Sequential Electrostatic Assembly of a Polymer Surfactant Corona Increases Activity of the Phosphotriesterase arPTE

Abstract

We present a new methodology for the generation of discrete molecularly dispersed enzyme− polymer−surfactant bioconjugates. Significantly, we demonstrate that >3-fold increase in the catalytic efficiency of the diffusion-limited phosphotriesterase arPTE can be achieved through sequential electrostatic addition of cationic and anionic polymer surfactants, respectively. Here, the polymer surfactants assemble on the surface of the enzyme via ion exchange to yield a compact corona. The observed rate enhancement is consistent with a mechanism whereby the polymer−surfactant corona gives rise to a decrease in the dielectric constant in the vicinity of the active site of the enzyme, accelerating the rate-determining product diffusion step. The facile methodology has significant potential for increasing the efficiency of enzymes and could therefore have a substantially positive impact for industrial enzymology.

Single Enzyme Nanoparticles with Improved Biocatalytic Activity through Protein Entrapment in a Surfactant Shell

“…In addition, modification of the acidic residues on the catalytic cleft may lead to enzymatic deactivation. To mitigate such challenges, Zhang et al prepared SENs via layer-by-layer assembly of oppositely charged surfactants on the protein surface . This appealing strategy improved enzymatic activity; however, stability may be insufficient at elevated ionic strengths when electrostatic interactions are weakened.…”

Solvent‐free liquid avidin as a step toward cold chain elimination

“…However, due to the surfactant corona, there was a reduction in binding site availability for [C‐Av][S], indicating a potential trade‐off between high thermal stability and binding activity. To increase the activity of solvent‐free biofluids, there are possible methodologies emerging from the literature, such as the use of cationic and anionic surfactants instead of cationization with a diamine (Zhang et al, 2019), and the use of site‐directed mutagenesis (Zhou et al, 2019).…”

“…The reversibility of the binding pocket was investigated by displacing HABA (λ max = 350 nm) from the Av-HABA complex (λ max = 500 nm) with biotin. The absolute data showed that Av possessed a K d of 1.12 × 10 −5 M (Table S6) as the use of cationic and anionic surfactants instead of cationization with a diamine (Zhang et al, 2019), and the use of site-directed mutagenesis (Zhou et al, 2019).…”

Preparation and application of solvent-free liquid proteins with enhanced thermal and anhydrous stabilities

“…Pérez et al later showed that it was possible to improve on the activity through careful consideration of the cationization process, doubling the activity of the solvent-free lipase as a result. 66 Recently, solvent-free enzyme substrate scope has been expanded with Zhang et al demonstrating phosphotriesterase activity 67 and Mukhopadhayay et al demonstrating protease activity. 68 In addition, Atkins et al…”

“…The first is Please do not adjust margins Please do not adjust margins based on controlling the chemistry of the modification: Deller et al sought to control the kinetics of the cationization process to ensure active site preservation in thrombin 73 , and Zhang et al used a combination of anionic and cationic polymer surfactants to circumvent the requirement of covalent modification entirely. 67 The second approach is realized through utilisation of synthetic biology to modify the protein itself to be more amenable to chemical modification. For example, Zhou et al used site directed mutagenesis to ensure that chemical modification did not alter the active site of lipase, increasing subsequent activity as a result.…”