Did you know that nucleotide triphosphates are key building blocks for RNA technologies, diagnostics and synthetic biology? acib offers a modular one-pot kinase cascade for the synthesis of all four canonical ribonucleoside triphosphates.
Background
Technology
acib has developed standardized one-pot enzymatic cascades for the production of ATP, CTP, GTP and UTP from the corresponding ribonu-cleosides. The platform combines substrate-specific nucleoside kinases with flexible yeast monophosphate kinases and acetate kinase-driven phosphate transfer. ScURA6 enables phosphorylation of AMP, CMP and UMP, while ScGMPK is used for GMP. E. coli acetate kinase drives the final phosphorylation step using acetyl phosphate as phosphate donor, with acetate as the by-product. The cascades were optimized to balance flux through all phosphorylation steps and suppress accumulation of NMP and NDP intermediates. Reactions reached full or near-full conversion (≥93%) at 50–100 mM substrate loading within 60–240 minutes. Preparative 10 mL syntheses delivered isolated NTP yields up to 89% and purities up to 99%. The platform can also be adapted toward noncanonical nucleosides and nucleoside analogues, offering a flexible starting point for tailored nucleotide synthesis.
Offer
acib offers to develop and optimize enzymatic NTP synthesis routes together with companies. Possible collaborations include feasibility studies, substrate screening, enzyme and reaction optimization, preparative synthesis, purification strategy development and adaptation to company-specific nucleotide targets.
IP developed in such projects can be fully transferred to the industrial partner.
Experts:
Dr. Martin Pfeiffer, Prof. Dr. Bernd Nidetzky
