Protease Variants for Polyester Degradation
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Solution Overview
Problem
There is a need for proteases with improved activity and stability at high temperatures to enhance the efficiency of biodegradable plastic production and recycling processes, as existing proteases are not sufficient for efficient degradation of polyester materials like polylactic acid (PLA) at elevated temperatures.
Innovation Solution
Development of protease variants with specific substitutions such as X99F and X101L, along with additional substitutions like X3T, X97D, X103T, X104I, X127I, X161W, X161R, X161Y, X161L, X163R, X172K, X174G, X174V, X175A, X175Y, X175D, X175T, X175I, X176S, X194P, and X215K, which exhibit enhanced polyester degrading activity and stability, particularly for PLA.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If existing proteases are used for polyester degradation, then the process can proceed, but the activity and stability at high temperatures are insufficient for efficient degradation
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the protease through specific substitutions (X99F, X101L, and at least one additional substitution from the specified group). These molecular-level parameter changes result in improved thermal stability and polyester degradation efficiency, allowing the enzyme to maintain activity at elevated temperatures where existing proteases fail.
2Productivity
If protease variants with improved activity are developed, then polyester degradation efficiency increases, but the complexity of enzyme development and characterization increases
Solution Approach 1:
The patent applies local quality by making targeted, localized changes to specific amino acid positions (X99, X101, and one additional position from the specified group) rather than attempting global optimization. This focused approach to local sequence modification achieves improved degradation efficiency while limiting the overall complexity of the development process.
Solution Approach 2:
The invention systematically explores parameter changes at defined positions with specific amino acid substitutions. By pre-defining the positions and possible substitutions, the search space is constrained, making the development and characterization process more manageable while still achieving improved 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 protease variants demonstrate improved polyester degrading activity and stability, enabling more efficient degradation of PLA and other polyesters, thereby improving the competitiveness of biodegradable plastic production and recycling processes.
Implementation Method 1
Proteases are able to catalyze the hydrolysis of a variety of polymers, including polyesters. In this context, proteases have shown promising effects in several industrial applications... enzymes are able to accelerate hydrolysis of polyester containing material, and more particularly of plastic products, even down to the monomer level
Data Source
AI summary
The present invention relates to protease variants having polyester degrading activity, polynucleotides encoding said variants, nucleic acid constructs and expression vectors comprising said polynucleotides, host cells expressing said variants, compositions comprising the variants, methods of producing the variants, and use of the variants.