ArticleMicrobial cell factories2011
Regulation of mtl operon promoter of Bacillus subtilis: requirements of its use in expression vectors.
Article in Microbial cell factories, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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18 citing papers in PubMed.
- CRP-dependent DNA anchoring and membrane sequestration define a dual mechanism for MtlR-mediated regulation.Nucleic acids research · 2026Article
- Engineering Bacillus subtilis for high-value bioproduction: recent advances and applications.Microbial cell factories · 2025Review
- AdvancingMicroorganisms · 2024Review
- Biotechnological and food synthetic biology potential of platform strain:Synthetic and systems biotechnology · 2023Review
- An operator-based expression toolkit forProceedings of the National Academy of Sciences of the United States of America · 2022Article
- The multifunctionality of expression systems inExperimental biology and medicine (Maywood, N.J.) · 2021Review
- Deciphering the Regulation of the Mannitol Operon Paves the Way for Efficient Production of Mannitol in Lactococcus lactis.Applied and environmental microbiology · 2021Article
- Article
- Maltose Induced Expression of Cecropin AD by SUMO Technology in Bacillus subtilis WB800N.The protein journal · 2020Article
- Phosphosugar Stress in Bacillus subtilis: Intracellular Accumulation of Mannose 6-Phosphate Derepressed theJournal of bacteriology · 2019Article
- Exploitation of Bacillus subtilis as a robust workhorse for production of heterologous proteins and beyond.World journal of microbiology & biotechnology · 2018Review
- Genome-Based Genetic Tool Development forFrontiers in microbiology · 2016Article
- An Alternative Bacterial Expression System UsingIranian journal of biotechnology · 2015Article
- Complete Genome Sequence of Bacillus subtilis subsp. subtilis Strain 3NA.Genome announcements · 2015Article
- The bacterial phosphoenolpyruvate:carbohydrate phosphotransferase system: regulation by protein phosphorylation and phosphorylation-dependent protein-protein interactions.Microbiology and molecular biology reviews : MMBR · 2014Review
- Current development in genetic engineering strategies of Bacillus species.Microbial cell factories · 2014Review
- Subtilase SprP exerts pleiotropic effects in Pseudomonas aeruginosa.MicrobiologyOpen · 2014Article
- Generation of an artificial double promoter for protein expression in Bacillus subtilis through a promoter trap system.PloS one · 2013Article
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3 authors.
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Abstract
backgroundSeveral vector systems have been developed to express any gene desired to be studied in Bacillus subtilis. Among them, the transcriptionally regulated promoters involved in carbohydrate utilization are a research priority. Expression systems based on Bacillus promoters for xylose, maltose, and mannose utilization, as well as on the heterologous E. coli lactose promoter, have been successfully constructed. The promoter of the mtlAFD operon for utilization of mannitol is another promising candidate for its use in expression vectors. In this study, we investigated the regulation of the mtl genes in order to identify the elements needed to construct a strong mannitol inducible expression system in B. subtilis.
resultsRegulation of the promoters of mtlAFD operon (P(mtlA)) and mtlR (P(mtlR)) encoding the activator were investigated by fusion to lacZ. Identification of the P(mtlA) and P(mtlR) transcription start sites revealed the σ(A) like promoter structures. Also, the operator of P(mtlA) was determined by shortening, nucleotide exchange, and alignment of P(mtlA) and P(mtlR) operator regions. Deletion of the mannitol-specific PTS genes (mtlAF) resulted in P(mtlA) constitutive expression demonstrating the inhibitory effect of EIICB(Mtl) and EIIA(Mtl) on MtlR in the absence of mannitol. Disruption of mtlD made the cells sensitive to mannitol and glucitol. Both P(mtlA) and P(mtlR) were influenced by carbon catabolite repression (CCR). However, a CcpA deficient mutant showed only a slight reduction in P(mtlR) catabolite repression. Similarly, using P(groE) as a constitutive promoter, putative cre sites of P(mtlA) and P(mtlR) slightly reduced the promoter activity in the presence of glucose. In contrast, glucose repression of P(mtlA) and P(mtlR) was completely abolished in a ΔptsG mutant and significantly reduced in a MtlR (H342D) mutant.
conclusionsThe mtl operon promoter (P(mtlA)) is a strong promoter that reached a maximum of 13,000 Miller units with lacZ as a reporter on low copy plasmids. It is tightly regulated by just one copy of the mtlR gene on the chromosome and subject to CCR. CCR can be switched off by mutations in MtlR and the glucose transporter. These properties and the low costs of the inducers, i.e. mannitol and glucitol, make the promoter ideal for designing regulated expression systems.
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