Purified Protease Complex for Animal Feed via Isoelectric Focusing
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Solution Overview
Problem
Existing enzymatic complexes from Streptomyces fradiae for animal nutrition and therapeutic applications are not completely free of biological impurities, which can affect their activity and efficacy, and lack precise characterization and separation of proteases based on their isoelectric points.
Innovation Solution
A purified enzymatic complex is developed, comprising proteases with isoelectric points of approximately 7.0 and 8.0, obtained through fermentation, ultrafiltration, ion exchange chromatography, and isoelectrofocusing, with a specific activity of 150,000 and 38,000 Anson units/mg of proteins respectively, and a process that ensures the removal of filler substances devoid of proteolytic activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing enzymatic complexes from Streptomyces fradiae are used for animal nutrition, then proteolytic activity is provided, but biological impurities remain that affect activity and efficacy
Solution Approach 1:
The complex proteolytic activity is segmented into distinct proteases with different isoelectric points (pI around 7.0 and pI around 8.0). This segmentation allows selective purification of individual protease types through isoelectric focusing, separating them from biological impurities while maintaining their beneficial proteolytic functions in animal nutrition
Solution Approach 2:
The patent extracts and isolates specific proteases from the Streptomyces fradiae fermentation broth using a multi-step purification process including ultrafiltration, ion exchange chromatography, and isoelectric focusing. This extraction removes biological impurities while concentrating the active proteolytic enzymes for reliable use in animal feed
2Measurement precision
If proteases are not separated and characterized by isoelectric point, then production is simpler, but precise characterization and control of enzyme activity is lost
Solution Approach 1:
The patent utilizes the isoelectric point (pI) parameter as a key characteristic to differentiate and purify specific proteases from Streptomyces fradiae. By controlling pH during isoelectric focusing, the method achieves precise separation and characterization of proteases with pI around 7.0 and pI around 8.0, enabling accurate measurement and control of enzyme activity despite the added purification steps
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 complex provides precise therapeutic effects, improves animal health and growth, reduces food consumption, and increases profitability in industrial farming by ensuring a technically defined and active product with enhanced proteolytic activity and reduced contamination.
Implementation Method 1
The strain of Streptomyces fradiae can produce at least five types of proteases designated Ia, Ib, II, III, IV and two peptidases
Implementation Method 2
extraction of the complex by filtration, then ultrafiltration on a membrane having a cut-off threshold corresponding to a molecular weight of between 2000 and 12,000
Implementation Method 3
ion exchange chromatography followed by isoelectrofocusing
Implementation Method 4
among these proteases in the mixture one of them has an isoelectric point of around 7.0 and a specific activity of approximately 150,000 units Anson / mg of proteins and another protease has an isoelectric point of the order of 8.0
Implementation Method 5
finally the complex obtained is lyophilized
Data Source
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AI summary
Supplementing farm animal feed, involves adding an enzymatic complex comprising a mixture of proteases obtained by culturing a Streptomyces fradiae strain to farm animal feed, where among this mixture of proteases one of them has an isoelectric point of 7 and another has an isoelectric point of 8. Independent claims are included for 1) feed composition for farm animals comprising: the enzymatic complex comprising a mixture of proteases obtained by culturing a Streptomyces fradiae strain, and optionally including excipients or vehicles for nutriments; and 2) manufacturing the enzymatic complex involving: culturing the Streptomyces fradiae strain, filtering the fermentation broth, then extracting the enzymatic complex by ultrafiltration and ion exchange chromatography followed by an isoelectric focusing, and finally, lyophilizing the complex thus obtained.