Enzyme Nanocomplexes With Polymer Shells

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Solution Overview

Problem

Current enzyme-based applications are limited to single enzymes or their mixtures, and existing methods for constructing multiple-enzyme architectures often result in decreased enzyme specificity and activity, with challenges in designing suitable host cells and spacers, and lack general applicability.

Innovation Solution

The development of artificial enzyme nanocomplexes comprising multiple enzymes anchored to a polymeric network, allowing for cooperative enzymatic functions and enhanced stability, with a permeable shell that facilitates specific reactions and reduces toxic intermediate release.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Stability of the object's composition

If multiple enzymes are randomly immobilized within inorganic and polymeric materials in the forms of thin films or large particles, then the structural stability is improved, but the enzyme specificity and activity are decreased

Engineering Contradiction:
Improvestructural stabilityVSAvoidenzyme specificity and activity
Core Design Contradiction:
Stability of the object's compositionVSReliability

Solution Approach 1:

The invention divides the enzyme system into discrete nanoscale compartments (nanocomplexes) rather than using continuous films or large particles. Each nanocomplex contains a defined number of enzyme molecules organized in specific spatial arrangements, segmenting the enzyme population into functional units that maintain both stability and activity. This segmentation allows enzymes to retain their native-like environment while gaining structural support.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention uses flexible polymer shells to encapsulate enzyme molecules, creating a protective yet permeable environment. The polymer shell provides structural stability while its flexible nature and controlled porosity allow substrates and products to diffuse freely, maintaining enzyme activity and specificity. This approach contrasts with rigid inorganic materials that may restrict enzyme movement and substrate access.

Inventive Principle:
Principle #30Flexible shells and thin films

2Adaptability or versatility

If fusion-protein techniques are used to construct multiple-enzyme architectures, then the designability is improved, but the enzyme specificity and activity are lost or decreased

Engineering Contradiction:
ImprovedesignabilityVSAvoidenzyme specificity and activity
Core Design Contradiction:
Adaptability or versatilityVSReliability

Solution Approach 1:

The invention introduces polymer molecules as intermediaries between enzyme molecules, replacing direct protein-protein fusion. The polymer acts as a spacer and organizer that brings enzymes into close proximity without forming covalent fusion proteins. This intermediary approach allows enzymes to maintain their native structures and active sites while achieving the desired spatial organization for multi-enzyme functionality.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention creates composite nanocomplexes consisting of enzyme molecules combined with synthetic or biological polymer materials. This composite structure combines the catalytic functionality of enzymes with the structural properties and design flexibility of polymers, achieving both high designability and preserved enzyme activity through non-covalent associations rather than fusion protein construction.

Inventive Principle:
Principle #40Composite materials

3Manufacturing precision

If single proteins are encapsulated into nanogels, then the size uniformity is improved, but the cooperative enzymatic functions are not achieved

Engineering Contradiction:
Improvesize uniformityVSAvoidcooperative enzymatic functions
Core Design Contradiction:
Manufacturing precisionVSProductivity

Solution Approach 1:

The invention merges multiple enzyme molecules within a single nanocomplex structure, enabling cooperative catalysis. By combining several different enzymes in defined stoichiometric ratios within uniformly sized nanocomplexes, the system achieves both size precision and cooperative functionality. The enzymes work together in sequence or parallel to catalyze multi-step reactions, enhancing overall productivity beyond what single enzymes can achieve.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The invention nests multiple enzyme molecules within a shared polymer shell, creating a hierarchical structure where individual enzymes are contained within a common protective compartment. This nesting arrangement maintains size uniformity of the overall nanocomplex while allowing enzymes to be positioned in optimal orientations for cooperative catalysis, with substrates diffusing through the polymer shell to access multiple enzymes in sequence.

Inventive Principle:
Principle #7Nested doll (Nesting)

4Productivity

If toxic intermediates are generated during metabolic processes, then the reaction productivity is improved, but the cellular damage is increased

Engineering Contradiction:
Improvereaction productivityVSAvoidcellular damage from toxic intermediates
Core Design Contradiction:
ProductivityVSObject-affected harmful factors

Solution Approach 1:

The invention converts the harmful effect of toxic intermediates into a beneficial outcome by co-locating detoxifying enzymes within the same nanocomplex. Toxic intermediates generated by one enzyme are immediately captured and converted by neighboring enzymes in the same compartment, transforming a harmful byproduct into a substrate for the next catalytic step. This approach maintains high reaction productivity while eliminating cellular toxicity through integrated detoxification pathways.

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The enzyme nanocomplexes exhibit enhanced catalytic efficiency and stability, enabling effective breakdown of toxic compounds like hydrogen peroxide while minimizing intermediate toxicity, with improved designability and applicability across various environments.

Implementation Method 1

a polymeric network which is anchored to at least one of the at least two different enzymes

Methodology Applied
Scientific EffectCovalent bonding: Chemical Bonding

Implementation Method 2

the enzymes cooperatively carry out their enzymatic functions

Methodology Applied
Scientific EffectEnzyme catalysis: Catalysis

Implementation Method 3

a permeable shell that facilitates specific reactions and reduces toxic intermediate release

Methodology Applied
Scientific EffectDiffusion: Diffusion

Data Source

PatentEP2729569B1Multiple-enzyme nanocomplexes
Publication Date: 2019.12.11 RGT UNIV OF CALIFORNIA
  • EP2729569B1 patent drawingFigure 1
  • EP2729569B1 patent drawingFigure 2a~2h
  • EP2729569B1 patent drawingFigure 3a~3dii

AI summary

Provided are nanocomplexes having at least two different enzymes and a polymeric network anchored to at least one of the enzymes. In some embodiments, the activities of the enzymes catalyze a cascade reaction.