Continuous production of oligoglucuronans by immobilized glucuronan lyase

A glucuronan lyase was incubated with sepharose matrices pre-activated with N-hydroxysuccinimide (NHS), cyanogen bromide (CNBr) or epoxy. The CNBr- and NHS-activated gels showed satisfactory immobilization yields whereas no enzyme could be immobilized using the epoxy coupling group. Glucuronan lyase immobilized on CNBr-gel ensured rapid conversion of several glucuronans into oligomers with an enzymatic activity identical to that of the free enzyme. As classically observed when using free enzymes, the acetylation degree of glucuronan limited enzyme activity. Nevertheless, this immobilized system made it easier to obtain accurately oligomers with different polymerization degrees, notably in modifying the glucuronans residence times in column. Thus oligoglucuronan pools with polymerization degrees between 2 and 25 could be obtained with a productivity ranging from 120 mg h−1 to 1.2 g h−1 using 0.9 ml of chromatographic gel with immobilized glucuronan lyase. This methodology opens the way to continuous and large oligoglucuronan productions.