda Lopes de Figueiredo |
| collection | DOAJ |
| description | Abstract Background Fermentation of sugars derived from plant biomass feedstock is crucial for sustainability. Hence, utilizing customized enzymatic cocktails to obtain oligosaccharides instead of monomers is an alternative fermentation strategy to produce prebiotics, cosmetics, and biofuels. This study developed an engineered strain of Aspergillus niger producing a tailored cellulolytic cocktail capable of partially degrading sugarcane straw to yield cellooligosaccharides. Results The A. niger prtT∆ strain created resulted in a reduced extracellular protease production. The prtT∆ background was then used to create strains by deleting exoenzyme encoding genes involved in mono- or disaccharide formation. Consequently, we successfully generated a tailored prtT∆bglA∆ strain by eliminating a beta-glucosidase (bglA) gene and subsequently deleted two cellobiohydrolases and one beta-xylosidase encoding genes using a multiplex strategy, resulting in the Quintuple∆ strain (prtT∆; bglA∆; cbhA∆; cbhB∆; xlnD∆). When applied for sugarcane biomass degradation, the tailored secretomes produced by A. niger resulted in a higher ratio of cellobiose and cellotriose compared with glucose relative to the reference strain. Mass spectrometry revealed that the Quintuple∆ strain secreted alternative cellobiohydrolases and beta-glucosidases to compensate for the absence of major cellulases. Enzymes targeting minor polysaccharides in plant biomass were also upregulated in this tailored strain. Conclusion Tailored secretome use increased COS/glucose ratio during sugarcane biomass degradation showing that deleting some enzymatic components is an effective approach for producing customized enzymatic cocktails. Our findings highlight the plasticity of fungal genomes as enzymes that target minor components of plant cell walls, and alternative cellulases were produced by the mutant strain. Despite deletion of important secretome components, fungal growth was maintained in plant biomass. |
| format | Article |
| id | doaj-art-a9d4176d3b774fdb86664d6b0ea92e54 |
| institution | OA Journals |
| issn | 1475-2859 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
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| series | Microbial Cell Factories |
| spelling | doaj-art-a9d4176d3b774fdb86664d6b0ea92e542025-08-20T02:38:42ZengBMCMicrobial Cell Factories1475-28592024-11-0123111310.1186/s12934-024-02578-9Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomassFernanda Lopes de Figueiredo0Fabiano Jares Contesini1César Rafael Fanchini Terrasan2Jaqueline Aline Gerhardt3Ana Beatriz Corrêa4Everton Paschoal Antoniel5Natália Sayuri Wassano6Lucas Levassor7Sarita Cândida Rabelo8Telma Teixeira Franco9Uffe Hasbro Mortensen10André Damasio11Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP)Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP)Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP)Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP)Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP)Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP)Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP)Department of Biotechnology and Biomedicine, Technical University of Denmark (DTU)Department of Bioprocess and Biotechnology, School of Agricultural Sciences, São Paulo State University (UNESP)Interdisciplinary Center of Energy Planning, Universidade Estadual de Campinas (UNICAMP)Department of Biotechnology and Biomedicine, Technical University of Denmark (DTU)Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP)Abstract Background Fermentation of sugars derived from plant biomass feedstock is crucial for sustainability. Hence, utilizing customized enzymatic cocktails to obtain oligosaccharides instead of monomers is an alternative fermentation strategy to produce prebiotics, cosmetics, and biofuels. This study developed an engineered strain of Aspergillus niger producing a tailored cellulolytic cocktail capable of partially degrading sugarcane straw to yield cellooligosaccharides. Results The A. niger prtT∆ strain created resulted in a reduced extracellular protease production. The prtT∆ background was then used to create strains by deleting exoenzyme encoding genes involved in mono- or disaccharide formation. Consequently, we successfully generated a tailored prtT∆bglA∆ strain by eliminating a beta-glucosidase (bglA) gene and subsequently deleted two cellobiohydrolases and one beta-xylosidase encoding genes using a multiplex strategy, resulting in the Quintuple∆ strain (prtT∆; bglA∆; cbhA∆; cbhB∆; xlnD∆). When applied for sugarcane biomass degradation, the tailored secretomes produced by A. niger resulted in a higher ratio of cellobiose and cellotriose compared with glucose relative to the reference strain. Mass spectrometry revealed that the Quintuple∆ strain secreted alternative cellobiohydrolases and beta-glucosidases to compensate for the absence of major cellulases. Enzymes targeting minor polysaccharides in plant biomass were also upregulated in this tailored strain. Conclusion Tailored secretome use increased COS/glucose ratio during sugarcane biomass degradation showing that deleting some enzymatic components is an effective approach for producing customized enzymatic cocktails. Our findings highlight the plasticity of fungal genomes as enzymes that target minor components of plant cell walls, and alternative cellulases were produced by the mutant strain. Despite deletion of important secretome components, fungal growth was maintained in plant biomass.https://doi.org/10.1186/s12934-024-02578-9Aspergillus nigerCAZymesCellooligosaccharidesFungal engineeringTailor-made enzymatic cocktailSugarcane biomass |
| spellingShingle | Fernanda Lopes de Figueiredo Fabiano Jares Contesini César Rafael Fanchini Terrasan Jaqueline Aline Gerhardt Ana Beatriz Corrêa Everton Paschoal Antoniel Natália Sayuri Wassano Lucas Levassor Sarita Cândida Rabelo Telma Teixeira Franco Uffe Hasbro Mortensen André Damasio Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass Microbial Cell Factories Aspergillus niger CAZymes Cellooligosaccharides Fungal engineering Tailor-made enzymatic cocktail Sugarcane biomass |
| title | Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass |
| title_full | Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass |
| title_fullStr | Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass |
| title_full_unstemmed | Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass |
| title_short | Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass |
| title_sort | engineering the secretome of aspergillus niger for cellooligosaccharides production from plant biomass |
| topic | Aspergillus niger CAZymes Cellooligosaccharides Fungal engineering Tailor-made enzymatic cocktail Sugarcane biomass |
| url | https://doi.org/10.1186/s12934-024-02578-9 |
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