Development of <i>Aspergillus oryzae</i> BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications
Enzyme-mediated protein degradation is a major concern in industrial fungal strain improvement, making low-proteolytic strains preferable for enhanced protein production. Here, we improved food-grade <i>Aspergillus oryzae</i> BCC7051 by manipulating the transcriptional regulation of prot...
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2024-12-01
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| author | Sarocha Panchanawaporn Chanikul Chutrakul Sukanya Jeennor Jutamas Anantayanon Kobkul Laoteng |
| author_facet | Sarocha Panchanawaporn Chanikul Chutrakul Sukanya Jeennor Jutamas Anantayanon Kobkul Laoteng |
| author_sort | Sarocha Panchanawaporn |
| collection | DOAJ |
| description | Enzyme-mediated protein degradation is a major concern in industrial fungal strain improvement, making low-proteolytic strains preferable for enhanced protein production. Here, we improved food-grade <i>Aspergillus oryzae</i> BCC7051 by manipulating the transcriptional regulation of protease-encoding genes. Genome mining of the transcription factor <i>AoprtR</i> and computational analysis confirmed its deduced amino acid sequence sharing evolutionary conservation across <i>Aspergillus</i> and <i>Penicillium</i> spp. The AoPrtR protein, which is classified into the Zn(II)2-Cys6-type transcription factor family, manipulates both intra- and extracellular proteolytic enzymes. Our transcriptional analysis indicated that the regulation of several protease-encoding genes was AoPrtR-dependent, with AoPrtR acting as a potent activator for extracellular acid-protease-encoding genes and a likely repressor for intracellular non-acid-protease-encoding genes. An indirect regulatory mechanism independent of PrtR may enhance proteolysis. Moreover, AoPrtR disruption increased extracellular esterase production by 2.55-fold, emphasizing its role in protein secretion. Our findings highlight the complexity of AoPrtR-mediated regulation by <i>A. oryzae</i>. Manipulation of regulatory processes through AoPrtR prevents secreted protein degradation and enhances the quantity of extracellular proteins, suggesting the low-proteolytic variant as a promising platform for the production of these proteins. This modified strain has biotechnological potential for further refinement and sustainable production of bio-based products in the food, feed, and nutraceutical industries. |
| format | Article |
| id | doaj-art-4ece2bd6d9eb4a38abeedc3318bb7eaa |
| institution | Kabale University |
| issn | 2309-608X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj-art-4ece2bd6d9eb4a38abeedc3318bb7eaa2025-01-24T13:37:11ZengMDPI AGJournal of Fungi2309-608X2024-12-01111610.3390/jof11010006Development of <i>Aspergillus oryzae</i> BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial ApplicationsSarocha Panchanawaporn0Chanikul Chutrakul1Sukanya Jeennor2Jutamas Anantayanon3Kobkul Laoteng4Functional Ingredients and Food Innovation Research Group (IFIG), National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, ThailandFunctional Ingredients and Food Innovation Research Group (IFIG), National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, ThailandFunctional Ingredients and Food Innovation Research Group (IFIG), National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, ThailandFunctional Ingredients and Food Innovation Research Group (IFIG), National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, ThailandFunctional Ingredients and Food Innovation Research Group (IFIG), National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, ThailandEnzyme-mediated protein degradation is a major concern in industrial fungal strain improvement, making low-proteolytic strains preferable for enhanced protein production. Here, we improved food-grade <i>Aspergillus oryzae</i> BCC7051 by manipulating the transcriptional regulation of protease-encoding genes. Genome mining of the transcription factor <i>AoprtR</i> and computational analysis confirmed its deduced amino acid sequence sharing evolutionary conservation across <i>Aspergillus</i> and <i>Penicillium</i> spp. The AoPrtR protein, which is classified into the Zn(II)2-Cys6-type transcription factor family, manipulates both intra- and extracellular proteolytic enzymes. Our transcriptional analysis indicated that the regulation of several protease-encoding genes was AoPrtR-dependent, with AoPrtR acting as a potent activator for extracellular acid-protease-encoding genes and a likely repressor for intracellular non-acid-protease-encoding genes. An indirect regulatory mechanism independent of PrtR may enhance proteolysis. Moreover, AoPrtR disruption increased extracellular esterase production by 2.55-fold, emphasizing its role in protein secretion. Our findings highlight the complexity of AoPrtR-mediated regulation by <i>A. oryzae</i>. Manipulation of regulatory processes through AoPrtR prevents secreted protein degradation and enhances the quantity of extracellular proteins, suggesting the low-proteolytic variant as a promising platform for the production of these proteins. This modified strain has biotechnological potential for further refinement and sustainable production of bio-based products in the food, feed, and nutraceutical industries.https://www.mdpi.com/2309-608X/11/1/6<i>Aspergillus oryzae</i>PrtRtranscription regulatorproteaseesterasebiotechnological potential |
| spellingShingle | Sarocha Panchanawaporn Chanikul Chutrakul Sukanya Jeennor Jutamas Anantayanon Kobkul Laoteng Development of <i>Aspergillus oryzae</i> BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications Journal of Fungi <i>Aspergillus oryzae</i> PrtR transcription regulator protease esterase biotechnological potential |
| title | Development of <i>Aspergillus oryzae</i> BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications |
| title_full | Development of <i>Aspergillus oryzae</i> BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications |
| title_fullStr | Development of <i>Aspergillus oryzae</i> BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications |
| title_full_unstemmed | Development of <i>Aspergillus oryzae</i> BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications |
| title_short | Development of <i>Aspergillus oryzae</i> BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications |
| title_sort | development of i aspergillus oryzae i bcc7051 as a robust cell factory towards the transcriptional regulation of protease encoding genes for industrial applications |
| topic | <i>Aspergillus oryzae</i> PrtR transcription regulator protease esterase biotechnological potential |
| url | https://www.mdpi.com/2309-608X/11/1/6 |
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