Bacterial Heme Peroxidase Pathogen Control via Organic Solvent Treatment
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Solution Overview
Problem
Current bacterial heme peroxidases, such as bovine lactoperoxidase and LspPOX, exhibit low enzymatic activity and high production costs, making them inefficient and expensive for large-scale applications in pathogen control.
Innovation Solution
Development of novel bacterial heme peroxidases, specifically HydPOX, HydPOXs, and OkePOXs, which demonstrate significantly increased peroxidase activity compared to existing enzymes, achieved through optimized production in engineered Bacillus subtilis strains.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If bovine lactoperoxidase or LspPOX is used as a biocontrol agent, then pathogen control function is provided, but enzymatic activity is low and production cost is high
Solution Approach 1:
The patent applies parameter changes by modifying the enzymatic properties of peroxidases through the use of organic solvents. Specifically, the peroxidase is treated with organic solvents to alter its conformational state, which increases its enzymatic activity and reduces KM values for substrates like H2O2 and iodide. This allows the enzyme to achieve higher catalytic efficiency and lower production costs while maintaining pathogen control function.
Solution Approach 2:
The patent creates improved copies of existing peroxidases by producing bacterial peroxidases (such as from Bacillus species) that serve as functional alternatives to bovine lactoperoxidase. These bacterial peroxidases are engineered or selected to have inherently higher enzymatic activity and are then further optimized using organic solvent treatment, providing a cost-effective copy with superior performance characteristics.
2Reliability
If bovine lactoperoxidase is used as a biocontrol agent, then pathogen control function is provided, but production cost is prohibitively expensive for large-scale applications
Solution Approach 1:
The patent employs bacterial peroxidases that can be produced more cheaply than bovine lactoperoxidase through microbial fermentation processes. These bacterial enzymes serve as disposable, cost-effective alternatives that can be manufactured at scale without the high costs associated with animal source extraction, purification, and stabilization required for bovine lactoperoxidase.
Solution Approach 2:
By treating peroxidases with organic solvents to change their conformational parameters, the patent achieves enhanced enzymatic activity that reduces the amount of enzyme needed per application. This parameter modification makes the overall production and application process more cost-effective for large-scale agricultural use.
3Reliability
If high concentrations of enzyme are used to achieve protective effects, then pathogen control efficiency is maintained, but production cost increases
Solution Approach 1:
The patent reduces the required enzyme concentration by changing the operational parameters of the peroxidase through organic solvent treatment. This modification increases the enzyme's catalytic efficiency and decreases its KM values, allowing it to achieve the same protective effect at lower concentrations, thereby reducing the quantity of substance needed and lowering production costs.
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 new bacterial heme peroxidases show enhanced enzymatic activity, requiring lower enzyme concentrations to achieve the same protective effects, thus reducing production costs and improving pathogen control efficiency.
Implementation Method 1
Peroxidases are enzymes found in all plants and animals and they are essential for living systems. Depending on the chemistry of the active site, they can be classified in heme and nonheme peroxidases. Most peroxidases are heme containing enzymes catalyzing the oxidation of diverse substrates in the presence of hydrogen peroxide (H2O2).
Implementation Method 2
Most peroxidases are heme containing enzymes catalyzing the oxidation of diverse substrates in the presence of hydrogen peroxide (H2O2).
Data Source
Figure 1~2
Figure 3A~3.1B
Figure 4A~4.1B
AI summary
The present invention relates to means and methods comprising a bacterial heme peroxidase and/or a functional fragment thereof for medical and non-medical use in preventing and/or controlling a pathogen on food, plants, or in or on the human or animal body. The present invention thus relates to novel medical and non-medical uses of a bacterial heme peroxidase and/or a functional fragment thereof for preventing and/or controlling a pathogen on food, plants, or on the human or animal body. Furthermore, the present invention relates to corresponding methods comprising a bacterial heme peroxidase and/or a functional fragment thereof for preventing and/or controlling a pathogen on food, plants or on or in the human or animal body comprising applying a sufficient amount of the bacterial heme peroxidase and/or a functional fragment thereof on said food, plants, or on mucosae, skin, in particular epidermal skin like scalp or facial skin and/or teeth to reduce the number of cells of said pathogen. In addition, the invention relates to specific heme peroxidases that have been identified to have desired properties. The invention also relates to fragments of or small heme peroxidases that have particularly advantageous properties. Moreover, the present invention further relates to corresponding compositions comprising a bacterial heme peroxidase and/or a functional fragment thereof for preventing and/or controlling a pathogen on food, plants, or in or on the human or animal body. Kits comprising components used in the methods and compositions of the invention are also provided herein. Furthermore, provided is the use of a bacterial heme peroxidase and/or of a functional fragment thereof for the preparation of the compositions of the present invention. The present invention finally also relates to a method for producing a bacterial heme peroxidase protein and/or a functional fragment thereof in an engineered Bacillus subtilis strain. The inventive use of the bacterial heme peroxidase of the present invention and/or a functional fragment thereof for preventing and/or controlling a pathogen is, without being limiting, particularly useful in ecologically compatible commercial plant/crop protection campaigns as well as in daily personal care products such as toothpaste for example.