Actinomycetes Engineering Service
High-yield, High-quality, and High-stability Actinomycetes Engineering strategy
With synthetic biology and metabolic engineering, CD BioGlyco provides professional GlycoChas™ Cells and Chassis Development. Our lab provides new natural product Production and engineering strain development of Actinomycetes. Actinomycetes is a Gram-positive Bacterium and has abundant genomic and morphological/physiological features. The biosynthetic mechanism of actinobacteria secondary metabolites includes elements such as synthetic enzymes, tailoring enzymes, regulatory enzymes, and transporters, and the respective coding genes usually cluster together in the genome to form biosynthetic gene clusters (BGCs). We provide a range of services including strain improvement, precursor engineering, and gene regulation in Actinomycetes.
- Strain improvement
Actinomycetes are an excellent host for genetic manipulation, and we offer a variety of services to enhance heterologous expression and modify biosynthetic pathways through promoter reconstruction, global pathway regulation, and gene modification. We introduce gene modifications through recombinant DNA technology to redirect to ensure high yields. Moreover, we improve high-titer strain engineering via mutation and screening. For the determination of mycelial growth, we estimate mycelial growth by testing dry weights.
Multiplexed site-specific genome engineering
Combining the in vitro clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated nucleases 9 (CRISPR/Cas9) system with Gibson assembly, CD BioGlyco provides professional BGC refactoring and mutant construction services. We use a novel in vitro DNA editing method to construct multiple recombinant plasmids containing different plasmids with key factor deletions and strong promoter substitutions. Then, they are ligated by polymerase chain reaction (PCR) and overlapping PCR, respectively, using primers. Mutant strains are constructed in our lab using CRISPR/Cas9 genome editing tools.
Fig.1 Schematic diagram of Actinomycetes engineering service. (CD BioGlyco)
Publication
Technology: Transcriptome sequencing, Gene deletion and overexpression, Phase-contrast microscopy, Laser confocal microscopy, High-performance liquid chromatography (HPLC), Electrophoretic mobility shift assay
Journal: Biomolecules
Published: 2020
IF: 6.064
Results: In this work, researchers analyzed the biological yield and mycelial fragmentation by constructing APASM_4178 gene deletion and overexpression strains. APASM_4178 is a coding gene which transcription is controlled by a specific adpa-like protein. The gene deletion strain showed 43.65% higher yield compared to the original strain. Thus, APASM_4178 is an effective factor for morphological engineering and antibiotic yield in actinomycetes.
Fig.2 Expression and analysis profiles of gene deletion strains. (Wu, et al., 2020)
Applications
- Actinomycetes engineering can be used to produce a variety of high-titer drug lead compounds.
- Actinomycetes engineering for the discovery and development of novel bioactive secondary metabolites of Actinomycetes.
Frequently Asked Questions
- How to overproduce specific Actinomycete secondary metabolites?
- Regulation of specific biosynthetic pathways
- Alter the metabolic flux distribution of their different precursors
- Structural genes coding for overexpression of enzymes involved in metabolite biosynthesis
- Increase self-resistance or induce resistance to several antibiotics
- Expression of biosynthetic gene clusters in heterologous hosts or industrially optimized strains
- Metabolic engineering is an effective strategy for optimizing secondary metabolism in actinomycetes, and what metabolic engineering commonly used for production includes?
- Overexpression or disruption of pleiotropic / pathway-specific regulatory genes
- Enhanced precursor supply
- Modify the transcription/translation apparatus
- Replication or higher-order amplification of rate-limiting enzyme-encoding genes or intact BGCs
CD BioGlyco has a professional technical team and superior chassis cells to help clients improve the product yield and improve the genetic stability of engineered strains. Our staff has quite some experience in chassis construction and development. Please feel free to contact us.
Reference
- Wu, Y.; et al. Subtilisin-involved morphology engineering for improved antibiotic production in actinomycetes. Biomolecules. 2020, 10(6): 851.
For research use only. Not intended for any clinical use.
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