Trichoderma Protein Production with Low-Lactose Fed-Batch Induction

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Solution Overview

Problem

Existing methods for producing cellulolytic enzymes using Trichoderma reesei strains are inefficient due to high lactose costs, necessitating a reduction in lactose usage while maintaining protein production efficiency.

Innovation Solution

A method utilizing a Trichoderma reesei strain with a knocked-out cel1a gene, employing a composition with reduced lactose content (0-30% by weight) and additional sugars like glucose or xylose during the fed-batch phase to induce protein production.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If lactose is used as the sole inducing sugar in fed-batch fermentation, then protein production efficiency is maintained, but production costs increase due to high lactose price

Engineering Contradiction:

Improveprotein production efficiencyVSAvoidlactose usage amount

Core Design Contradiction:

ProductivityVSQuantity of substance

Solution Approach 1:

The invention changes the chemical composition parameters of the inducing sugar solution by replacing lactose with cheaper alternative sugars (glucose, xylose, maltose, sucrose) in specific proportions. This parameter substitution maintains the inducing effect on cellulase production while significantly reducing the cost per unit of protein produced

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The invention makes the sugar solution composition universally adaptable by providing a flexible formula that can accommodate multiple types of sugars (glucose, xylose, maltose, sucrose) in various proportions. This multi-functional sugar composition can be adjusted based on available raw materials and cost considerations while maintaining effective protein induction

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Quantity of substance

If alternative sugars (glucose, xylose) replace lactose in feed solution, then production costs decrease, but induction capacity is reduced

Engineering Contradiction:

Improvelactose contentVSAvoidinduction capacity

Core Design Contradiction:

Quantity of substanceVSProductivity

Solution Approach 1:

The invention creates a composite sugar solution containing multiple types of sugars (glucose, xylose, maltose, sucrose) in optimized proportions. This composite composition combines the cost advantages of alternative sugars with their synergistic inducing effects, achieving both cost reduction and maintained induction capacity that neither sugar alone could provide

Inventive Principle:

Principle #40Composite materials

Solution Approach 2:

The invention optimizes the concentration parameters of each sugar component in the feed solution. By adjusting the proportions of glucose, xylose, maltose, and sucrose within specific ranges, the inducing capacity is maximized while using cheaper alternative sugars instead of pure lactose

Inventive Principle:

Principle #35Parameter changes

3Productivity

If pure lactose is used for induction, then induction capacity is maximized, but production cost increases

Engineering Contradiction:

Improveinduction capacityVSAvoidproduction cost

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The invention replaces expensive lactose with cheaper alternative sugars that can be readily obtained from industrial byproducts (such as glucose from starch hydrolysis, xylose from hemicellulose). These cheaper sugars serve as effective substitutes for maintaining induction capacity while reducing raw material costs

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

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 method achieves equivalent protein production efficiency with significantly reduced lactose usage, dividing lactose amounts by 10 and maintaining specific productivity comparable to strains without the cel1a knockout.

Implementation Method 1

This fermentation method comprises two steps: a first step of fungus growth in the presence of an excess of carbon source and a second step of enzyme production due to the addition of an inducer into the medium with an optimized flow rate (fed-batch mode)

Methodology Applied

Scientific EffectFermentation: Fermentation

Implementation Method 2

a second step of producing proteins in a fed-batch phase in the presence of a composition comprising at least lactose and a second sugar, the lactose content in said composition representing approximately between 0 and 30% by weight of the total sugar content

Methodology Applied

Scientific EffectInduction:

Data Source

PatentUS12529045B2Method of producing proteins using a trichoderma fungus strain in which the CEL1A gene is invalidated

Publication Date: 2026.01.20 IFP ENERGIES NOUVELLES

AI summary

The invention relates to the various uses of a fungus strain which belongs to the Trichoderma genus and in which the cel1a gene has been knocked out. The invention relates in particular to a method of producing proteins by a fungus strain which belongs to the Trichoderma genus and in which the cel1a gene has been knocked out, comprising at least two steps:a first step of growth in a batch phase in the presence of at least one carbon growth substrate, anda second step of producing proteins in a fed-batch phase in the presence of a composition comprising at least lactose and a second sugar, the lactose content in the composition representing approximately between 0 and 30% by weight of the total sugar content of the composition, in particular between 5 and 30%.