Fusion Polypeptides Combining Endolysin and Antimicrobial Peptides
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Solution Overview
Problem
There is a need for new antimicrobial agents effective against Enterococcus bacteria, particularly those that are resistant to conventional antibiotics and can operate effectively across a broad pH range to address the increasing antibiotic resistance in hospital environments.
Innovation Solution
Development of polypeptides comprising specific amino acid sequences, including endolysin-derived enzymes combined with antimicrobial peptides, which enhance antibacterial activity and pH tolerance, specifically targeting Enterococcus faecalis and faecium bacteria.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional antibiotics are used to treat Enterococcus bacteria, then bacterial infections can be controlled, but bacterial resistance to antibiotics increases
Solution Approach 1:
The patent creates fusion polypeptides combining endolysin (an enzyme that degrades bacterial cell walls) with antimicrobial peptides (AMPs). This composite structure allows the molecule to simultaneously degrade the cell wall and disrupt membrane integrity, creating a dual-mechanism antimicrobial agent that overcomes single-target resistance mechanisms.
Solution Approach 2:
The invention divides the antimicrobial function into two distinct functional domains within a single polypeptide: the endolysin domain (amino acids 1-157) responsible for peptidoglycan degradation, and the AMP domain (amino acids 158-187) responsible for membrane disruption. This segmentation allows each domain to perform its specific function while working synergistically.
2Reliability
If wildtype endolysin is used as antimicrobial agent, then some bacterial activity is achieved, but activity is limited at physiological pH and against diverse Enterococcus strains
Solution Approach 1:
The fusion polypeptide is designed to be universally effective against multiple Enterococcus strains (E. faecalis, E. faecium, and other species) while maintaining activity across a broad pH range (5.0-9.0). The combination of endolysin and AMP domains provides multi-functional capability that adapts to different bacterial targets and environmental conditions.
Solution Approach 2:
The invention changes the functional parameters of the endolysin by fusing it with an AMP domain, thereby extending the pH activity range from the narrow range of wildtype endolysin to a broad range covering physiological pH (7.4) and extreme pH values. This parameter expansion makes the agent versatile for different application conditions.
3Reliability
If endolysin is used alone, then cell wall degradation occurs, but complete bacterial elimination is limited due to pH sensitivity and resistance mechanisms
Solution Approach 1:
The patent merges two previously separate antimicrobial mechanisms into a single fusion polypeptide: endolysin-mediated cell wall hydrolysis and AMP-mediated membrane disruption. This combination ensures that bacteria cannot survive through single-mechanism resistance, as both cell wall integrity and membrane function must be compromised simultaneously for complete bacterial elimination.
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 polypeptides demonstrate significantly increased antibacterial activity against a diverse set of Enterococcus strains, achieving near-complete bacterial elimination and maintaining effectiveness across a broad pH range, including physiological pH, surpassing the performance of wildtype endolysin.
Implementation Method 1
They are synthesized during late gene expression in the lytic cycle of phage multiplication and mediate the release of progeny virions from infected cells through degradation of the bacterial peptidoglycan. They are either N-acetyl-P-D-muramidases (lysozymes), lytic transglycosylases, N-acetyl-P-D-glucosaminidases, N-acetylmuramoyl-L- alanine amidases or endopeptidases.
Implementation Method 2
wherein the second amino acid sequence is an antimicrobial peptide, amphipathic peptide, cationic peptide, hydrophobic peptide, sushi peptide or defensin
Data Source
AI summary
The present invention relates to the field of antimicrobial agents active against Enterococcus bacteria. In particular, the present invention relates to a polypeptide comprising a first and a second amino acid sequence, wherein the first amino acid sequence is a sequence selected from SEQ ID NO:2; SEQ ID NO:3; SEQ ID NO: 5; and derivatives thereof; and wherein the second amino acid sequence is an antimicrobial peptide, amphipathic peptide, cationic peptide, hydrophobic peptide, sushi peptide or defensin. In addition, the present invention relates to nucleic acids encoding such polypeptides, vectors comprising such nucleic acids, and corresponding host cells. Finally, the present invention relates to applications of the inventive polypeptides, nucleic acids, vectors, and/or host cells, in particular in the pharmaceutical field.