We report herein the use of nanofibrillated cellulose (NFC) for development of enzyme assemblies in an oriented manner for biotransformation with in situ cofactor regeneration. This is achieved by developing fusion protein enzymes with cellulose-specific binding domains. Specifically, lactate dehydrogenase and NADH oxidase were fused with a cellulose binding domain, which enabled both enzyme recovery and assembling in essentially one single step by using NFC. Results showed that the binding capacity of the enzymes was as high as 0.9 μmol-enzyme/g-NFC. Compared to native parent free enzymes, NFC-enzyme assemblies improved the catalytic efficiency of the coupled reaction system by over 100%. The lifetime of enzymes was also improved by as high as 27 folds. The work demonstrates promising potential of using biocompatible and environmentally benign bio-based nanomaterials for construction of efficient catalysts for intensified bioprocessing and biotransformation applications.
Keywords: Cellulose binding domain; Cofactor regeneration; Lactate dehydrogenase; NADH oxidase; Nanocellulose.