Combining a Genetically Engineered Oxidase with Hydrogen-Bonded Organic Frameworks (HOFs) for Highly Efficient Biocomposites

Wied, P; Carraro, F; Bolivar, JM; Doonan, CJ; Falcaro, P; Nidetzky, B

Abstract

Enzymes incorporated into hydrogen-bonded organic frameworks (HOFs) via bottom-up synthesis are promising biocomposites for applications in catalysis and sensing. Here, we explored synthetic incorporation of d-amino acid oxidase (DAAO) with the metal-free tetraamidine/tetracarboxylate-based BioHOF-1 in water. N-terminal enzyme fusion with the positively charged module Z(basic2) strongly boosted the loading (2.5-fold; approximate to 500 mg enzyme g(material)(-1)) and the specific activity (6.5-fold; 23 U mg(-1)). The DAAO@BioHOF-1 composites showed superior activity with respect to every reported carrier for the same enzyme and excellent stability during catalyst recycling. Further, extension to other enzymes, including cytochrome P450 BM3 (used in the production of high-value oxyfunctionalized compounds), points to the versatility of genetic engineering as a strategy for the preparation of biohybrid systems with unprecedented properties.

Keywords: Biocatalysis; Hydrogen-Bonded Organic Frameworks; Immobilization; Metal-Organic Frameworks; Porous Carrier

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Hydrogen-bonded Organic Framework (HOF) Materials

Combining genetically engineered oxidases with hydrogen-bonded organic frameworks (HOFs) to produce biocomposites is an innovative material design approach that can be used in fields such as biocatalysis and biosensing. Genetic engineering technology is used to design and synthesize oxidases with specific catalytic activities. These oxidases may have efficient catalytic activity, stability, and specificity and can be used to catalyze various biotransformation reactions. Hydrogen bonded organic framework is a crystalline material formed by organic molecules through hydrogen bonding interactions. These frameworks have tunable pore structure and surface chemistry for applications such as adsorption, separation, and catalysis. Combining genetically engineered oxidases with hydrogen-bonded organic frameworks can achieve immobilization and stabilization of catalytic enzymes. The oxidase enzyme is firmly anchored in the material through surface or internal interactions with the hydrogen-bonded organic framework, maintaining its catalytic activity and specificity. At the same time, the pore structure and chemical properties of the hydrogen-bonded organic framework can adjust the rate and selectivity of the catalytic reaction. This biocomposite material has efficient catalytic performance and good stability and can be used in biocatalytic reactions, biosensors, bioenergy conversion and other fields, providing new possibilities for cross-research in biotechnology and materials science.