Optimization of alkaline protease production by rational deletion of sporulation related genes in Bacillus licheniformis
| Autor: | Fuping Lu, Cuixia Zhou, Huiying Zhou, Huitu Zhang |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
medicine.medical_treatment Mutant lcsh:QR1-502 Bioengineering 01 natural sciences Applied Microbiology and Biotechnology lcsh:Microbiology Gene Knockout Techniques 03 medical and health sciences Bacterial Proteins 010608 biotechnology Endopeptidases medicine Bacillus licheniformis Alkaline protease Gene Sequence Deletion 030304 developmental biology Spores Bacterial chemistry.chemical_classification 0303 health sciences Protease biology Asporogenic mutants Chemistry Research Genetic Complementation Test fungi biology.organism_classification Enzyme assay Enzyme Biochemistry Sporulation Fermentation biology.protein Exoenzyme Biotechnology |
| Zdroj: | Microbial Cell Factories, Vol 18, Iss 1, Pp 1-12 (2019) Microbial Cell Factories |
| ISSN: | 1475-2859 |
| DOI: | 10.1186/s12934-019-1174-1 |
| Popis: | Background Our laboratory has constructed a Bacillus licheniformis strain that secretes alkaline protease (AprE) with excellent enzymatic properties. B. licheniformis is generally regarded as safe and has a high industrial exoenzyme secretion capacity, but the host retains some undomesticated characteristic that increase its competitiveness and survival, such as spore-formation, which increases the requirements and difficulties in industrial operations (e.g. sterilization and enzyme activity control). Furthermore, the influence of sporulation on alkaline protease production in B. licheniformis has not been elucidated in detail. Result A series of asporogenic variants of the parent strain were constructed by individually knocking out the master regulator genes (spo0A, sigF and sigE) involved in sporulation. Most of the variants formed abortively disporic cells characterized by asymmetric septa at the poles and unable to survive incubation at 75 °C for 10 min. Two of them (ΔsigF and ΔsigE) exhibited superior characteristics in protease production, especially improving the expression of the aprE gene. Under the currently used fermentation conditions, the vegetative production phase of ΔsigF can be prolonged to 72 h, and the highest protease production of ΔsigF reached 29,494 ± 1053 U/mL, which was about 19.7% higher than that of the wild-type strain. Conclusion We first constructed three key sporulation-deficient strain to investigate the effect of sporulation on alkaline protease synthesis. The sigF mutant retained important industrial properties such as facilitating the sterilization process, a prolonged stable phase of enzyme production and slower decreasing trend, which will be superior in energy conservation, simpler operations and target product controlling effect. In summary, the work provides a useful industrial host with preferable characteristics and a novel strategy to enhance the production of protease. Electronic supplementary material The online version of this article (10.1186/s12934-019-1174-1) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
| Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |