Bacillus subtilis Engineering Service
Bacillus subtilis: A Model Industrial Host
As synthetic biology continues to advance, CD BioGlyco provides a variety of efficient GlycoChas™ Cells Construction and Chassis Development services such as Plant Chassis, Microalgae Chassis, Insect Chassis, Fungal Chassis, Bacteria Chassis, and Mammalian Chassis to facilitate Biomanufacturing. B. subtilis is an important workhorse for industrial bioproduction, and with its modification by synthetic biology's gene editing tools and metabolic engineering, we provide clients with high-quality mutant strains to meet their production needs. Moreover, combining various engineering techniques such as systems biology, synthetic biology, and evolutionary engineering to construct superior biosynthetic pathways for our clients in B. subtilis.
Excellent Bacillus subtilis Engineering Service Rely on Professional Team
B. subtilis has a good expression and secretion system, and we provide the following professional engineering services:
- Strains and growth conditions
Specific genes in B. subtilis interfere and hinder its growth efficiency, we use homologous recombination to knock out certain genes individually to promote the growth of B. subtilis. Moreover, we provide productivity measurement services by detecting intracellular products, extracellular products, and recombinant proteins. With the help of adaptive laboratory evolution (ALE), we provide growth rate optimization services to obtain evolved strains.
- Precise regulation
Plasmid construction
Our lab has established plasmid construction and evaluation services to meet client needs. We mainly validate the constructed plasmids by DNA sequencing.
Genetic manipulation
CD BioGlyco provides three commonly used scarless genetic manipulation systems including site-specific recombination systems (SSR), counterselective markers (CSM), and clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated nucleases 9 (CRISPR/Cas9) systems. With SSR, we offer precise excision of DNA fragments. CRISPR-Cas9 has up to 100% two-frame in-frame knockout efficiency, the ability to introduce multipoint mutations (up to 6), and 100% efficiency, and activate and/or repress multiple genes simultaneously. CD BioGlyco controls at multiple levels to enable precise control of key points in metabolic pathways.
- Gene expression
To improve the gene expression level in B. subtilis, we serve to regulate and modify promoters at the transcriptional level. Expression of heterologous genes in B. subtilis often uses inducible and constitutive promoters.
Our researchers provide the core region of promoter modification services to enhance transcriptional activity.
Plasmids with different copy numbers also affect gene expression to some extent.
We modify promoters of different strengths and adjust gene expression to levels appropriate for specific metabolic pathways.
We provide transcriptome analysis of the genes involved in the metabolic pathways.
Publication
Technology: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE,) DNA sequencing, Sequence and ligation-independent cloning (SLIC), Enzyme-linked immunosorbent assay (ELISA), Polymerase chain reaction (PCR)
Journal: Microbial Cell Factories
Published: 2016
IF: 6.4
Results: To achieve large-scale production of B. subtilis strains, the researchers optimized the PsrfA-mediated expression system and developed the mutant strain BSG1682, which regulates gene expression in metabolic engineering by enhancing the PsrfA promoter and regulating the target genes. Moreover, the researchers established a library of derivatives exclusively for PsrfA. With the optimized BSG1682, aminopeptidase (AP) and natriuretic kinase (NK) were successfully expressed and produced to confirm the utility of the new expression system.
Fig.1 Analytical mapping of two dual promoters and their derivatives. (Guan, et al., 2016)
Applications
- Universal cell factory: B. subtilis engineering strain is a universal cell factory for industry, agriculture, biomaterials, and medicine.
- Biosensor: B. subtilis engineering strain can be used as a high-sensitivity biosensor to transform cells with different levels of control.
- Promising chassis: B. subtilis engineering strain has the ability to biosynthesize a wide range of value-added products such as pyridoxine, endoglucanase, vitamins, antibiotics, alkaline proteases, activin single-chain antibodies, nucleotides, biosurfactants, carbohydrates, and biodegradable polymers.
Frequently Asked Questions
- Why choose B. subtilis as a chassis cell?
- B. subtilis has a good fermentation base and is suitable for simple and rapid cultivation in large-scale fermentation.
- B. subtilis is a Gram-positive soil bacterium with a recognized safety profile (GRAS), a superior protein secretion mechanism, and a high-density growth profile.
- B. subtilis is non-pathogenic and environmentally compatible and able to secrete enzymes and antibiotics. In addition, it forms competent cells to take up exogenous genes and has a good expression and secretion system.
- B. subtilis has a high genetic engineering adaptability and no obvious codon bias. Thus used as a standard host strain.
- How to choose the right chassis?
- For better selection and development of suitable strains as chassis cells, it is generally necessary to consider the following factors such as feasibility of large-scale culture, culture conditions, growth rate, metabolic resources (e.g., precursors and cofactors), safety, secretion ability, ease of genetic manipulation and appropriate protein folding function.
Based on directed evolution and random mutation, CD BioGlyco provides high-quality mutants that meet the production demands. We are committed to becoming the first choice for our clients in chassis construction and development. Please feel free to contact us.
Reference
- Guan, C.; et al. Development of an efficient autoinducible expression system by promoter engineering in Bacillus subtilis. Microbial Cell Factories. 2016, 15(1): 1-12.
For research use only. Not intended for any clinical use.
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