Mutant L-amino Acid Alpha-ligase for Imidazole Dipeptide Synthesis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
There is a lack of a simple, easy, and low-cost method for producing imidazole dipeptides such as carnosine, anserine, and balenine, which have known antifatigue, antioxidant, blood pressure regulation, anti-inflammatory, and uric acid reduction effects, limiting their availability as supplements.
Innovation Solution
A mutant L-amino acid α-ligase enzyme with specific amino acid substitutions is used to enhance the production efficiency of imidazole dipeptides, specifically carnosine, anserine, and balenine, by forming α-peptide bonds, and a host cell with a nucleic acid encoding this protein is employed to facilitate their production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If conventional production methods are used, then imidazole dipeptides can be produced, but the production process is complex and costly
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the ligase enzyme through site-specific mutagenesis. Specific amino acid residues are substituted to enhance the enzyme's synthetic activity for imidazole dipeptides, transforming the production process from complex and low-yield to simple and high-yield through biochemical parameter optimization
2Ease of manufacture
If conventional production methods are used, then imidazole dipeptides can be produced, but the production cost is high
Solution Approach 1:
The patent changes the biochemical parameters of the production system by introducing mutant ligase enzymes with improved catalytic properties. These parameter changes in enzyme structure and function directly increase dipeptide yield while reducing production costs through more efficient biocatalysis
3Reliability
If wild-type ligase is used, then basic peptide synthesis occurs, but imidazole dipeptide synthetic activity is low
Solution Approach 1:
The patent applies local quality by making specific localized changes to the ligase enzyme structure through site-specific mutagenesis. Rather than changing the entire enzyme, only particular amino acid residues are substituted to enhance imidazole dipeptide synthesis while maintaining overall peptide synthesis capability
Solution Approach 2:
The patent changes specific biochemical parameters of the ligase enzyme through amino acid substitution. These parameter changes in enzyme structure directly improve catalytic activity for imidazole dipeptides while preserving the fundamental peptide synthesis function
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 mutant enzyme significantly improves the synthetic activity of imidazole dipeptides, enabling their production in high efficiency and low cost, particularly for carnosine, using a peptidase-deficient host cell, ensuring high yields and ease of production.
Implementation Method 1
L-amino acid α-ligase is an enzyme catalyzing peptide synthesis by forming α-peptide bond as a substrate
Implementation Method 2
catalyzing peptide synthesis by forming α-peptide bond
Data Source
AI summary
The present invention provides L-amino acid α-ligase represented by sequence number 1 (SEQ ID NO. 1), which is a mutant protein including substitution of at least 1-3 amino acid residues of amino acid sequence of protein YwfE, with the substitution being at least one of an asparagine (N) residue of the 108th place from the N terminal being substituted with at least one of an alanine (A) residue, a glutamic acid (E) residue and a glutamine (Q) residue, an isoleucine (I) residue of the 112th place from the N terminal being substituted with a valine (V) residue, and a histidine (H) residue of the 378th place from the N terminal being substituted with at least one of a lysine (K) residue or an arginine (R) residue.