Trichoderma Reesei Xylanase Production Without Enzyme Purification
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Solution Overview
Problem
Existing methods for producing xylooligosaccharides are complex, unsuitable for large-scale industrial production, and enzymes produced by certain microorganisms like Penicillium occitanis and Aspergillus niger are accompanied by toxin production, while Trichoderma strains yield enzyme mixtures that cannot be directly applied to xylooligosaccharide production.
Innovation Solution
A novel strain of Trichoderma reesei (BLCY-007) with high xylanase activity is developed, and a culture method involving simple centrifugation/filtration is used to obtain a crude enzyme preparation, which is then applied to produce xylooligosaccharides through enzymolysis, utilizing a combination of high-temperature and high-pressure treatment with xylanase from Trichoderma reesei to enhance extraction efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If xylanase is extracted and purified through salting out, dialysis, and column chromatography, then high purity xylanase is obtained, but the process becomes too complex for large-scale industrial production
Solution Approach 1:
The invention extracts and utilizes the xylanase activity directly from the fermentation broth of Trichoderma reesei without requiring complex purification steps. The crude enzyme preparation is applied directly to xylooligosaccharide production, taking out only the essential enzymatic function while leaving behind the complex purification process.
Solution Approach 2:
The Trichoderma reesei strain produces xylanase with high specific activity that functions effectively in the xylooligosaccharide production process without needing external purification. The enzyme preparation serves itself by being directly applicable from fermentation to application, eliminating the need for complex purification infrastructure.
2Productivity
If Penicillium occitanis or Aspergillus niger are used to produce xylanase, then significant yield is achieved, but toxins are co-produced creating safety hazards
Solution Approach 1:
The invention selects a microorganism that naturally produces only beneficial enzymes without harmful toxins. Trichoderma reesei converts the potential harm of toxin co-production into a benefit by being inherently toxin-free while maintaining high xylanase productivity, making the process safe for food and feed applications.
Solution Approach 2:
The invention applies the principle of local quality by selecting a specific strain (Trichoderma reesei) that has the localized property of producing high xylanase activity without the harmful property of toxin production. This strain-specific selection creates a locally optimized solution where only the desired enzymatic function is present.
3Ease of manufacture
If Trichoderma strains are used to produce xylanase, then the process is simplified, but enzyme mixtures are produced that cannot be directly applied to xylooligosaccharide production
Solution Approach 1:
The invention changes the parameter of enzyme specificity by selecting a Trichoderma reesei strain that naturally produces xylanase with high specificity for xylooligosaccharide production. This strain parameter change ensures that the simplified fermentation process yields an enzyme preparation with the precise specificity needed for direct application.
Solution Approach 2:
The invention extracts the beneficial property of high xylanase specificity from Trichoderma reesei while eliminating the harmful property of producing non-specific enzyme mixtures. The selected strain naturally produces a targeted enzyme profile that is directly applicable to xylooligosaccharide production without requiring separation from other enzymes.
4Productivity
If traditional methods are used to produce xylooligosaccharides, then production can proceed, but high production costs and environmental pollution result from extensive use of acids and alkalis
Solution Approach 1:
The invention replaces the mechanical/chemical system of acid and alkali treatment with a biological system using xylanase enzyme from Trichoderma reesei. This substitution eliminates the need for harsh chemicals while maintaining effective xylooligosaccharide production, thereby reducing environmental pollution and production costs.
Solution Approach 2:
The invention changes the chemical parameter approach from using strong acids and alkalis to using a biological catalyst (xylanase enzyme) that operates under milder conditions. This parameter change from chemical to biological catalysis reduces the harmful environmental factors associated with traditional methods while maintaining productivity.
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 high xylanase activity of 508 U/ml, simplifies the production process, reduces environmental impact, and enhances the extraction efficiency of xylooligosaccharides by minimizing the use of acids and alkalis, thereby lowering production costs.
Implementation Method 1
A strain of Trichoderma reesei BLCY-007, which has an Accession number: CGMCC No. 17970... The crude enzyme preparation obtained based on the Trichoderma reesei has a high xylanase activity... xylanase enzyme activity of 508 U/ml
Implementation Method 2
Xylooligosaccharides are a kind of oligosaccharide made from natural dietary fibers such as corncobs, cottonseed hulls, bagasse and other natural food fibers by saccharification and decomposition of hemicellulose with xylanase
Implementation Method 3
The production method thereof includes the following steps: a) culturing the cells... c) obtaining a culture supernatant from the culture... simple centrifugation/filtration is used to obtain a crude enzyme preparation
Data Source
AI summary
The invention relates to a strain of Trichoderma reesei BLCY-007 and its application in the production of xylooligosaccharides.