GH7 Endoglucanase Thermostabilization via Amino Acid Mutations

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Solution Overview

Problem

Current endoglucanases from the GH7 family are not thermostable enough for industrial applications, particularly at elevated temperatures, which limits their effectiveness in processes like saccharification and fiber modification.

Innovation Solution

Development of endoglucanase proteins with amino acid sequences showing at least 96% identity to a specific sequence (SEQ. ID NO. 2) that maintain at least 90% residual substrate conversion activity at 60°C for one hour, exhibiting active thermostabilization and improved temperature stability.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Temperature

If endoglucanases from GH7 family are used for industrial applications, then substrate conversion activity is achieved, but thermal stability is insufficient at elevated temperatures

Engineering Contradiction:
Improvethermal stabilityVSAvoidenzyme activity maintenance
Core Design Contradiction:
TemperatureVSReliability

Solution Approach 1:

The patent applies parameter changes by modifying amino acid sequences of endoglucanases to alter their thermal stability parameters. Specific mutations in the protein structure change the enzyme's physical properties, enabling it to maintain activity at elevated temperatures while preserving catalytic function. This directly resolves the contradiction between thermal stability and enzyme activity maintenance.

Inventive Principle:
Principle #35Parameter changes

2Productivity

If temperature is increased for industrial processes, then processing efficiency is improved, but enzyme stability decreases

Engineering Contradiction:
Improveprocessing efficiencyVSAvoidenzyme stability
Core Design Contradiction:
ProductivityVSStability of the object's composition

Solution Approach 1:

The invention changes the biochemical parameters of the enzyme through amino acid mutations, allowing the enzyme to withstand higher processing temperatures. This enables industrial processes to run at elevated temperatures for improved productivity while the modified enzyme maintains its structural stability and catalytic activity under these conditions.

Inventive Principle:
Principle #35Parameter changes

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 resulting endoglucanases demonstrate enhanced thermal stability and active thermostabilization, allowing them to maintain high substrate conversion activity at elevated temperatures, thus improving their performance in industrial processes such as lignocellulose saccharification and fiber modification.

Implementation Method 1

Cellulases comprise a structurally and functionally diverse class of glycohydrolases acting on cellulose. Having in common the hydrolytic cleavage activity of glycosidic bonds present in cellulose polymers or oligomers

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentEP2660319B1Endoglucanases with improved properties
Publication Date: 2016.09.14 CLARIANT PRODUKTE (DEUTSCHLAND) GMBH GROUP INTELLECTUAL PROPERTY
  • EP2660319B1 patent drawingFigure 1~2
  • EP2660319B1 patent drawingFigure 3~4
  • EP2660319B1 patent drawingFigure 5~6

AI summary

The present invention relates to thermostable endoglucanases, particularly to proteins having endoglucanase activity which comprises an amino acid sequence having at least 96% identity to SEQ. ID NO.: 2, and proteins having endoglucanase activity which belongs to the GH7 class and which shows active thermostabilization.