Modified Phenylalanine Dehydrogenase Mutations for Measurement Accuracy
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Solution Overview
Problem
Wild-type phenylalanine dehydrogenase has low substrate specificity and solubility, leading to inaccurate and segregated enzyme measurements, which are problematic for precise phenylalanine measurement in clinical diagnostics and food analysis.
Innovation Solution
A modified phenylalanine dehydrogenase with specific mutations in motifs such as GPALGGXRM, GRFXTGTDMGT, DF, GXANN, RH, and VNXGGLIQV, enhancing substrate specificity, solubility, and enzyme activity, is developed to improve phenylalanine measurement accuracy and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If wild-type phenylalanine dehydrogenase is used, then the enzyme can catalyze phenylalanine conversion, but substrate specificity is low and measurement accuracy deteriorates
Solution Approach 1:
The patent applies local quality by introducing specific amino acid mutations at particular positions in the enzyme's amino acid sequence (e.g., positions 41, 42, 43, 115, 124, 264, 271, 290, 294, 296, 297). These localized changes at specific motifs (GPALGGXRM, GRFXTGTDMGT, DF, GXANN, RH, VNXGGLIQV) selectively enhance substrate specificity for phenylalanine while maintaining catalytic activity, thereby resolving the contradiction between measurement accuracy and substrate specificity.
Solution Approach 2:
The patent employs parameter changes by modifying the amino acid sequence parameters of the enzyme through directed mutations. The mutations alter the enzyme's molecular properties to achieve higher substrate specificity and improved measurement accuracy, transforming the enzyme from a wild-type with low specificity to a modified version with optimized performance for phenylalanine measurement.
2Measurement precision
If wild-type phenylalanine dehydrogenase is used, then the enzyme maintains natural structure, but solubility is poor and enzyme segregation occurs
Solution Approach 1:
The patent applies local quality by introducing specific amino acid mutations at particular positions in the enzyme's amino acid sequence (e.g., positions 41, 42, 43, 115, 124, 264, 271, 290, 294, 296, 297). These localized changes at specific motifs (GPALGGXRM, GRFXTGTDMGT, DF, GXANN, RH, VNXGGLIQV) selectively enhance substrate specificity for phenylalanine while maintaining catalytic activity, thereby resolving the contradiction between measurement accuracy and substrate specificity.
Solution Approach 2:
The patent employs parameter changes by modifying the amino acid sequence parameters of the enzyme through directed mutations. The mutations alter the enzyme's molecular properties to achieve higher substrate specificity and improved measurement accuracy, transforming the enzyme from a wild-type with low specificity to a modified version with optimized performance for phenylalanine measurement.
3Productivity
If wild-type phenylalanine dehydrogenase is used, then the enzyme structure is maintained, but enzyme activity is insufficient for quick measurement
Solution Approach 1:
The patent applies local quality by introducing specific amino acid mutations at particular positions in the enzyme's amino acid sequence (e.g., positions 41, 42, 43, 115, 124, 264, 271, 290, 294, 296, 297). These localized changes at specific motifs (GPALGGXRM, GRFXTGTDMGT, DF, GXANN, RH, VNXGGLIQV) selectively enhance substrate specificity for phenylalanine while maintaining catalytic activity, thereby resolving the contradiction between measurement accuracy and substrate specificity.
Solution Approach 2:
The patent employs parameter changes by modifying the amino acid sequence parameters of the enzyme through directed mutations. The mutations alter the enzyme's molecular properties to achieve higher substrate specificity and improved measurement accuracy, transforming the enzyme from a wild-type with low specificity to a modified version with optimized performance for phenylalanine measurement.
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 provides improved substrate specificity, solubility, and activity, enabling quick, high-precision, and sensitive phenylalanine measurement, suitable for diagnosing phenylketonuria and analyzing phenylalanine content in food.
Implementation Method 1
an enzymatic method of measurement using a phenylalanine dehydrogenase derived from Thermoactinomyces intermedius
Data Source
AI summary
The present invention provides a unit and a method useful for more precise phenylalanine measurement. More specifically, the present invention provides a modified phenylalanine dehydrogenase that includes a mutation of at least one amino acid residue so as to improve the characteristics (for example, substrate specificity, solubility, and phenylalanine dehydrogenase activity) of a phenylalanine dehydrogenase related to measurement of phenylalanine, a method for analyzing phenylalanine by measuring phenylalanine contained in a test sample using the modified phenylalanine dehydrogenase, and others.