Abstract
Chondroitin sulfates (CS) are linear glycosaminoglycans that have important applications in the medical and food industries. Engineering bacteria for microbial production of CS will facilitate the one?step, scalable production with good control over sulfation levels and positions in contrast to extraction from animal sources. Advancing towards this goal, we engineered E. coli to accumulate 3’?phosphoadenosine?5’?phosphosulfate (PAPS), the universal sulfate donor, using traditional metabolic engineering approaches. PAPS is one of the least explored components required for biosynthesis of CS.
The resulting engineered E. coli strain shows a ~1000?fold increase in intracellular PAPS concentrations. This study also reports, for the first time, in vitro biotransformation of CS using PAPS, chondroitin and chondroitin?4?sulfotransferase (C4ST) all synthesized from different engineered E. coli strains made in our laboratory. We show a 10.4?fold increase in the amount of CS produced by biotransformation by employing PAPS from the engineered PAPS?accumulating strain. The data from the biotransformation experiments also help us evaluate reaction components that need improved production to achieve a one? step microbial synthesis of CS. This would provide a new platform to produce CS.
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