Enzyme Production Substrate Mixture for Trichoderma reesei

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Solution Overview

Problem

Current methods for producing cellulolytic and hemicellulolytic enzymes are costly due to high lactose prices and low productivity with existing substrates, necessitating the development of novel inducing substrates that perform as well as lactose at a lower cost.

Innovation Solution

A process using a mixture of glucose, lactose, and xylose as the inducing substrate, with specific weight ratios, to enhance enzyme production in Trichoderma reesei strains, allowing for higher protein concentrations and improved xylanase activity, while reducing costs by utilizing co-products from lignocellulosic biomass processing.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If lactose is used as the inducing substrate for cellulase production, then enzyme productivity is improved, but production cost increases due to high lactose prices

Engineering Contradiction:
Improveenzyme productivityVSAvoidproduction cost
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent combines multiple carbon sources (glucose, xylose, and lactose) into a mixed substrate formulation. This merging approach allows the system to benefit from the high inducement efficacy of lactose while diluting its cost impact through inclusion of cheaper alternatives like glucose and xylose, thereby resolving the contradiction between productivity and production cost

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The invention uses a composite substrate composed of multiple sugar types (glucose, xylose, and lactose) in specific ratios. This composite approach leverages the synergistic effects of different carbon sources to maintain high cellulase induction while reducing dependence on expensive lactose, thus achieving both high productivity and cost-effectiveness

Inventive Principle:
Principle #40Composite materials

2Productivity

If existing substrates are used for enzyme production, then process simplicity is maintained, but productivity remains low

Engineering Contradiction:
Improveenzyme productivityVSAvoidsubstrate composition complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent optimizes the compositional parameters of the substrate by specifying precise ratios of glucose, xylose, and lactose. This parameter optimization allows the system to achieve enhanced productivity through controlled substrate composition while maintaining manageable process complexity through defined formulation specifications

Inventive Principle:
Principle #35Parameter changes

3Speed

If glucose is used as the carbon source, then rapid growth of Trichoderma reesei is achieved, but catabolic repression reduces cellulase production

Engineering Contradiction:
Improvegrowth rateVSAvoidcellulase production
Core Design Contradiction:
SpeedVSProductivity

Solution Approach 1:

The patent employs a dynamic substrate composition that evolves during the fermentation process. The mixed sugar formulation allows the system to exploit glucose for rapid initial growth while the presence of lactose and xylose provides continuous induction signals for cellulase production, thereby dynamically balancing growth rate and enzyme productivity throughout the process

Inventive Principle:
Principle #15Dynamics

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 process achieves protein concentrations 50-60% higher than with lactose alone and 3 times higher than with improved xylose-rich mixtures, significantly increasing cellulase and xylanase activities, thus reducing production costs and optimizing enzyme productivity.

Implementation Method 1

Glucose exerts a catabolic repression effect on the production of cellulases

Methodology Applied
Scientific EffectCatabolic repression:

Implementation Method 2

In the presence of an inducing substrate, wild type strains of Trichoderma reesei have the ability to secrete an enzymatic complex which is well suited to cellulose hydrolysis

Methodology Applied
Scientific EffectEnzyme induction:

Implementation Method 3

The enzymatic hydrolysis step can be used to transform cellulose and hemicelluloses into sugars using cellulolytic and/or hemicellulolytic enzymes

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 4

Glucose may, for example, be readily transformed into ethanol by the yeast Saccharomyces cerevisiae during the alcoholic fermentation step

Methodology Applied
Scientific EffectFermentation: Fermentation

Data Source

PatentUS10457925B2Process for the production of cellulolytic and/or hemicellulolytic enzymes
Publication Date: 2019.10.29 IFP ENERGIES NOUVELLES

AI summary

The process for the production of cellulolytic and/or hemicellulolytic enzymes by a cellulolytic and/or hemicellulolytic microorganism according to the present invention comprises at least one phase for growth in the presence of a source of carbon and at least one phase for production in the presence of an inducing substrate, in which said inducing substrate is a mixture comprising 40% to 65% by weight of glucose or cellulosic hydrolysates, 21% to 25% by weight of lactose and 10% to 39% by weight of xylose or a solution of a lignocellulosic hemicellulosic hydrolysate, the sum of these three constituents being equal to 100%.