Microorganisms are tiny organisms that are widely found in nature and are too small and numerous to be seen directly by the naked eye. Microorganisms can be categorized into prokaryotic, eukaryotic, and non-cellular microorganisms according to their evolutionary level and traits. The number of microorganisms in nature is roughly estimated to be in the order of millions, making them a rich and diverse resource on the earth. Nowadays, microbial technology is an integral part of bioengineering. Microbial technology has been successfully applied in various industries such as medicine, food, and textile. With the further development of biotechnology, it has become easier to build non-natural biomanufacturing systems based on microorganisms. For example, microorganisms can be used to efficiently produce protein macromolecules, DNA/RNA strands, and nucleotides.
Fig.1 Different forms of microorganisms. (Wikipedia)
We utilize state-of-the-art synthetic biology and genetic engineering techniques to precisely manipulate microbial metabolic pathways. This involves introducing novel genes, optimizing enzyme expression, and diverting metabolic flux towards the desired nucleotide sugar synthesis pathways. Our sophisticated bioinformatics tools and predictive modeling enable rational design for enhanced productivity and specificity.
Our robust high-throughput screening platforms allow for rapid identification of high-performing microbial strains. Coupled with directed evolution strategies, we iteratively enhance the efficiency and yield of our engineered microbes, ensuring continuous improvement in production capabilities and expanding the library of accessible nucleotide sugars.
We employ advanced bioreactor design and process control strategies to create optimal growth and production environments for our microbial factories. This includes precise control over pH, temperature, aeration, and nutrient feeding, maximizing volumetric productivity and overall yield of the target nucleotide sugars.
Nucleotides are an indispensable class of substances for human beings, and the microbial synthesis method has the advantages of mild conditions, high specificity, high production efficiency, simple operation, and low toxicity. Therefore, at CD BioGlyco, we will utilize the action of microorganisms themselves to produce nucleotides.
Principle of nucleotide synthesis by microbial method: utilizing the fermentation action of microorganisms.
To produce nucleotides more efficiently, we will optimize the metabolic pathways and regulatory mechanisms of microorganisms, and also improve the permeability of cell membranes for nucleotides.
In addition to the production of nucleotides by direct microbial fermentation, we offer the following production pathways:
During the production of nucleotides by microorganisms, we continuously optimize the reaction process and provide multiple production paths for clients to choose from.
Fig.2 Three methods of microbial synthesis. (CD BioGlyco)
We select or engineer a microbial host based on the target nucleotide sugar. Using in silico metabolic modeling, we map key biosynthesis pathways and identify bottlenecks.
We apply CRISPR/Cas and base editing tools to modify the host genome: knocking out competing enzymes, overexpressing rate-limiting enzymes, and fine-tuning regulatory elements (e.g., ribosomal binding sites) to enhance flux.
Engineered microbes are cultured in scalable bioreactors, with parameters (pH, temperature, nutrient supply) optimized to maximize nucleotide sugar accumulation. Fed-batch strategies are used to extend production phases and minimize product degradation.
Nucleotide sugars are isolated from microbial lysates using a combination of ion-exchange chromatography and ultrafiltration, achieving. This step removes cellular contaminants, unreacted precursors, and byproducts.
DOI.: 10.1186/s12934-024-02452-8
Journal: Microbial Cell Factories
IF: 4.9
Published: 2024
Results: This study develops an engineered E. coli strain for enhanced guanosine production using combinatorial metabolic engineering. Researchers overexpressed Bacillus subtilis purine biosynthesis genes (Bspur operon) and PRPP synthase (prs), deleted guanosine catabolism genes (deoD, ppnP, gsk), and attenuated adenosine synthesis (purA). They redirected carbon flux by modifying EMP/ED pathways (deleting pfkA, edd, eda) and optimized redox cofactors. Transporter engineering (deleting nupG, overexpressing nepI) and strengthening guanosine synthesis (guaAB overexpression) further increased yield. The final strain achieved 289.8 mg/L guanosine in fed-batch fermentation – a 7-fold improvement over baseline, demonstrating efficient microbial nucleoside production.
Building on the foundation of our nucleotide microbial synthesis, which provides essential nucleotide precursors for glycosylation pathways, we seamlessly extend to advanced glyco-engineered cell construction services. These include targeted solutions such as the GPI anchor pathway-based, glycolipids (GSL) pathway-based, and N-linked glycan pathway-based glyco-engineered cell construction services, enabling precise manipulation of cellular glycosylation for diverse applications.
CD BioGlyco has a first-class laboratory with biosynthesis technology in line with international high-level technology. We have been researching in the direction of producing nucleotides for many years and our technology has become more mature. We provide custom production to each of our clients. If you are interested in our services, please feel free to contact us.
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