Gram‐Scale Production of GDP‐β‐ <scp>l‐</scp> fucose with Multi‐Enzyme Cascades in a Repetitive‐Batch Mode

Guanosine 5’-diphospho-β-l-fucose (GDP-Fuc) is an important and expensive nucleotide sugar only available in low quantities. We established two processes for the gram-scale production of GDP-Fuc with multi-enzyme cascades. used bifunctional enzyme fucokinase/GDP-fucose pyrophosphorylase from Bacteroides fragilis either ATP-regeneration system using polyphosphate or excess ATP. The optimized modules were scaled up to a 20 mL repetitive-batch 15 12 repetitive batches ATP excess, respectively. With final product amount 1.1 g (31 % yield) was obtained. process reached 1.6 (57 yield). Exceptional high total turnover numbers 27 gP*gE−1 31 gP*gE−1, as well space-time-yields 0.083 gp*L−1*h−1 0.217 each Furthermore, we purification protocol that enables 95 removal free phosphates solution, ease further purification.