Due to their efficiency, selectivity, and environmental sustainability, there are significant opportunities for enzymes in chemical synthesis and biotechnology. However, as the three-dimensional active structure of enzymes is predominantly maintained by weaker non-covalent interactions, thermal, pH and chemical stressors can modify or eliminate activity. Metal-organic Frameworks (MOFs), which are extended porous network materials assembled by a bottom-up building block approach from metal-based nodes and organic linkers, can be used to afford protection to enzymes. The self-assembled structures of MOFs can be used to encase an enzyme in a process called encapsulation when the MOF is synthesized in the presence of the biomolecule. Alternatively, enzymes can be infiltrated into mesoporous MOF structures or surface bound via covalent or non-covalent processes. Integration of MOF materials and enzymes in this way affords protection and allows the enzyme to maintain activity in challenge conditions (e.g. denaturing agents, elevated temperature, non-native pH and organic solvents). In addition to forming simple enzyme/MOF biocomposites, other materials can be introduced to the composites to improve recovery or facilitate advanced applications in sensing and fuel cell technology. This review canvasses enzyme protection via encapsulation, pore infiltration and surface adsorption and summarizes strategies to form multi-component composites. Also, given that enzyme/MOF biocomposites straddle materials chemistry and enzymology, this review provides an assessment of the characterization methodologies used for MOF-immobilized enzymes and identifies some key parameters to facilitate development of the field. CONTENTSMOF-based enzyme biocomposite compositions and the concept of encapsulation 3.MOF-based enzyme biocomposites formed via encapsulation 3.1.Templating methods 3.2.One-pot embedding (non-templated) 3.3.Parameters influencing the chemistry of enzyme@MOF biocomposites 3.3.1. Additives 3.3.2.Enzyme suface chemistry 3.3.3.MOF precusors and structures 3.4.Alternative synthesis strategies 4.Infiltration (post insertion of enzymes in preformed MOFs) 4.1.Early results and the advantages of MOFs for infiltration 4.2.Tuning the framework structure in infiltrated enzyme@MOF biocomposites 4.3.Towards applications of infiltrated enzyme@MOFs 5.Surface bound enzymes 5.1. Immobilization via physical adsorption 5.2. Immobilization via coordinate bonds 5.3. Immobilization via covalent bonding 5.4. Enzymatic activity upon surface-immobilization 6. Multicomponent biocomposites 7. Characterization of MOF immobilized enzymes 7.1. Overview 7.2. Key immobilization parameters 7.3. One-pot enzyme MOF formation 7.4. Highlights of MOF-immobilized enzyme performance 7.4.1. Experimental determination of key immobilization parameters 7.4.1.1. Determination of protein concentrations 7.4.1.2. Activity determination 7.4.2. Advanced characterization of immobilized enzymes 7.4.2.1. Apparent enzyme kinetics 7.4.2.2. Structural analysis and localization o...

A porous model bioreactor is obtained combining zeolitic imidazolate framework ZIF-8 with horseradish peroxidase and iron oxide magnetic nanoparticles in a one-pot process, in water at room temperature.

In this study, we coupled a well‐established whole‐cell system based on E. coli via light‐harvesting complexes to Rieske oxygenase (RO)‐catalyzed hydroxylations in vivo. Although these enzymes represent very promising biocatalysts, their practical applicability is hampered by their dependency on NAD(P)H as well as their multicomponent nature and intrinsic instability in cell‐free systems. In order to explore the boundaries of E. coli as chassis for artificial photosynthesis, and due to the reported instability of ROs, we used these challenging enzymes as a model system. The light‐driven approach relies on light‐harvesting complexes such as eosin Y, 5(6)‐carboxyeosin, and rose bengal and sacrificial electron donors (EDTA, MOPS, and MES) that were easily taken up by the cells. The obtained product formations of up to 1.3 g L−1 and rates of up to 1.6 mm h−1 demonstrate that this is a comparable approach to typical whole‐cell transformations in E. coli. The applicability of this photocatalytic synthesis has been demonstrated and represents the first example of a photoinduced RO system.

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 Zbasic2 strongly boosted the loading (2.5‐fold; ≈500 mg enzyme gmaterial−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.