- Autor
- Zhong, Chao
- Duić, Božidar
- Bolivar, Juan Manuel
- Nidetzky, Bernd
- TitelThree-Enzyme Phosphorylase Cascade Immobilized on Solid Support for Biocatalytic Synthesis of Cello oligosaccharides
- Datei
- DOI10.1002/cctc.201901964
- Erschienen inChemCatChem
- Band12
- Erscheinungsjahr2020
- Heft5
- Seiten1350-1358
- LicenceCC BY 4.0
- ISSN1867-3899
- Download Statistik69
- Peer ReviewJa
- AbstractEnzyme cascades are promising for multistep biocatalytic syn- thesis, but their effective use beyond the proof-of-concept stage is challenging. Strategies to recycle the individual enzymes are critical for the applicability of such cascades. Immobilization on solid support is well developed for single enzymes but remains difficult for enzyme ensembles. Here, we show a controlled co-immobilization of three glycoside phos- phorylases to establish a highly active and recyclable biocatalyst for the conversion of sucrose and glucose into soluble (short- chain) cello oligosaccharides. We use protein fusion with the binding module Zbasic2 to enable non-covalent surface tethering of all enzymes according to a uniform principle and in a programmable fashion. We thus achieve loading of the phosphorylases in an activity ratio optimal for the overall conversion and for controlling the cello oligosaccharide chain length (� 6), hence the solubility, in the reaction. We demon- strate efficient production of ~ 12 g/L cello oligosaccharides with integrated enzyme re-use, retaining ~ 85 % of the overall initial activity after five reaction cycles. This study presents a major advance toward the practical use of systems bio-catalysis on solid support.
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