Mutant Trichoderma reesei Strain for High Cellulase Production
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Solution Overview
Problem
Current methods for producing cellulase enzymes, particularly from Trichoderma strains, face challenges in achieving high enzyme activity and stability, with existing strains often requiring optimal conditions that can be difficult to maintain and resulting in variable fermentation yields.
Innovation Solution
A mutant strain of Trichoderma reesei, designated as Trichoderma reesei 601-17, is developed, which exhibits high cellulase activity at pH 3.0-6.0 and temperatures between 23-35°C, and a method involving seed fermentation followed by transfer to a larger fermentation medium at controlled temperatures and pH for extended periods to produce cellulase with specific enzyme activities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional Trichoderma strains are used for cellulase production, then fermentation can be performed with standard procedures, but the enzyme activity and production yield remain variable and suboptimal
Solution Approach 1:
The patent applies parameter changes by optimizing fermentation conditions including temperature (28-35°C), pH (4.5-6.5), and extended fermentation time (120-168 hours) to achieve high and stable cellulase production. The mutant strain Trichoderma reesei 601-17 was developed with improved enzyme activity under these optimized parameters, resolving the contradiction between productivity and reliability.
2Productivity
If optimal conditions are maintained for high cellulase activity, then enzyme production is maximized, but the complexity of maintaining optimal conditions increases
Solution Approach 1:
The mutant strain Trichoderma reesei 601-17 exhibits self-service characteristics by maintaining high cellulase production capability under a broad range of conditions (pH 3.0-6.0, temperature 23-35°C). The strain autonomously adapts to varying fermentation conditions without requiring precise control, thereby achieving high productivity while reducing operational complexity.
3Quantity of substance
If extended fermentation time is used to maximize cellulase production, then enzyme yield increases, but the fermentation process duration and resource consumption increase
Solution Approach 1:
The patent employs preliminary action through a two-stage fermentation process: seed fermentation (72-96 hours) followed by production fermentation (48-72 hours). This staged approach prepares the culture in advance with optimized growth conditions, enabling the main fermentation to achieve high cellulase yields in reduced time, thus resolving the time-yield contradiction.
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 mutant strain Trichoderma reesei 601-17 achieves high enzyme activities of Filter Paper Activity (792 U/mL), Endo-1,4-β-D-glucanase (1389 U/mL), and β-1,4-glucosidase (486 U/mL), maintaining genetic stability across generations and optimizing fermentation conditions for enhanced cellulase production.
Implementation Method 1
The cellulase is applied in a food industry and an environmental industry extensively. Trichoderma reesei (the asexual anamorph of Hypocrea jecorina) is an important industrial source of cellulase and hemicellulase enzymes. The term cellulase (or cellulase enzymes) broadly refers to enzymes that catalyze the hydrolysis of beta-1,4-glucosidic bonds joining individual glucose units in cellulose polymers.
Implementation Method 2
The cellulase catalyzes the hydrolysis of beta-1,4-glucosidic bonds joining individual glucose units in cellulose polymers. The EG acts on insoluble cellulose surfaces, breaks internal bonds to disrupt crystalline structures of the cellulose and expose individual cellulose polysaccharide chains.
Implementation Method 3
The cellulase used for industrial production is prepared from fungi, typically Trichoderma (Trichodema), Aspergillus (Aspergillus), and Penicillium (Penicillium) fermentation.
Data Source
AI summary
The present invention relates to a mutant strain of Trichodema reesei, namely, CCTCC No: M 2013540, that produces cellulase with high enzyme activity, and a method of producing thereof. The enzyme activity of the cellulase was as follows: Filter Paper Activity (FPA): 792 U/mL, Endo-1,4-β-D-glucanase (EG): 1389 U/mL, Exo-1,4-β-D-glucannase (CBH): 680 U/mL, β-1,4-glucosidase (BG): 486 U/mL.