In this article we report on the characterization of the enzymatic synthesis of D-xylulose 5-phosphate using triosephosphate isomerase and transketolase. Two potential starting substrates are possible with this scheme. The data presented here allow a comparison of both routes for the synthesis, based on experimental information on reaction kinetics. Operational guidelines are proposed which should assist in the scale-up of such syntheses.

In this paper we report a method of semiquantitative process screening or route scouting developed to reduce the number of potential process flowsheets for a multistep biocatalytic conversion. Using the dual-enzyme-based synthesis of D-xylulose-5-phosphate as a model, a variety of possible processes was identified. Using data from a limited number of key experiments describing important process attributes, a screening procedure was proposed. Unattractive processes were eliminated early, and the best candidates were put forward as potential options for subsequent development. The method should prove applicable to a range of process synthesis problems.