Modified Glycine Oxidase Enzyme Stability and Specificity
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Solution Overview
Problem
Current methods for measuring glycine, such as those using glycine oxidase, face challenges with low activity and stability, and lack specificity for glycine, making practical measurement and kit development difficult.
Innovation Solution
A modified glycine oxidase is developed by mutating specific amino acid residues in the enzyme's sequence, improving its activity, thermal stability, and substrate specificity, allowing for enhanced glycine measurement and production of glyoxylic acid.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a glycine oxidase is used for glycine measurement, then the measurement can be performed using an enzyme-based method, but the enzyme exhibits low activity and low stability
Solution Approach 1:
The patent applies parameter changes by mutating specific amino acid residues in the glycine oxidase sequence to alter the enzyme's physical and chemical properties. The mutations are designed to improve both stability and activity parameters simultaneously, resolving the contradiction between these two properties that were previously inversely related in natural glycine oxidases.
Solution Approach 2:
The patent applies local quality by introducing specific mutations at particular positions (TTS motif, HCY motif, LRP motif, GML motif, SG motif, PGT motif) in the enzyme sequence. Each motif represents a local region with specific functional properties, and targeted mutations in these regions selectively improve enzyme performance without compromising overall structure.
2Measurement precision
If a glycine oxidase is used for glycine measurement, then the measurement can be performed enzymatically, but the substrate specificity for glycine is insufficient
Solution Approach 1:
The patent applies local quality by introducing specific mutations at particular positions (TTS motif, HCY motif, LRP motif, GML motif, SG motif, PGT motif) in the enzyme sequence. Each motif represents a local region with specific functional properties, and targeted mutations in these regions selectively improve enzyme performance without compromising overall structure.
Solution Approach 2:
The patent applies parameter changes by mutating specific amino acid residues in the glycine oxidase sequence to alter the enzyme's physical and chemical properties. The mutations are designed to improve both stability and activity parameters simultaneously, resolving the contradiction between these two properties that were previously inversely related in natural glycine oxidases.
3Measurement precision
If conventional amino acid analysis methods are used, then accurate measurement can be achieved, but the instruments are large-sized, expensive, and require technical knowledge for operation and maintenance
Solution Approach 1:
The patent applies mechanics substitution by replacing complex mechanical instrumentation (amino acid analyzers, HPLC systems, LC-MS) with a biochemical system based on modified glycine oxidase. This enzyme-based approach eliminates the need for sophisticated mechanical equipment while maintaining measurement capability, thereby reducing device complexity and operational requirements.
Solution Approach 2:
The patent applies copying by creating a simplified biochemical replica of the measurement function. Instead of using complex instruments to detect and measure amino acids, the modified enzyme copies the analytical function through its catalytic activity, which can be detected by simple means such as colorimetric or fluorometric assays.
4Productivity
If conventional amino acid analysis methods are used, then measurement can be performed, but it takes a long time to analyze many specimens due to sequential analysis requirement
Solution Approach 1:
The patent applies mechanics substitution by replacing complex mechanical instrumentation (amino acid analyzers, HPLC systems, LC-MS) with a biochemical system based on modified glycine oxidase. This enzyme-based approach eliminates the need for sophisticated mechanical equipment while maintaining measurement capability, thereby reducing device complexity and operational requirements.
Solution Approach 2:
The patent applies preliminary action by preparing the modified glycine oxidase in advance with improved properties (stability, activity, specificity) so that it can be directly used for rapid measurement without requiring time-consuming instrument setup, calibration, or sequential processing. The enzyme is pre-optimized to function immediately under assay conditions.
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 enzyme enables rapid, sensitive, and specific measurement of glycine, with improved thermal stability, making it suitable for various applications including biological investigation, health nutrition, and food manufacture.
Implementation Method 1
A modified glycine oxidase is developed by mutating specific amino acid residues in the enzyme's sequence, improving its activity, thermal stability, and substrate specificity
Implementation Method 2
A glycine oxidase is known as an enzyme that acts upon glycine
Data Source
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AI summary
The present invention provides a means and method useful for measurement of a glycine concentration. Specifically, the present invention provides a modified enzyme in which at least one amino acid residue is mutated so as to improve a property of a glycine oxidase which is associated with the measurement of glycine (e.g., activity of glycine oxidase for glycine, thermal stability of glycine oxidase, and substrate specificity of glycine oxidase for glycine,); and a method of analyzing glycine, comprising measuring glycine contained in a test sample using the modified enzyme; and the like.