Esterase OXY+1 Residue Modification for Perhydrolase Activity
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Solution Overview
Problem
Current enzymes, such as esterases and lipases, lack sufficient perhydrolase activity, limiting their ability to catalyze reversible peracid formation from hydrogen peroxide and carboxylic acids, and there is a need to enhance or introduce this activity for industrial applications like laundry detergents and waste treatment.
Innovation Solution
Modifying the nucleic acid encoding esterases or lipases by altering the codon encoding the OXY+1 residue to encode amino acids that can adopt a cis-configuration, such as proline, to increase or introduce perhydrolase activity, thereby shifting their catalytic activity from hydrolysis to perhydrolysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If esterase or lipase enzymes are used in their natural form, then they exhibit sufficient esterase and/or lipase activity, but they lack sufficient perhydrolase activity
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence at the OXY+1 position of the enzyme. Specifically, it changes the residue to one capable of cis-configuration (proline, hydroxyproline, or other cis-forming residues), which alters the local structural parameters of the active site. This structural parameter change enables the enzyme to accommodate and catalyze perhydrolysis reactions, thereby improving perhydrolase activity and catalytic efficiency for peracid formation.
2Reliability
If the enzyme activity is shifted from hydrolysis to perhydrolysis, then perhydrolase activity increases, but esterase activity may be reduced
Solution Approach 1:
The patent applies local quality by making a targeted modification at a specific location in the enzyme - the OXY+1 residue in the active site. This localized change affects the chemical properties (introducing cis-configuration capability) at that specific position, which then influences the overall catalytic behavior. The modification is confined to a single residue position, allowing the rest of the enzyme structure to maintain its original properties while gaining new catalytic capability.
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 modified enzymes exhibit a significant increase in perhydrolase activity, up to 28-fold, while maintaining or reducing esterase activity, allowing for efficient catalysis of peracid formation and potential use in industrial processes like laundry detergents and waste treatment.
Implementation Method 1
catalyze reversible peracid formation from hydrogen peroxide and carboxylic acids
Data Source
AI summary
The invention provides for methods of modifying n esterase or a lipase enzyme to introduce perhydrolase activity or to increase the already-existing perhydrolase activity. The invention also provides for methods of converting an polypeptide having predominantly esterase and/or lipase catalytic activity to a polypeptide having predominantly perhydrolase activity.