Fermentation Broth Protein Recovery with Divalent-Cation Stabilization
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Solution Overview
Problem
Existing methods for recovering proteins from fermentation broths often result in partial phase transfer from solubilized to insolubilized states, leading to product loss and increased manufacturing costs due to complex separation processes, and can cause loss of protein activity with alkaline or acidic pH treatments.
Innovation Solution
Adding a divalent cation to the fermentation broth and adjusting the pH to between 11 and 12.5 stabilizes the protein, allowing for high recovery rates and retention of activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the protein concentration in fermentation broth is increased to achieve high titers, then productivity is improved, but the protein undergoes partial phase transfer from solubilized to insolubilized state causing product loss and complex separation
Solution Approach 1:
The patent adjusts pH to a specific range (above 11 to at most 12.5) and controls divalent cation concentration to maintain protein solubility at high titers, preventing phase transfer to insolubilized state and reducing product loss during separation
Solution Approach 2:
Divalent cations act as intermediary substances that mediate between high protein concentration and solubility maintenance, preventing protein precipitation and facilitating easier separation while preserving product
2Productivity
If alkaline or acidic pH treatment is applied during recovery to separate protein, then separation efficiency is improved, but protein activity is lost
Solution Approach 1:
The patent optimizes pH to a specific alkaline range (above 11 to at most 12.5) that enables effective separation while preserving protein activity, avoiding the activity loss associated with extreme acidic or highly alkaline conditions
Solution Approach 2:
The patent dynamically adjusts pH and divalent cation concentration to achieve the optimal balance between separation efficiency and protein activity retention, rather than using fixed extreme conditions
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 achieves high protein recovery rates with minimal loss of activity, enabling efficient separation and stabilization of proteins like amylase.
Implementation Method 1
adding a divalent cation to a fermentation broth comprising the protein of interest and adjustment of the pH of the fermentation broth to a pH above 11 leads to a high recovery rate and high remaining activity of the protein of interest
Implementation Method 2
adjustment of the pH of the fermentation broth to a pH above 11 leads to a high recovery rate and high remaining activity of the protein of interest
Data Source
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AI summary
The present invention relates to a method for recovering a protein of interest from a fermentation broth by adding a salt of a divalent cation and increasing the pH.