Peptide Tag Composition for Higher Expression With Minimal Structural Impact
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing peptide tag techniques do not effectively enhance the expression level of proteins of interest, particularly in host cells like E. coli and Brevibacillus, and their impact on protein structure and function is unclear.
Innovation Solution
Development of peptide tags with specific amino acid substitutions, such as replacing serine and glycine with basic or acidic amino acids, to improve protein expression levels by linking them to proteins of interest, utilizing sequences like Xm(PYn)qPZr, where X, Y, and Z represent specific amino acids, and m, n, and r represent the number of amino acids, enhancing expression without significantly affecting protein structure.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing peptide tag techniques are used to link proteins, then solubility is improved and inclusion body formation is suppressed, but expression level is not enhanced
Solution Approach 1:
The invention changes the amino acid composition parameters of the peptide tag by incorporating specific sequences with defined proportions of proline, glycine, serine, and other amino acids. The peptide tag follows the sequence formula X1 to X50 where specific amino acids are positioned to optimize both solubility and expression level, resolving the contradiction between reliability and productivity
2Reliability
If peptide tags are linked to proteins of interest, then normal expression is promoted, but the impact on protein structure and function remains unclear
Solution Approach 1:
The peptide tag is designed with local quality optimization where specific amino acid residues are positioned at specific locations in the sequence. The tag contains functional elements like proline-rich regions for solubility while maintaining minimal interference with the protein of interest structure, allowing normal expression while preserving protein integrity
3Productivity
If toxin fusion proteins are expressed in plants using peptide linkers with prolines, then high-level accumulation is achieved, but the applicability to single protein expression systems is limited
Solution Approach 1:
The peptide tag is designed as a universal element that can be applied to various protein expression systems including single protein expression, toxin fusion proteins, and different host organisms. The standardized sequence format X1 to X50 with defined amino acid composition makes it adaptable across multiple applications while maintaining high-level accumulation capability
Data Source
AI summary
A peptide comprising the sequence shown below is added as a peptide tag to a useful protein, followed by allowing its expression.Xm(PYn)qPZr In this formula, X, Y, and Z each represent an amino acid residue independently selected from the group consisting of R, G, S, K, T, L, N, Q, and H, with the proviso that at least one Y represents K, L, N, Q, H, or R. m represents an integer of 0 to 5; n represents 1, 2, or 3; q represents an integer of 1 to 10; and r represents an integer of 0 to 10.