Nucleoside Cell Factory Synthesis Service
The fusion of a nitrogenous base with a sugar moiety, usually ribose or deoxyribose, is used in the production of nucleosides. This fusion can occur spontaneously within cells during nucleotide biosynthesis or in a laboratory. Enzymes aid in the synthesis of nucleosides by catalyzing the glycosidic connection between the nitrogenous base and the sugar. Chemical procedures can be used in the laboratory to synthesize nucleosides. Uridine, a pyrimidine nucleoside, is widely used in the pharmaceutical industry. A recent research emphasis is on uracil cell factory manufacturing, with CRISPR/Cas9-mediated chromosomal integration functioning as an effective technique for developing appropriate cell factories for this purpose.
We leverage metabolic engineering of recombinant bacteria equipped with high-efficiency nucleoside-synthesizing enzymes, utilizing CRISPR/Cas9-mediated chromosomal integration for precise strain development. Our technology optimizes growth conditions and metabolic pathways to enable scalable, sustainable nucleoside production for pharmaceutical applications, minimizing environmental impact while ensuring high yields.
Nucleosides and nucleoside derivatives, integral components of antiviral and anticancer medications, are often synthesized using Chemical Methods. However, this approach involves hazardous compounds, leading to environmental harm. Enzymatic synthesis, like the production of 5-methyluridine, is an alternative, but bacterial cell requirements limit its scalability. Cell factories offer a novel avenue for nucleoside manufacturing. Leveraging our robust experimental platform, CD BioGlyco employs recombinant bacteria with potent nucleoside-producing enzymes for cell factory-based nucleoside and analog production. Our services encompass the Technology Development Phase and scalable production, ensuring clients' needs are met. We also extend Nucleotide Synthesis Services, including Chemical Synthesis, Microbial Synthesis, and Enzymatic Digestion, ensuring comprehensive support for diverse research and industrial applications.
It is critical to emphasize that the success of nucleoside synthesis utilizing a cell factory is dependent on meticulous strain engineering, growth environment optimization, and efficient metabolic pathway design. Furthermore, sophisticated techniques like CRISPR/Cas9-Mediated Genome Editing can aid in the construction of high-yielding cell factories for nucleoside manufacturing.
Our nucleoside cell factory synthesis service follows a systematic and optimized workflow designed for efficiency and high-yield production:
We begin by selecting a suitable microbial host, often Escherichia coli, due to its well-understood genetics and robust growth characteristics. Initial engineering focuses on introducing essential biosynthetic pathways, such as integrating multi-gene operons into the host chromosome to establish foundational nucleoside synthesis capabilities.
A crucial step involves identifying and knocking out genes responsible for the degradation of the target nucleoside monomer. This prevents the breakdown of the synthesized product and significantly enhances its accumulation.
We then focus on enhancing the cellular supply of key metabolic precursors. This involves genetic modifications such as overexpressing rate-limiting enzymes (e.g., CPS) and deleting genes that compete for these precursors (e.g., argF, iclR, thrA). This strategy ensures a robust flow of building blocks towards the desired nucleoside synthesis pathway.
To maximize the extracellular yield, we modify or delete genes encoding nucleoside transport proteins to facilitate the efficient secretion of the synthesized nucleoside monomers from the cell into the culture medium.
The engineered strains undergo rigorous fermentation optimization in controlled bioreactor environments. This involves fine-tuning culture conditions such as media composition, pH, temperature, and aeration rates to achieve optimal growth and maximum product titer and productivity in fed-batch fermentation.
DOI.: 10.1186/s12934-024-02452-8
Journal: Microbial Cell Factories
IF: 4.9
Published: 2024
Results: This study describes the use of combinatorial metabolic engineering in Escherichia coli MG1655 to significantly increase guanosine production. The researchers first chromosomally integrated Bacillus subtilis's purine synthesis operon (Bspur) and overexpressed phosphoribosyl pyrophosphate synthase (prs), while knocking out degradation genes (deoD, ppnP, gsk). They then attenuated the adenosine branch pathway (modified purA), eliminated feedback inhibition by deleting purR, and redirected metabolic flux by disrupting glycolysis/ED pathways (pfkA, edd, eda knockout with glpX overexpression) while rebalancing redox cofactors. Finally, transporter engineering (nupG deletion + nepI overexpression) and strengthening the guanosine synthesis pathway (overexpressing guaAB) further boosted production. The optimized strain MQ39 achieved a guanosine titer of 289.8 mg/L in shake-flask fed-batch fermentation after 72 hours, demonstrating the effectiveness of this systematic approach.
Beyond precision nucleoside synthesis via our nucleoside cell factory synthesis, we empower next-stage functionalization through specialized Glycogene Engineering Service. Harnessing these enzymatic tools unlocks tailored glycoconjugate synthesis:
Glucosyltransferase Engineering Service
Build glucose-linked nucleosides for targeted oligosaccharide/prodrug applications.
Galactosyltransferase Engineering Service
Design galactosyl-adducted compounds oligosaccharide/prodrug applications.
Mannosyltransferase Engineering Service
Engineer mannose-functionalized analogs for vaccine adjuvants and lectin-targeted delivery.
CD BioGlyco provides cell factory synthesis services for synthesizing nucleosides, and we have a high-level lab to process high-quality synthesis experiments. Please feel free to contact us for more detailed information.
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