Molecular crowding elicits the acceleration of enzymatic crosslinking of macromolecular substrates

Sato, Ryo; Minamihata, Kosuke; Wakabayashi, Rie; Goto, Masahiro; Kamiya, Noriho

doi:10.1039/d2ob01549h

Cited by 5 publications

(4 citation statements)

References 40 publications

(49 reference statements)

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“…Changes in the dielectric constant can occur in densely packed assemblies of proteins 20 , 21 , which could lead to changes in the electrostatic interactions between the substrate and the active site of the enzyme. The underlying mechanisms of this phenomenon remain to be elucidated, such as the influence of physicochemical parameters within the enzyme assembly on the enzymatic reaction mechanism, for instance, macromolecular crowding 22 , 23 , pH 24 , 25 or relative permittivity 20 , 21 . In addition, since the enzyme used in this study forms multimers (Table 1 ), changes in multimers during the formation of enzyme assemblies may be important changes in k cat or K M .…”

Section: Discussionmentioning

confidence: 99%

Activation of oxidoreductases by the formation of enzyme assembly

Ura,

Sakakibara,

Hirano

et al. 2023

Sci Rep

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Biological properties of protein molecules depend on their interaction with other molecules, and enzymes are no exception. Enzyme activities are controlled by their interaction with other molecules in living cells. Enzyme activation and their catalytic properties in the presence of different types of polymers have been studied in vitro, although these studies are restricted to only a few enzymes. In this study, we show that addition of poly-l-lysine (PLL) can increase the enzymatic activity of multiple oxidoreductases through formation of enzyme assemblies. Oxidoreductases with an overall negative charge, such as l-lactate oxidase, d-lactate dehydrogenase, pyruvate oxidase, and acetaldehyde dehydrogenase, each formed assemblies with the positively charged PLL via electrostatic interactions. The enzyme activities of these oxidoreductases in the enzyme assemblies were several-folds higher than those of the enzyme in their natural dispersed state. In the presence of PLL, the turnover number (kcat) improved for all enzymes, whereas the decrease in Michaelis constant (KM) was enzyme dependent. This type of enzyme function regulation through the formation of assemblies via simple addition of polymers has potential for diverse applications, including various industrial and research purposes.

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Section: Discussionmentioning

confidence: 99%

Activation of oxidoreductases by the formation of enzyme assembly

Ura,

Sakakibara,

Hirano

et al. 2023

Sci Rep

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“…The preparation and use of the proteins MTG, LysM-Q, and LysM-muGFP-Q were performed as described previously 13,23 .…”

Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

“…The preparation and use of the proteins MTG, LysM-Q, and LysM-muGFP-Q were performed as described previously. 13,23 Fig. 4 Cell viability of Hek293T cells (5000 cells per well) incubated with LysM-lipids alone (a) and in combination of AmB (b).…”

Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

Exploring the molecular structure of lipids in the design of artificial lipidated antifungal proteins

Saputra,

Safaat,

Uchida

et al. 2024

RSC Pharm.

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“…Thus, for the last many decades, efforts have been made to improve lipase activity in different micro-heterogeneous domains like reverse micelles, microemulsions, [33] hydrogels, [34] polymers, and others. [35] In such self-aggregated structures, water-insoluble substrates can reach the active site of lipase at the water-oil interface. However, in most of these methods, lipase loses its native structure and conformation to a notable extent due to the exposure of the enzyme to the non-aqueous environment.…”

Section: Introductionmentioning

confidence: 99%

Peptide Amphiphilic Supramolecular Nanogels: Competent Host for Notably Efficient Lipase‐Catalyzed Hydrolysis of Water‐Insoluble Substrates

et al. 2023

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The present work depicts the development of stable nanogels in an aqueous medium that were exploited for efficient surface‐active lipase‐catalyzed hydrolysis of water‐insoluble substrates. Surfactant‐coated gel nanoparticles (neutral NG1, anionic NG2, and cationic NG3) were prepared from peptide amphiphilic hydrogelator (G1, G2, and G3, respectively) at different hydrophilic and lipophilic balance (HLB). Chromobacterium viscosum (CV) lipase activity towards hydrolysis of water‐insoluble substrates (p‐nitrophyenyl‐n‐alkanoates (C4–C10)) in the presence of nanogels got remarkably improved by ~1.7–8.0 fold in comparison to that in aqueous buffer and other self‐aggregates. An increase in hydrophobicity of the substrate led to a notable improvement in lipase activity in the hydrophilic domain (HLB>8.0) of nanogels. The micro‐heterogeneous interface of small‐sized (10–65 nm) nanogel was found to be an appropriate scaffold for immobilizing surface‐active lipase to exhibit superior catalytic efficiency. Concurrently, the flexible conformation of lipase immobilized in nanogels was reflected in its secondary structure having the highest α‐helix content from the circular dichroism spectra.

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Molecular crowding elicits the acceleration of enzymatic crosslinking of macromolecular substrates

Abstract: Cytoplasm contains high concentrations of biomacromolecules. Protein behavior under such crowded conditions is reportedly different from that in an aqueous buffer solution, mainly owing to the effect of volume exclusion...

Cited by 5 publications

References 40 publications

Activation of oxidoreductases by the formation of enzyme assembly

Activation of oxidoreductases by the formation of enzyme assembly

Exploring the molecular structure of lipids in the design of artificial lipidated antifungal proteins

Peptide Amphiphilic Supramolecular Nanogels: Competent Host for Notably Efficient Lipase‐Catalyzed Hydrolysis of Water‐Insoluble Substrates

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