Controlled Orientation of Active Sites in a Nanostructured Multienzyme Complex
Autor: | Sung In Lim, Inchan Kwon, Younghan Jung, Jaehyun Cha, Eun Sil Choi, Jinhwan Cho, Byungseop Yang, Yong Hwan Kim |
---|---|
Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
010402 general chemistry Formate dehydrogenase Pseudomonas fluorescens 01 natural sciences Article 03 medical and health sciences chemistry.chemical_compound Cascade reaction Multienzyme Complexes Catalytic Domain Mannitol Binding site Probability chemistry.chemical_classification Mannitol Dehydrogenases Multidisciplinary Binding Sites biology Active site DNA Thiobacillus Formate Dehydrogenases 0104 chemical sciences Amino acid Nanostructures Oxygen Kinetics 030104 developmental biology Mannitol dehydrogenase chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Methanocaldococcus biology.protein Biophysics Bioorthogonal chemistry |
Zdroj: | Scientific Reports SCIENTIFIC REPORTS(6) |
ISSN: | 2045-2322 |
Popis: | Multistep cascade reactions in nature maximize reaction efficiency by co-assembling related enzymes. Such organization facilitates the processing of intermediates by downstream enzymes. Previously, the studies on multienzyme nanocomplexes assembled on DNA scaffolds demonstrated that closer interenzyme distance enhances the overall reaction efficiency. However, it remains unknown how the active site orientation controlled at nanoscale can have an effect on multienzyme reaction. Here, we show that controlled alignment of active sites promotes the multienzyme reaction efficiency. By genetic incorporation of a non-natural amino acid and two compatible bioorthogonal chemistries, we conjugated mannitol dehydrogenase to formate dehydrogenase with the defined active site arrangement with the residue-level accuracy. The study revealed that the multienzyme complex with the active sites directed towards each other exhibits four-fold higher relative efficiency enhancement in the cascade reaction and produces 60% more D-mannitol than the other complex with active sites directed away from each other. |
Databáze: | OpenAIRE |
Externí odkaz: |