Filamentous Fungi Protein Production via Regulatory Gene Modification

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Solution Overview

Problem

Current methods for enhancing the production of cellulases and hemicellulases in filamentous fungi like Trichoderma lack efficiency, as existing regulatory mechanisms do not adequately support increased production of desired enzymes like CBHI, EGI, or xylanase.

Innovation Solution

Genetic modification of Trichoderma hosts to overexpress specific genes, such as tre122523, or modify regulatory genes to enhance the production of CBHI, EGI, or xylanase by altering the expression levels of these enzymes, thereby increasing enzymatic activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional regulatory mechanisms are used for enzyme production in Trichoderma, then production process is maintained with standard biomass consumption, but enzymatic activity and production efficiency remain insufficient

Engineering Contradiction:
Improveenzymatic activityVSAvoidbiomass consumption
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The patent applies parameter changes by modifying the expression levels of regulatory genes (such as xlnR, celR, and other transcription factors) to alter the production parameters of target enzymes. By changing the regulatory gene expression parameters, the system achieves increased enzymatic activity (CBHI, EGI, xylanase) while optimizing biomass consumption, thus resolving the contradiction between productivity and quantity of substance.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses regulatory genes as intermediaries to mediate between the cultivation conditions and the target enzyme production. By introducing or modifying regulatory genes (xlnR, celR, etc.), these intermediaries control and enhance the expression of cellulase and hemicellulase genes, enabling increased enzymatic activity without proportional increases in biomass consumption.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Productivity

If genetic modification is applied to overexpress regulatory genes, then enzymatic activity increases, but process complexity increases

Engineering Contradiction:
Improveprotein production efficiencyVSAvoidgenetic modification complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent extracts and utilizes specific regulatory genes (such as xlnR, celR) that control enzyme production pathways. By isolating and overexpressing these key regulatory genes, the invention simplifies the approach to enhancing protein production efficiency, avoiding the need for complex modifications of entire metabolic pathways while achieving the desired productivity increase.

Inventive Principle:
Principle #2Taking out (Extraction)

3Speed

If standard cultivation conditions are used, then nutrient consumption is high, but production speed is slow

Engineering Contradiction:
Improveproduction speedVSAvoidnutrient consumption
Core Design Contradiction:
SpeedVSUse of energy by moving object

Solution Approach 1:

The patent changes the physiological parameters of the Trichoderma host by modifying regulatory gene expression, which in turn alters nutrient utilization patterns. The modified strains exhibit improved production speed with optimized nutrient consumption by changing the regulatory control parameters of enzyme synthesis, allowing faster production without proportional increases in nutrient use.

Inventive Principle:
Principle #35Parameter changes

Data Source

PatentEP2733210B1Method for improved protein production in filamentous fungi
Publication Date: 2017.12.06 AB ENZYMES OY
  • EP2733210B1 patent drawingFigure 1A~1B
  • EP2733210B1 patent drawingFigure 1C~1D
  • EP2733210B1 patent drawingFigure 1E~1F

AI summary

The present invention relates to a method for genetically modifying a filamentous fungus host for improved protein production. The method comprises that a filamentous fungus host is genetically modified to overexpress or to be deficient of specific genes. The invention relates also to the modified hosts. Furthermore, the invention relates to a method for improved production or for producing an improved composition of proteins, such as cellulases, hemicellulases, other proteins involved in the degradation of lignocellulosic material, or other proteins, in a filamentous fungus host.