Coflobiopro: Continuous flow biocatalytic process for production of value-added chemicals using magnetic nanoparticle immobilized enzyme microreactors

The Coflobiopro project will establish an optimally functioning continuous enzyme cascade with pure glycerol as substrate to produce acetoin and 2,3-butanediol(2,3BD) as the major products. 2,3-BD has an extremely high industrial value with potential application in the sectors of aviation fuel, rubber, pharmaceuticals, methyl-ethyl ketone (fuel additive), foodstuff(diacetyl) and acetoin production (volatile food additive). The current global market values of the major downstream products from 2,3-BD is reported to be $43 billion per year with the value of acetoin reported to be approximately $30000 per ton. The project will utilize currently prevailing technological advancements in a catalytic flow through system packed in a single microreactor platform. Effective research strategies have been put forth towards generation of 2, 3 butanediol (2,3-BD) by bioengineered microbial species from renewable substrates but with limited success rate since the terminal product is low in volume. To counter this, we have devised a synthetic metabolic pathway mediated by magnetic nanoparticle (MNP) immobilized biocatalysts in a packed bed enzymatic microreactor. Initially, we will ascertain the optimal conditions for product outputs for each of the sequential reaction steps in the study. The individual reactions catalyzed by the MNP immobilized enzymes alditol oxidase, dihydroxy acid dehydratase, a-acetolactate synthase, a-acetolactate decarboxylase and butanediol dehydrogenase in individual reactors will determine the conditions that lends optimal output of the resultant products. Finally, the optimized proportions of the MNP-immobilised enzymes will be loaded in a single reactor and the reactions will be performed under optimized conditions in continuous mode. The novelty of Coflobiopro is that it incorporates optimally functioning MNP immobilized enzymes aligned in a sequential manner in a microreactor for generation of
value-added biofuels in a continuous biocatalytic cascade.