Endoglucanase PPCE Expression for Acidic Low-Temperature Fabric Cellulase
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Solution Overview
Problem
Current cellulase preparations for fabric processing lack enzymes with low optimum temperature and strongly acidic pH, which are desired for industrial applications, and face challenges in overexpression and stability during production.
Innovation Solution
A novel endoglucanase, PPCE, derived from Penicillium pinophilum PF1365, with a low optimum temperature and pH, is isolated and produced in Trichoderma viride using a regulatory sequence from the cbhI gene, creating a cellulase preparation with high activity and improved stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If endoglucanase from genus Trichoderma is used for fabric processing under acidic conditions, then synergistic effect and productivity are improved, but gene expression is inhibited due to codon usage differences
Solution Approach 1:
The patent modifies the nucleotide sequence of the exogenous endoglucanase gene to match the codon usage preferences of the Trichoderma host organism. This codon optimization allows the foreign gene to be properly translated by the host's cellular machinery, resolving the expression inhibition problem while maintaining the desired acidic cellulase activity and high productivity
2Productivity
If endoglucanase with high activity is used for fabric processing, then fabric treatment effect is improved, but enzyme digestion by proteases during cultivation increases
Solution Approach 1:
The patent employs a secretion signal peptide as an intermediary element that directs the synthesized endoglucanase to be secreted into the culture medium. This spatial separation protects the high-activity enzyme from intracellular proteases during cultivation, maintaining enzyme stability and integrity while preserving its high catalytic activity for fabric processing applications
3Adaptability or versatility
If conventional endoglucanases are used for fabric processing, then fabric treatment is achieved, but low optimum temperature and strongly acidic pH enzymes are unavailable
Solution Approach 1:
The patent successfully isolates and characterizes endoglucanase variants with shifted operational parameters - specifically those exhibiting low optimum temperatures and strongly acidic pH optima. By modifying the enzyme's amino acid sequence through mutagenesis or selecting natural variants, the patent creates enzymes adapted to operate efficiently under conditions previously unavailable, thereby expanding industrial applicability while maintaining reliable fabric treatment performance
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
PPCE exhibits high activity in fabric processing, improving fabric appearance and texture, and is produced at a lower cost, addressing the industrial need for low-temperature and strongly acidic enzymes, with enhanced stability and efficiency.
Implementation Method 1
Endoglucanase PPCE, having at the N-terminus thereof the amino acid sequence of SEQ ID NO: 2 or an amino acid sequence in which one amino acid is deleted, substituted, or added in the amino acid sequence of SEQ ID NO: 2
Data Source
AI summary
A novel endoglucanase PPCE derived from Penicillium pinophilum, a cellulase preparation containing the endoglucanase PPCE, and a method of treating a cellulose-containing fabric utilizing the endoglucanase PPCE or the cellulase preparation, are disclosed. The endoglucanase PPCE is highly active to a fabric, and has a low optimum temperature and a strongly acidic optimum pH.