title: Enzyme Engineering for In Situ Immobilization.
abstract: Enzymes are used as biocatalysts in a vast range of industrial applications. Immobilization of enzymes to solid supports or their self-assembly into insoluble particles enhances their applicability by strongly improving properties such as stability in changing environments, re-usability and applicability in continuous biocatalytic processes. The possibility of co-immobilizing various functionally related enzymes involved in multistep synthesis, conversion or degradation reactions enables the design of multifunctional biocatalyst with enhanced performance compared to their soluble counterparts. This review provides a brief overview of up-to-date in vitro immobilization strategies while focusing on recent advances in enzyme engineering towards in situ self-assembly into insoluble particles. In situ self-assembly approaches include the bioengineering of bacteria to abundantly form enzymatically active inclusion bodies such as enzyme inclusions or enzyme-coated polyhydroxyalkanoate granules. These one-step production strategies for immobilized enzymes avoid prefabrication of the carrier as well as chemical cross-linking or attachment to a support material while the controlled oriented display strongly enhances the fraction of accessible catalytic sites and hence functional enzymes.
authors.affiliation: Institute of Fundamental Sciences, Massey University, Private Bag 11222, Palmerston North 4442, New Zealand. F.Rehm@massey.ac.nz.
authors.forename: Fabian B H
authors.initials: FB
authors.lastname: Rehm
authors.affiliation: Institute of Fundamental Sciences, Massey University, Private Bag 11222, Palmerston North 4442, New Zealand. S.X.Chen@massey.ac.nz.
authors.forename: Shuxiong
authors.initials: S
authors.lastname: Chen
authors.affiliation: Institute of Fundamental Sciences, Massey University, Private Bag 11222, Palmerston North 4442, New Zealand. B.Rehm@massey.ac.nz.
authors.forename: Bernd H A
authors.initials: BH
authors.lastname: Rehm
mesh_terms: D001419:Bacteria / Q000201:enzymology / Q000235:genetics; D055162:Biocatalysis; D004795:Enzyme Stability; D004798:Enzymes / Q000096:biosynthesis* / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D002479:Inclusion Bodies / Q000201:enzymology; D054813:Polyhydroxyalkanoates / Q000737:chemistry; D015202:Protein Engineering / Q000379:methods*
chemical_list: D004798:Enzymes; D004800:Enzymes, Immobilized; D054813:Polyhydroxyalkanoates
keywords: biocatalyst; enzyme; immobilization; inclusion bodies; protein particles; recombinant enzyme; self-assembly