Hierarchically Engineered Multi‐Enzyme Nanoreactors for in vitro Drug Biosynthesis and Pathway Transplantation Into Cells
Abstract
ABSTRACT Even though most proteins evolved to function within multi‐protein systems such as metabolic networks or signaling pathways, most technical protein applications are based on isolated proteins, for example therapeutic antibodies or industrial enzymes. Reliable methods for the stabilization, in vitro operation and intracellular delivery of multi‐protein systems could unlock new applications in green biotechnology, diagnostics, and medical therapy. We demonstrate that the entire violacein biosynthesis pathway, consisting of up to six separately purified enzymes, can be infiltrated into a hierarchically etched MIL‐101(Fe) (eMIL) metal organic framework. eMIL nanoreactors reshaped pathway dynamics and reaction flows, multiplied violacein yield in vitro and enabled pathway reuse, lyophilization and storage. Moreover, eMIL nanoreactors delivered the six‐protein system into mammalian cells, where it produced violacein from cell‐provided substrates and cofactors. These findings pave the way for the design of “smart” stimuli‐responsive multi‐enzyme nanoreactors for biotechnological and medical applications.
Article Details
Authors (13)
Ainur Sharip
Biomedical Sciences Division King Abdullah University of Science and Technology (KAUST) Thuwal Saudi Arabia
Somayah S. Qutub
Biomedical Sciences Division King Abdullah University of Science and Technology (KAUST) Thuwal Saudi Arabia
Manar M. Farooqui
Biomedical Sciences Division King Abdullah University of Science and Technology (KAUST) Thuwal Saudi Arabia
Walaa Baslyman
Physical Science and Engineering Division (PSE) King Abdullah University of Science and Technology (KAUST) Thuwal Saudi Arabia
Nida Khalfay
Biomedical Sciences Division King Abdullah University of Science and Technology (KAUST) Thuwal Saudi Arabia
Lukman O. Alimi
Smart Hybrid Materials (SHMs) Laboratory, Physical Science and Engineering (PSE) Division
Patricia Lopez Sanchez
Analytical Chemistry Core Laboratory KAUST Thuwal Saudi Arabia
Lingyun Zhao
Imaging and Characterization Core Lab, King Abdullah University of Science and Technology
Milena Chernyshevskaia
Biomedical Sciences Division King Abdullah University of Science and Technology (KAUST) Thuwal Saudi Arabia
Giovanni Colombo
Biomedical Sciences Division King Abdullah University of Science and Technology (KAUST) Thuwal Saudi Arabia
Niveen M. Khashab
Smart Hybrid Materials (SHMs) Laboratory, Physical Science and Engineering (PSE) Division
Stefan T. Arold
Raik Grünberg