Trichoderma BXL1 Gene Disruption for Oxygen Saturation in Protein Culture

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

The challenge in scaling up protein production using filamentous fungi like Trichoderma is the increased energy and oxygen demand due to high viscosity of the culture medium, leading to decreased dissolved oxygen saturation and reduced productivity, especially when cellulose and xylan are used as inducers.

Innovation Solution

Disrupting the BXL1 gene in Trichoderma fungi to reduce oxygen uptake rates, allowing for higher concentrations of cellulose and xylan as inducers, which suppresses the decrease in dissolved oxygen saturation and enhances protein production.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If the size of the culture tank is increased for scaling up protein production, then the productivity is improved, but the oxygen-transfer coefficient decreases making it difficult to maintain dissolved oxygen saturation

Engineering Contradiction:

Improveprotein production volumeVSAvoiddissolved oxygen saturation

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The invention changes the genetic parameters of the fungus by disrupting the BXL1 gene encoding β-xylosidase. This genetic modification alters the metabolic characteristics of the fungus, reducing its oxygen uptake rate during cellulose and xylan degradation, thereby maintaining dissolved oxygen saturation in large-scale culture tanks without increasing stirring or oxygen supply

Inventive Principle:

Principle #35Parameter changes

2Reliability

If the number of stirring times is increased to maintain dissolved oxygen saturation in large tanks, then the oxygen supply is improved, but shear damage to fungal cells increases

Engineering Contradiction:

Improvedissolved oxygen saturationVSAvoidshear damage to fungal cells

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention changes the metabolic parameter of the fungus by disrupting the BXL1 gene, reducing oxygen uptake rate. This allows maintenance of dissolved oxygen saturation with reduced stirring frequency, thereby minimizing shear damage to fungal hyphae and cells

Inventive Principle:

Principle #35Parameter changes

3Reliability

If the amount of oxygen supply is increased to maintain dissolved oxygen saturation, then the oxygen availability is improved, but energy input increases

Engineering Contradiction:

Improvedissolved oxygen saturationVSAvoidenergy input for oxygen supply

Core Design Contradiction:

ReliabilityVSUse of energy by stationary object

Solution Approach 1:

The invention changes the metabolic parameter of the fungus by disrupting the BXL1 gene, reducing oxygen uptake rate during inducer degradation. This reduces the overall oxygen demand in the system, allowing maintenance of dissolved oxygen saturation with lower oxygen supply rates and reduced energy input

Inventive Principle:

Principle #35Parameter changes

4Productivity

If cellulose and xylan are added as inducers to increase protein production, then the productivity is improved, but the oxygen uptake rate increases causing further decrease in dissolved oxygen saturation

Engineering Contradiction:

Improveprotein productionVSAvoiddissolved oxygen saturation

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The invention extracts or removes the harmful function of β-xylosidase by disrupting the BXL1 gene. This eliminates the enzyme's ability to degrade xylan and consume oxygen, thereby allowing protein induction with cellulose and xylan without the detrimental increase in oxygen uptake rate

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention converts the harmful effect of xylan degradation (high oxygen consumption) into a beneficial outcome. By disrupting BXL1, the fungus can still utilize xylan-containing inducers for protein induction but without the oxygen-consuming degradation pathway, turning a harmful metabolic process into a beneficial one for maintaining dissolved oxygen saturation

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

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 results in increased protein concentration and productivity with improved dissolved oxygen saturation, facilitating efficient protein production and saccharification of biomass into xylo-oligosaccharides and glucose.

Implementation Method 1

using a fungus belonging to the genus Trichoderma in which the BXL1 gene encoding β-xylosidase has been disrupted decreases the oxygen uptake rate of the fungus belonging to the genus Trichoderma

Methodology Applied

Scientific EffectOxygen uptake: Aerobic Digestion

Data Source

PatentUS11453899B2Method of producing protein

Publication Date: 2022.09.27 TORAY INDUSTRIES INC

AI summary

A method produces a protein using a filamentous fungus, in which decrease in dissolved oxygen saturation during culture of the filamentous fungus can be suppressed even when the culture is scaled up. The method of producing a protein includes culturing a fungus belonging to the genus Trichoderma whose BXL1 gene was disrupted, using a biomass containing cellulose and xylan as an inducer. The use of the BXL1 gene-disrupted fungus belonging to the genus Trichoderma enables suppression of the decrease in dissolved oxygen saturation even when xylose and cellulose are used as inducers.