Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007
Abstract The production of industrial enzymes requires an efficient expression system with a suitable host. This study investigated the isolated Bacillus subtilis 007 as a host for expressing three enzymes with potential application in the food industry. Firstly, testing the PaprE and P43 promoters...
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SpringerOpen
2025-02-01
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| Series: | AMB Express |
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| Online Access: | https://doi.org/10.1186/s13568-025-01832-6 |
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| author | Jana Senger Adriana Schulz Ines Seitl Martin Heider Lutz Fischer |
| author_facet | Jana Senger Adriana Schulz Ines Seitl Martin Heider Lutz Fischer |
| author_sort | Jana Senger |
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| description | Abstract The production of industrial enzymes requires an efficient expression system with a suitable host. This study investigated the isolated Bacillus subtilis 007 as a host for expressing three enzymes with potential application in the food industry. Firstly, testing the PaprE and P43 promoters and the corresponding 5’ untranslated regions revealed great differences in the production of the recently discovered β-galactosidase from Paenibacillus wnnyii. Expression controlled by the PaprE promoter yielded a significantly higher activity of 2515 µkat/L, compared to 56 µkat/L with the P43 promoter. Modifications on the PaprE core promoter region or the spacer, the sequence between the Shine-Dalgarno sequence and the start codon, did not improve β-galactosidase production. Since the aprE 5’ untranslated region contributes to a high mRNA stability, it was incorporated into the P43 construct to determine whether mRNA stability is responsible for the differences observed in β-galactosidase production. Interestingly, mRNA stability was significantly improved and led to a nearly 50-fold higher β-galactosidase production of 2756 µkat/L. This strategy was successfully validated by the expression of two other enzymes: the cellobiose-2-epimerase from Caldicellulosiruptor saccharolyticus and the β-glucosidase from Pyrococcus furiosus. These findings underscored the crucial role of post-transcriptional regulation and emphasized mRNA stability as a key role in recombinant enzyme production in B. subtilis 007. |
| format | Article |
| id | doaj-art-ec5f9d8ca49e4e079949b043f6b6bcf4 |
| institution | Kabale University |
| issn | 2191-0855 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | SpringerOpen |
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| spelling | doaj-art-ec5f9d8ca49e4e079949b043f6b6bcf42025-02-09T12:55:51ZengSpringerOpenAMB Express2191-08552025-02-0115111210.1186/s13568-025-01832-6Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007Jana Senger0Adriana Schulz1Ines Seitl2Martin Heider3Lutz Fischer4Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of HohenheimInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of HohenheimInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of HohenheimInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of HohenheimInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of HohenheimAbstract The production of industrial enzymes requires an efficient expression system with a suitable host. This study investigated the isolated Bacillus subtilis 007 as a host for expressing three enzymes with potential application in the food industry. Firstly, testing the PaprE and P43 promoters and the corresponding 5’ untranslated regions revealed great differences in the production of the recently discovered β-galactosidase from Paenibacillus wnnyii. Expression controlled by the PaprE promoter yielded a significantly higher activity of 2515 µkat/L, compared to 56 µkat/L with the P43 promoter. Modifications on the PaprE core promoter region or the spacer, the sequence between the Shine-Dalgarno sequence and the start codon, did not improve β-galactosidase production. Since the aprE 5’ untranslated region contributes to a high mRNA stability, it was incorporated into the P43 construct to determine whether mRNA stability is responsible for the differences observed in β-galactosidase production. Interestingly, mRNA stability was significantly improved and led to a nearly 50-fold higher β-galactosidase production of 2756 µkat/L. This strategy was successfully validated by the expression of two other enzymes: the cellobiose-2-epimerase from Caldicellulosiruptor saccharolyticus and the β-glucosidase from Pyrococcus furiosus. These findings underscored the crucial role of post-transcriptional regulation and emphasized mRNA stability as a key role in recombinant enzyme production in B. subtilis 007.https://doi.org/10.1186/s13568-025-01832-6Bacillus subtilisExpressionEnzymes5’untranslated region |
| spellingShingle | Jana Senger Adriana Schulz Ines Seitl Martin Heider Lutz Fischer Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007 AMB Express Bacillus subtilis Expression Enzymes 5’untranslated region |
| title | Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007 |
| title_full | Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007 |
| title_fullStr | Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007 |
| title_full_unstemmed | Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007 |
| title_short | Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007 |
| title_sort | importance of the 5 untranslated region for recombinant enzyme production in isolated bacillus subtilis 007 |
| topic | Bacillus subtilis Expression Enzymes 5’untranslated region |
| url | https://doi.org/10.1186/s13568-025-01832-6 |
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