Recombinant Polypeptide for Staphylococcal Cell Wall Lysis
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Solution Overview
Problem
Current methods for treating staphylococcal infections, particularly those caused by antibiotic-resistant Staphylococcus aureus, are inadequate due to side effects and the inability of existing bacteriolytic enzymes to maintain activity in physiological conditions, especially in raw milk and serum, limiting their effectiveness in treating mastitis and other dermatological disorders.
Innovation Solution
A recombinant polypeptide combining the LytM catalytic domain with the lysostaphin cell wall binding domain, optionally linked by a peptide linker, which exhibits strong antibacterial activity against Staphylococcus strains under physiological conditions, effectively binding to and lysing bacterial cell walls without requiring bacterial growth or metabolic activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing bacteriolytic enzymes are used to treat staphylococcal infections, then antibacterial activity is achieved, but the enzymes fail to maintain activity in physiological conditions such as raw milk and serum
Solution Approach 1:
The invention creates a chimeric enzyme by combining the catalytic domain of LytM (residues 185-316) with the cell wall binding domain of lysostaphin through genetic fusion. This composite structure integrates the bacteriolytic activity of LytM with the target-specific binding capability of lysostaphin, enabling the enzyme to maintain stability and activity in physiological conditions like raw milk and serum while effectively lysing staphylococcal cell walls.
2Reliability
If antibiotics are used to treat staphylococcal infections, then bacterial growth is inhibited, but side effects and antibiotic resistance emerge
Solution Approach 1:
The invention replaces the chemical mechanism of antibiotics (which inhibit bacterial metabolism and gene expression) with a mechanical/enzymatic mechanism that directly cleaves peptidoglycan cross-links in the bacterial cell wall. This bacteriolytic approach physically disrupts the cell wall structure, providing a fundamentally different mode of action that bypasses antibiotic resistance mechanisms and reduces the selection pressure for resistance development.
3Productivity
If a chimeric enzyme combining LytM and lysostaphin domains is created, then bacteriolytic activity is enhanced, but the complexity of the protein structure increases
Solution Approach 1:
The invention divides the functional requirements of the enzyme into distinct domains: the LytM catalytic domain (residues 185-316) provides the bacteriolytic activity through peptidoglycan hydrolysis, while the lysostaphin cell wall binding domain provides specific targeting to staphylococcal cell walls. This segmentation of functions into modular domains allows each part to perform its specialized role efficiently, with the peptide linker facilitating proper spatial arrangement and interaction between domains.
4Productivity
If the LytM catalytic domain is used alone, then enzymatic activity is present, but the enzyme lacks specificity for bacterial cell wall targeting
Solution Approach 1:
The lysostaphin cell wall binding domain serves as an intermediary that mediates the interaction between the chimeric enzyme and the bacterial cell wall. This binding domain specifically recognizes and attaches to components of the staphylococcal cell wall, positioning the LytM catalytic domain in close proximity to its substrate (peptidoglycan cross-links). This intermediary function ensures that the enzymatic activity is directed precisely at the intended target, enhancing both specificity and efficiency.
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 recombinant polypeptide demonstrates rapid and effective bacteriolytic activity against Staphylococcus strains in raw milk and serum, reducing bacterial counts and addressing antibiotic resistance, while being specific to target bacteria to minimize side effects on non-target organisms.
Implementation Method 1
a LytM catalytic domain (LytMCD), comprising an amino acid sequence shown in SEQ ID NO: 1 or having at least 80% identity thereto
Implementation Method 2
a lysostaphin cell wall binding domain (LssCWT), comprising an amino acid sequence shown in SEQ ID NO: 2 or having at least 80% identity thereto
Data Source
AI summary
The present invention relates to a recombinant polypeptide comprising a LytM catalytic domain, and a lysostaphin cell wall binding domain, optionally, an amino acid linker, and, optionally, a domain directing to the inside of an eukaryotic cell, for use as a medicine, antiseptic agent, antibacterial agent and/or anti-inflammatory agent. In particular, the invention relates to a recombinant polypeptide for use in the prevention and/or treatment of a dermatological disorder and/or inflammation. The recombinant polypeptide formulated for topical administration is also encompassed. A veterinary composition for intramammary administration comprising the recombinant polypeptide, in particular, for use in the treatment of mastitis, is also included. The invention also relates to the use of the recombinant polypeptide as a diagnostic reagent, disinfectant, antiseptic and antibacterial agent, as well as a pharmaceutical, cosmetic or care composition comprising thereof.