Biomimicry Promotes the Efficiency of a 10‐Step Sequential Enzymatic Reaction on Nanoparticles, Converting Glucose to Lactate

Abstract

For nanobiotechnology to achieve its potential, complex organic-inorganic systems must grow to utilize the sequential functions of multiple biological components. Critical challenges exist: immobilizing enzymes can block substrate-binding sites or prohibit conformational changes, substrate composition can interfere with activity, and multistep reactions risk diffusion of intermediates. As a result, the most complex tethered reaction reported involves only 3 enzymes. Inspired by the oriented immobilization of glycolytic enzymes on the fibrous sheath of mammalian sperm, here we show a complex reaction of 10 enzymes tethered to nanoparticles. Although individual enzyme efficiency was higher in solution, the efficacy of the 10-step pathway measured by conversion of glucose to lactate was significantly higher when tethered. To our knowledge, this is the most complex organic-inorganic system described, and it shows that tethered, multi-step biological pathways can be reconstituted in hybrid systems to carry out functions such as energy production or delivery of molecular cargo.

An Immobilized Enzyme Reactor for Spatiotemporal Control over Reaction Products

“…Recent work has demonstrated strategies to sequentially organize two or more enzymes on solid supports for performing multistep transformations . For example, packed‐bed reactors have been used to arrange enzymes in series using streptavidin, Ni‐NTA, and anti‐FLAG‐coated beads . Microfluidic devices integrated with valves or magnetic components have organized enzyme‐coated beads in well‐defined regions on a chip, which was beneficial for reducing dead volume .…”

Quantum Dots and Gold Nanoparticles as Scaffolds for Enzymatic Enhancement: Recent Advances and the Influence of Nanoparticle Size

“…Interestingly, going to larger sizes (microscale, >100 nm) can decrease the benefits from these types of scaffolds. One example is from the work of Mukai et al [84]. In this work, the authors used 500 nm Ni 2+ -NTA functionalized silica particles attached to the 10 enzymes of glycolysis (the 10 enzymes were split between three particles).…”

Expanding the boundary of biocatalysis: design and optimization of in vitro tandem catalytic reactions for biochemical production

“…However, in most cases, only 3 or fewer enzymes can be co-localized due to various challenges, such as the different responses of enzymes to different attachment methods (You et al, 2013; 2014). But recently Travis and coworkers demonstrated the possibility to create a 10-step enzymatic reaction on nanoparticles (Mukai et al, 2017). Although the activity of individual enzyme was higher in solution, the conversion of glucose to lactate was much higher when all the enzymes were tethered.…”