Integration of bioconversion and downstream processing: starch hydrolysis in an aqueous two-phase system

Integration of bioconversion and the first step(s) of down stream processing can be used as a means to increase the productivity of bioprocesses. This integration also gives the possibility to run the bioconversion in a continuous mode. We demonstrate the use of an aqueous two-phase system in combination with ultrafiltration to accomplish this. Conversion of native starch to glucose by alpha-amylase and glucoamylase was carried out in an aqueous two-phase system in connection with a membrane filtration unit. In this way, a continuous stream of glucose in buffer solution was obtained; the phase-forming polymers as well as the starch-degrading enzymes were recycled, and clogging of the ultrafiltration membrane was avoided. The process was carried out continuously in a mixer-settler reactor for a period of 8 days. The enzyme activities in the top and bottom phases and in the mixing chamber were monitored intermittently throughout the experiment. The optimum pH, temperature, and ionic strength for the activity of the enzyme mixture were determined. The settling time of phase systems containing varying amounts of PEG, crude dextran, and solid starch was studied. The activity and stability of enzyme mixtures was studied both in buffer medium and in the medium containing the polymers. The enzymes were found to be more active and stable in medium containing polymers than in the buffer solutions.