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Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709
BACKGROUND: Bacillus licheniformis 2709 is extensively applied as a host for the high-level production of heterologous proteins, but Bacillus cells often possess unfavorable wild-type properties, such as production of viscous materials and foam during fermentation, which seriously influenced the app...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7041084/ https://www.ncbi.nlm.nih.gov/pubmed/32093734 http://dx.doi.org/10.1186/s12934-020-01307-2 |
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author | Zhou, Cuixia Zhou, Huiying Li, Dengke Zhang, Huitu Wang, Hongbin Lu, Fuping |
author_facet | Zhou, Cuixia Zhou, Huiying Li, Dengke Zhang, Huitu Wang, Hongbin Lu, Fuping |
author_sort | Zhou, Cuixia |
collection | PubMed |
description | BACKGROUND: Bacillus licheniformis 2709 is extensively applied as a host for the high-level production of heterologous proteins, but Bacillus cells often possess unfavorable wild-type properties, such as production of viscous materials and foam during fermentation, which seriously influenced the application in industrial fermentation. How to develop it from a soil bacterium to a super-secreting cell factory harboring less undomesticated properties always plays vital role in industrial production. Besides, the optimal expression pattern of the inducible enzymes like alkaline protease has not been optimized by comparing the transcriptional efficiency of different plasmids and genomic integration sites in B. licheniformis. RESULT: Bacillus licheniformis 2709 was genetically modified by disrupting the native lchAC genes related to foaming and the eps cluster encoding the extracellular mucopolysaccharide via a markerless genome-editing method. We further optimized the expression of the alkaline protease gene (aprE) by screening the most efficient expression system among different modular plasmids and genomic loci. The results indicated that genomic expression of aprE was superior to plasmid expression and finally the transcriptional level of aprE greatly increased 1.67-fold through host optimization and chromosomal integration in the vicinity of the origin of replication, while the enzyme activity significantly improved 62.19% compared with the wild-type alkaline protease-producing strain B. licheniformis. CONCLUSION: We successfully engineered an AprE high-yielding strain free of undesirable properties and its fermentation traits could be applied to bulk-production by host genetic modification and expression optimization. In summary, host optimization is an enabling technology for improving enzyme production by eliminating the harmful traits of the host and optimizing expression patterns. We believe that these strategies can be applied to improve heterologous protein expression in other Bacillus species. [Image: see text] |
format | Online Article Text |
id | pubmed-7041084 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-70410842020-03-02 Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709 Zhou, Cuixia Zhou, Huiying Li, Dengke Zhang, Huitu Wang, Hongbin Lu, Fuping Microb Cell Fact Research BACKGROUND: Bacillus licheniformis 2709 is extensively applied as a host for the high-level production of heterologous proteins, but Bacillus cells often possess unfavorable wild-type properties, such as production of viscous materials and foam during fermentation, which seriously influenced the application in industrial fermentation. How to develop it from a soil bacterium to a super-secreting cell factory harboring less undomesticated properties always plays vital role in industrial production. Besides, the optimal expression pattern of the inducible enzymes like alkaline protease has not been optimized by comparing the transcriptional efficiency of different plasmids and genomic integration sites in B. licheniformis. RESULT: Bacillus licheniformis 2709 was genetically modified by disrupting the native lchAC genes related to foaming and the eps cluster encoding the extracellular mucopolysaccharide via a markerless genome-editing method. We further optimized the expression of the alkaline protease gene (aprE) by screening the most efficient expression system among different modular plasmids and genomic loci. The results indicated that genomic expression of aprE was superior to plasmid expression and finally the transcriptional level of aprE greatly increased 1.67-fold through host optimization and chromosomal integration in the vicinity of the origin of replication, while the enzyme activity significantly improved 62.19% compared with the wild-type alkaline protease-producing strain B. licheniformis. CONCLUSION: We successfully engineered an AprE high-yielding strain free of undesirable properties and its fermentation traits could be applied to bulk-production by host genetic modification and expression optimization. In summary, host optimization is an enabling technology for improving enzyme production by eliminating the harmful traits of the host and optimizing expression patterns. We believe that these strategies can be applied to improve heterologous protein expression in other Bacillus species. [Image: see text] BioMed Central 2020-02-24 /pmc/articles/PMC7041084/ /pubmed/32093734 http://dx.doi.org/10.1186/s12934-020-01307-2 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Zhou, Cuixia Zhou, Huiying Li, Dengke Zhang, Huitu Wang, Hongbin Lu, Fuping Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709 |
title | Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709 |
title_full | Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709 |
title_fullStr | Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709 |
title_full_unstemmed | Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709 |
title_short | Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709 |
title_sort | optimized expression and enhanced production of alkaline protease by genetically modified bacillus licheniformis 2709 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7041084/ https://www.ncbi.nlm.nih.gov/pubmed/32093734 http://dx.doi.org/10.1186/s12934-020-01307-2 |
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