Pyolysin Yield Optimization via Chelating Agent Culture
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Solution Overview
Problem
Current methods fail to effectively increase the yield of pyolysin produced by Trueperella pyogenes, a bacterium responsible for bovine metritis, which is crucial for vaccine development and treatment of the condition.
Innovation Solution
A method involving a basal medium with specific carbon sources, chelating agents like EGTA, controlled pH, temperature, and supplementation with vitamins and hemin to enhance pyolysin production, followed by purification and inactivation for vaccine development.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional culture methods are used, then T. pyogenes grows normally, but pyolysin yield remains low
Solution Approach 1:
The patent applies parameter changes by modifying culture conditions including adding chelating agents (EDTA, EGTA) at specific concentrations (0.1-10 mM), controlling pH (6.0-8.0), temperature (28-32°C), and using specific carbon sources (glucose, galactose, sucrose, maltose, oligosaccharides, glycerol, lactose, dextran, dextrin, mono methyl succinate, or N-acetyl glucosamine) to maximize pyolysin production while maintaining manufacturability
Solution Approach 2:
The patent applies preliminary action by adding chelating agents to the medium before the exhaustion of glucose, which prepares the culture environment in advance to optimize pyolysin production during the critical growth phase, thereby increasing yield without complicating the manufacturing process
2Reliability
If pyolysin is produced in high yield, then vaccine effectiveness improves, but production cost increases
Solution Approach 1:
The patent optimizes production parameters including chelating agent concentration (0.1-10 mM), pH (6.0-8.0), temperature (28-32°C), and carbon source selection to achieve high pyolysin yields, thereby improving vaccine effectiveness while controlling production costs through efficient parameter optimization rather than expensive materials
Solution Approach 2:
The patent uses readily available and inexpensive materials such as common chelating agents (EDTA, EGTA), standard carbon sources (glucose, galactose, sucrose, maltose, oligosaccharides, glycerol, lactose, dextran, dextrin, mono methyl succinate, or N-acetyl glucosamine), and basic buffer systems that the system itself can utilize, reducing the need for expensive additives while maintaining high pyolysin production for effective vaccines
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 method significantly increases pyolysin yield and allows for the production of immunogenic compositions for effective vaccination against bovine metritis, providing a promising solution for the condition.
Implementation Method 1
adding a chelating agent to the medium prior to the exhaustion of the glucose in the medium
Data Source
AI summary
Methods for improved cultivation media and culture conditions for Trueperella pyogenes are disclosed herein. Also disclosed are improved methods for the isolation and purification of pyolysin from Trueperella pyogenes.