Engineered Phytase Polypeptides for Thermostable Liquid Feed
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Solution Overview
Problem
Current phytase products used in animal feed are not robust enough to withstand high temperatures during the pelleting process, especially when applied in liquid form, leading to reduced effectiveness and inconsistent activity, which affects animal performance and environmental phosphorus pollution.
Innovation Solution
Engineered phytase polypeptides with specific amino acid substitutions, such as 30(L, I), 37Y, 45P, 67Y, 89T, 182R, 194M, 202S, 228Y, 256H, 261H, 298V, and 314G, that maintain at least 80% sequence identity to SEQ ID NO:1, exhibit improved thermostability and activity ratio at pH 3.5 versus pH 5.5, enabling their use in liquid form during feed pelleting without significant loss of activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If phytase is applied in liquid form to feed, then even distribution and immediate release are improved, but thermostability during pelleting deteriorates
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the phytase enzyme through site-directed mutagenesis. Specific amino acid residues are substituted to alter the enzyme's thermal stability parameters, allowing it to withstand pelleting temperatures while maintaining activity in liquid form. This directly resolves the contradiction by changing the molecular parameters of the enzyme itself.
Solution Approach 2:
The patent employs local quality by making specific localized substitutions at particular amino acid positions (e.g., positions 30, 37, 45, 67, 89, 182, 194, 202, 228, 256, 261, 298, 314) rather than uniformly modifying the entire enzyme. Each substitution targets specific regions to enhance thermostability while preserving catalytic function, allowing the enzyme to have different properties in different locations.
2Productivity
If high temperature pelleting is used, then feed processing efficiency is improved, but enzyme activity is reduced
Solution Approach 1:
The patent applies preliminary anti-action by pre-modifying the phytase enzyme through genetic engineering to resist thermal denaturation before the pelleting process. The engineered enzyme is designed with enhanced thermostability from the outset, creating a counteracting force against the thermal stress of high-temperature pelleting, thereby maintaining enzyme activity throughout the processing.
Solution Approach 2:
The patent implements preliminary action by incorporating the engineered thermostable phytase into the feed formulation before pelleting. The enzyme is prepared in advance with modified amino acid sequences that confer thermal resistance, so it is already primed and protected before exposure to high temperatures during the pelleting process.
3Reliability
If protective coatings are applied to phytase, then thermostability is improved, but device complexity and cost increase
Solution Approach 1:
The patent applies the taking out principle by removing the need for external protective coatings through internal genetic modification. Instead of adding a separate protective layer to the enzyme, the solution extracts the problem by directly engineering the enzyme's amino acid sequence to inherently resist thermal degradation, eliminating the complexity of coating applications.
Solution Approach 2:
The patent implements self-service by enabling the phytase enzyme to protect itself through genetic engineering. The engineered amino acid substitutions give the enzyme intrinsic thermostability, allowing it to withstand pelleting temperatures without requiring external protective measures or coatings, thus simplifying the overall system.
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 engineered phytase polypeptides ensure consistent enzyme activity and improved animal performance by maintaining stability and activity under high-temperature pelleting conditions, reducing phosphorus pollution, and enhancing nutrient availability in animal feed.
Implementation Method 1
Through the action of phytase, phytate is generally hydrolysed to give lower inositol-phosphates and inorganic phosphate.
Data Source
AI summary
Provided herein, inter alia, are engineered phytase polypeptides and fragments thereof with improved thermotolerance as well as methods for producing and using the same for enhancing animal performance on one or more metrics.