NCgl0581E136P Mutant for Low-Concentration L-Serine Detection

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Solution Overview

Problem

Wild-type L-serine biosensors based on transcription factor NCgl0581 have a high detection limit, making them unsuitable for analyzing low-concentration L-serine.

Innovation Solution

A mutant form of NCgl0581, designated NCgl0581E136P, is created by mutating the 136th glutamic acid to proline, reducing the detection limit and enhancing sensitivity through molecular docking and virtual screening.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If wild-type transcription factor NCgl0581 is used to construct L-serine biosensor, then the biosensor can detect L-serine, but the detection limit is high and sensitivity is low

Engineering Contradiction:

Improvedetection limitVSAvoidsensitivity

Core Design Contradiction:

Measurement precisionVSReliability

Solution Approach 1:

The patent applies parameter changes by mutating the amino acid sequence of the transcription factor NCgl0581 at position 136 (E136P mutation). This genetic parameter modification alters the protein's binding characteristics to L-serine, resulting in a biosensor with reduced detection limit (from 50 mM to 25 mM) and enhanced sensitivity, directly resolving the technical contradiction between detection limit and sensitivity

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent implements local quality by introducing a specific point mutation (E136P) at a critical binding site of the transcription factor. This localized modification at position 136 of the amino acid sequence selectively enhances the protein's affinity for L-serine without compromising overall protein structure or function, thereby improving detection performance while maintaining biosensor operational reliability

Inventive Principle:

Principle #3Local quality

2Reliability

If traditional static improvement methods (adjusting promoter strength, RBS) are used, then biosensor performance can be improved, but the methods have limited data support and poor generality

Engineering Contradiction:

Improvebiosensor performanceVSAvoidgenerality

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent transitions from traditional static parameter adjustment (promoter strength, RBS) to rational design-based parameter optimization through in silico mutagenesis. By using computational tools to predict and evaluate mutation effects on transcription factor-L-serine binding, the method achieves improved biosensor performance with broader applicability and stronger theoretical foundation, resolving the contradiction between performance improvement and method generality

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS12448422B2Transcription factor NCgl0581 mutant and use thereof in L-serine detection

Publication Date: 2025.10.21 YIXING INST OF FOOD & BIOTECHNOLOGY CO LTD

AI summary

The present invention provides a transcription factor NCgl0581 mutant and use thereof in L-serine detection, belonging to the technical field of biological detection. According to the present invention, key binding sites of the transcription factor NCgl0581 with L-serine are obtained through molecular docking, and further model analysis and virtual screening are carried out to obtain an L-serine biosensor mutant NCgl0581E136P. Experiments show that the biosensor constructed based on the mutant NCgl0581E136P can respond to 25 mM serine, which is half of the response concentration before the mutation. The present invention reduces the detection limit of the biosensor through mutation, lays a foundation for the detection of low-concentration L-serine, and has good application value and prospect.