Enzyme Induction Using Amide Amino Acids

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Solution Overview

Problem

Current methods for inducing and stabilizing enzyme activity in microorganisms, particularly those capable of degrading nitrile-containing compounds, face challenges such as the need for hazardous chemicals and instability of enzymes, limiting their practical application in commercial processes.

Innovation Solution

The use of amide-containing amino acids like asparagine and glutamine as inducers in the growth media for microorganisms, combined with immobilization techniques, to enhance enzymatic activity and stability without requiring hazardous nitriles, thereby stabilizing enzymes like nitrile hydratase and asparaginase I for extended periods.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If hazardous nitriles are used to induce enzyme activity in microorganisms, then enzyme production is enhanced, but safety and environmental hazards increase

Engineering Contradiction:

Improveenzyme productionVSAvoidhazardous chemicals

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent uses amide-containing amino acids (asparagine, glutamine) as intermediary substances to induce enzyme activity. These amino acids serve as safe mediators that trigger the same enzymatic response as hazardous nitriles would, but without the associated safety and environmental risks. The amino acids are metabolized by the microorganisms to produce the desired enzymatic activity.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent employs readily available, non-hazardous amide-containing amino acids as induction agents. These substances are inexpensive, easily decomposed by microorganisms, and leave no harmful residues, replacing the need for persistent hazardous nitrile compounds while maintaining effective enzyme induction.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

2Reliability

If traditional enzyme stabilization methods are used, then enzyme activity is maintained, but enzyme stability is insufficient for extended periods

Engineering Contradiction:

Improveenzyme stabilityVSAvoiduseful life of enzymatic activity

Core Design Contradiction:

ReliabilityVSDuration of action of stationary object

Solution Approach 1:

The patent applies preliminary induction using amide-containing amino acids during the microorganism growth phase to pre-activate and stabilize enzyme production. This preliminary action ensures that enzymes are produced at optimal levels before the actual application, extending their useful life and maintaining stability over extended periods.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent changes the chemical parameters of the growth medium by incorporating specific amide-containing amino acids, which alter the metabolic state of microorganisms and lead to enhanced enzyme stability. This parameter change in the culture conditions results in enzymes that maintain activity for longer durations.

Inventive Principle:

Principle #35Parameter changes

3Reliability

If immobilization techniques are applied to stabilize enzymes, then enzyme reusability is improved, but process complexity increases

Engineering Contradiction:

Improveenzyme reusabilityVSAvoidimmobilization process

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent enables microorganisms to self-produce and self-stabilize enzymes through metabolic processing of amide-containing amino acids. The microorganisms naturally incorporate these amino acids into enzyme structures, providing self-stabilization without requiring complex external immobilization systems, thereby maintaining simplicity while achieving reusability.

Inventive Principle:

Principle #25Self-service

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

This approach allows for the induction and stabilization of enzymes at higher levels, enabling effective detoxification of nitrile-containing waste streams and extending the useful life of enzymatic activity beyond typical stability limits without using hazardous chemicals, making the process safer and more efficient.

Implementation Method 1

nitrile hydratase catalyzes the hydrolysis of the nitrile (or cyanohydrin) to the corresponding amide (or hydroxy acid)

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

amidase catalyzes the hydrolysis of the amide to the corresponding acid or hydroxy acid

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Implementation Method 3

some microorganisms have been shown to degrade aromatic nitriles by directly converting these nitriles to their respective acid through the action of nitrilase

Methodology Applied

Scientific EffectDecomposition (biological): Decomposition (biological)

Data Source

PatentUS7531343B2Induction and stabilization of enzymatic activity in microorganisms

Publication Date: 2009.05.12 CROWPIERCE TECH LLC

AI summary

The present invention is directed to methods for inducing desired activity in enzymes or microorganisms capable of producing the enzymes. The invention is further directed to methods of stabilizing activity in microorganisms. In specific embodiments, the invention provides methods for inducing and stabilizing nitrile hydratase activity, amidase activity, and asparaginase I activity. The invention further provides compositions comprising enzymes or microorganisms having induced and/or stabilized activity.