Transcriptome based functional identification and application of regulator AbrB on alkaline protease synthesis in Bacillus licheniformis 2709
| Autor: | Huiying Zhou, Fuping Lu, Honglei Fang, Sun Yanqing, Cuixia Zhou, Huitu Zhang, Guangcheng Yang |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Mutant
Repressor 02 engineering and technology Biochemistry Industrial Microbiology 03 medical and health sciences Bacterial Proteins Structural Biology Transcription (biology) Bacillus licheniformis Binding site Molecular Biology Gene 030304 developmental biology chemistry.chemical_classification Regulation of gene expression 0303 health sciences biology Chemistry Membrane Transport Proteins Gene Expression Regulation Bacterial General Medicine 021001 nanoscience & nanotechnology biology.organism_classification Enzyme Transcriptome 0210 nano-technology Transcription Factors |
| Zdroj: | International Journal of Biological Macromolecules. 166:1491-1498 |
| ISSN: | 0141-8130 |
| DOI: | 10.1016/j.ijbiomac.2020.11.028 |
| Popis: | Bacillus licheniformis 2709 is the major alkaline protease producer, which has great potential value of industrial application, but how the high-producer can be regulated rationally is still not completely understood. It's meaningful to understand the metabolic processes during alkaline protease production in industrial fermentation medium. Here, we collected the transcription database at various enzyme-producing stages (preliminary stage, stable phase and decline phase) to specifically research the synthesized and regulatory mechanism of alkaline protease in B. licheniformis. The RNA-sequencing analysis showed differential expression of numerous genes related to several processes, among which genes correlated with regulators were concerned, especially the major differential gene abrB on enzyme (AprE) synthesis was investigated. It was further verified that AbrB is a repressor of AprE by plasmid-mediated over-expression due to the severely descending enzyme activity (11,300 U/mL to 2695 U/mL), but interestingly it is indispensable for alkaline protease production because the enzyme activity of the null abrB mutant was just about 2279 U/mL. Thus, we investigated the aprE transcription by eliminating the theoretical binding site (TGGAA) of AbrB protein predicated by computational strategy, which significantly improved the enzyme activity by 1.21-fold and gene transcription level by 1.77-fold in the mid-log phase at a cultivation time of 18 h. Taken together, it is of great significance to improve the production strategy, control the metabolic process and oriented engineering by rational molecular modification of regulatory network based on the high throughput sequencing and computational prediction. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect