Hybrid Endolysin Polypeptide for Gram-Negative Bacterial Infection Control
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Solution Overview
Problem
Current antimicrobial agents are ineffective against Gram-negative bacteria due to their protective outer membrane, which endolysins cannot penetrate, making it challenging to develop effective therapies for multidrug-resistant Gram-negative pathogens.
Innovation Solution
A novel polypeptide with endolysin activity, isolated from Citrobacter koseri phage CkP1, is developed, which has peptidoglycan-degrading capabilities and can target Gram-negative bacteria, potentially overcoming the barrier of the outer membrane by incorporating a peptide stretch with membrane-disrupting activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If endolysins are used as antimicrobial agents, then they can degrade bacterial peptidoglycan effectively, but they cannot penetrate the outer membrane of Gram-negative bacteria
Solution Approach 1:
The patent combines two distinct functional components into a single hybrid polypeptide: an endolysin domain for peptidoglycan degradation and a membrane-active peptide domain for outer membrane disruption. This merging allows the single molecule to overcome both the outer membrane barrier and degrade the peptidoglycan layer, resolving the contradiction between effective antimicrobial action and membrane penetration capability
Solution Approach 2:
The invention creates a composite polypeptide structure integrating two different functional domains with distinct mechanisms of action. The endolysin portion provides enzymatic peptidoglycan hydrolysis while the membrane-active peptide portion provides physical disruption of the outer membrane, creating a composite agent that addresses both barriers presented by Gram-negative bacteria
2Reliability
If conventional antibiotics are used, then they can treat Gram-positive bacterial infections, but they are ineffective against multidrug-resistant Gram-negative bacteria
Solution Approach 1:
The patent fundamentally changes the mechanism of action parameter from conventional antibiotic approaches (which target various cellular processes) to a dual-action approach combining enzymatic peptidoglycan degradation with physical membrane disruption. This parameter change enables activity against Gram-negative bacteria including multidrug-resistant strains that have developed resistance to traditional antibiotics
3Stability of the object's composition
If the outer membrane integrity is maintained through LPS and lipid A interactions, then bacterial stability is preserved, but antimicrobial penetration is blocked
Solution Approach 1:
The membrane-active peptide portion of the hybrid polypeptide acts as an intermediary that specifically targets and disrupts the LPS-lipid A interactions maintaining outer membrane integrity. By inserting into and disrupting this structured barrier, the intermediary peptide enables subsequent access of the endolysin to the peptidoglycan layer without completely compromising bacterial stability until the desired disruptive effect is achieved
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 polypeptide demonstrates significant antibacterial activity against various Gram-negative pathogens, including multidrug-resistant strains, offering a promising alternative therapy for infections and contamination control.
Implementation Method 1
Endolysins are specialized peptidoglycan-degrading enzymes
Implementation Method 2
The LPS cation-binding sites (ionic interactions)
Implementation Method 3
the dense hydrophobic stacking of the Lipid A acyl chain groups (hydrophobic interactions)
Data Source
Figure 1A~1C
Figure 2
Figure 3A~3B
AI summary
The present invention relates to a polypeptide with endolysin activity and amino acid sequences according to SEQ ID No. 1 and fragments or derivatives thereof. Moreover, the present invention relates to nucleic acid molecules encoding said polypeptide, vectors comprising said nucleic acid molecules and host cells comprising either said nucleic acid molecules or said vectors. In addition, the present invention relates to said polypeptide for use as a medicament, in particular for the treatment or prevention of Gram-negative bacterial infections, as diagnostic means, as cosmetic substance or as sanitizing agent. The present invention also relates to the use of said polypeptide for the treatment or prevention of bacterial contamination, particularly of Gram-negative contamination, of foodstuff, of food processing equipment, of food processing plants, of surfaces coming into contact with foodstuff, of medical devices, of surfaces in hospitals and doctor's offices. Furthermore, the present invention relates to a pharmaceutical composition comprising said polypeptide.