Fermentation Broth DNA Reduction via Heat and Flocculation
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Solution Overview
Problem
Current methods are ineffective in completely removing residual host cell DNA from fermentation broths, leading to environmental concerns and increased viscosity, which complicates the protein production process.
Innovation Solution
A method involving heating the fermentation broth to at least 70°C and adding poly aluminium chloride, followed by flocculation to separate microorganisms, effectively reduces DNA levels below detectable limits.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional DNA removal methods are used, then some DNA is removed, but residual DNA remains above detectable limits
Solution Approach 1:
The patent applies parameter changes by heating the fermentation broth to temperatures between 60-90°C (optimally 70-80°C) for 10-60 minutes. This thermal parameter change denatures DNA and proteins, making DNA susceptible to subsequent enzymatic degradation by DNases, thereby achieving complete DNA removal below detectable limits
Solution Approach 2:
The patent introduces DNases (DNA enzymes) as intermediary agents that specifically degrade DNA. The enzymes act as mediators between the heated broth and complete DNA removal, converting harmful DNA into harmless degradation products that can be easily separated from the protein product
2Ease of manufacture
If DNA is not removed from fermentation broth, then the purification process is simpler, but environmental and health concerns arise
Solution Approach 1:
The patent converts the harmful effect of DNA (environmental and health risks) into a beneficial outcome by using heat treatment followed by DNase digestion. The process transforms DNA from a harmful contaminant into degraded fragments that are safely removed, while the same thermal conditions preserve the stability of the desired protein product
3Productivity
If DNA is present in fermentation broth, then no additional processing is needed, but viscosity increases leading to higher energy demand
Solution Approach 1:
The patent applies preliminary action by removing DNA through heat treatment and enzymatic degradation before the main protein purification and processing steps. This preliminary DNA removal prevents subsequent viscosity increases that would require additional energy for stirring, pumping, and handling during downstream processing
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
This approach significantly decreases DNA content in the fermentation broth to levels below 1 μg/ml, ensuring environmental safety and simplifying the protein recovery process without significant loss of protein stability or activity.
Implementation Method 1
heating the fermentation broth to a temperature of at least 70° C.
Implementation Method 2
adding a poly aluminium chloride to the fermentation broth, and separating the flocculated microorganism from the fermentation broth
Data Source
AI summary
A method for reducing the level of DNA in a fermentation broth is disclosed where the method includes a heating step where the broth is heated to a temperature of at least 70° C.