Esterase Mutants for Thermally Stable Polyester Degradation
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Solution Overview
Problem
Existing esterases exhibit deficiencies in catalytic efficiency and thermal stability, preventing their industrial application for polyester degradation.
Innovation Solution
Development of esterase mutants with specific amino acid mutations at key sites, such as MT-1 to MT-42, which enhance polyester degradation activity and thermal stability, along with recombinant plasmids and strains for expression and purification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild-type esterase is used for polyester degradation, then polyester degradation activity is achieved, but catalytic efficiency and thermal stability are insufficient
Solution Approach 1:
The patent applies parameter changes by systematically mutating specific amino acid residues at positions 177, 178, 180, 181, and 208 in the esterase sequence. These parameter changes at the molecular level result in mutant esterases with improved thermal stability and catalytic efficiency, resolving the contradiction between reliability and productivity
Solution Approach 2:
The patent applies local quality by introducing specific amino acid substitutions at key positions in the esterase structure. Each mutation targets specific local regions of the enzyme to enhance particular properties (thermal stability or catalytic efficiency) while maintaining overall enzyme function, thereby resolving the contradiction between reliability and productivity
2Reliability
If wild-type esterase is used for polyester degradation, then polyester degradation activity is achieved, but thermal stability prevents industrial application
Solution Approach 1:
The patent applies parameter changes by systematically mutating specific amino acid residues at positions 177, 178, 180, 181, and 208 in the esterase sequence. These parameter changes at the molecular level result in mutant esterases with improved thermal stability and catalytic efficiency, resolving the contradiction between reliability and productivity
Solution Approach 2:
The patent applies local quality by introducing specific amino acid substitutions at key positions in the esterase structure. Each mutation targets specific local regions of the enzyme to enhance particular properties (thermal stability or catalytic efficiency) while maintaining overall enzyme function, thereby resolving the contradiction between reliability and productivity
3Stability of the object's composition
If esterase mutants with improved thermal stability are created, then thermal stability is enhanced, but catalytic efficiency must be maintained or improved
Solution Approach 1:
The patent applies parameter changes by systematically mutating specific amino acid residues at positions 177, 178, 180, 181, and 208 in the esterase sequence. These parameter changes at the molecular level result in mutant esterases with improved thermal stability and catalytic efficiency, resolving the contradiction between reliability and productivity
Solution Approach 2:
The patent applies local quality by introducing specific amino acid substitutions at key positions in the esterase structure. Each mutation targets specific local regions of the enzyme to enhance particular properties (thermal stability or catalytic efficiency) while maintaining overall enzyme function, thereby resolving the contradiction between reliability and 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 esterase mutants demonstrate improved thermal stability and catalytic efficiency, enabling polyester degradation across a wide pH range and elevated temperatures, specifically at 37°C to 70°C, with enhanced activity at pH 9.0 and 50°C compared to wild-type esterases.
Implementation Method 1
this esterase can hydrolyze polyesters
Implementation Method 2
Biological processes have emerged as a novel technology in recent years for the degradation and recycling of polyester plastics. It decomposes such organic matter under the action of biological entities (such as microorganisms, namely bacteria, fungi, and marine microalgae) or enzymes.
Data Source
AI summary
Disclosed in the present disclosure are esterase mutants having polyester degradation activity and the use thereof. The esterase mutants are one of the following: an esterase A as shown in SEQ ID NO. 3, of which the glutamic acid at the 177th site is mutated into glutamine; or the asparagine at the 178th site is mutated into alanine; or the serine at the 180th site is mutated into threonine, leucine, and valine; or the isoleucine at the 181st site is mutated into valine, etc. Experiments show that a protein expressed by the gene of the esterase mutant of the present disclosure can be correctly folded and can be purified in a large quantity in an Escherichia coli system.