Phytase Polypeptides 119N 120L 121E Stability Activity
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Solution Overview
Problem
Current phytases lack improved specific activity, stability, pH and temperature stability, pH activity profile, temperature activity profile, substrate profile, and performance in animal feed, which are essential for efficient nutrient utilization in animal feed applications.
Innovation Solution
Development of polypeptides with specific amino acid sequences, such as those comprising 119N, 120L, and 121E, which enhance specific activity, stability, and performance by optimizing the active site and structural regions, leading to improved thermostability and pH stability, and improved substrate utilization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional phytase sequences are used, then basic phytase activity is maintained, but specific activity and stability are insufficient
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues at positions 119, 120, and 121 in the phytase sequence. These sequential changes (119N, 120L, 121E) alter the enzyme's kinetic parameters and stability characteristics, achieving both improved specific activity and enhanced stability simultaneously through precise sequence optimization.
Solution Approach 2:
The invention applies local quality by focusing mutations on specific localized regions (residues 119-121) of the phytase protein. By concentrating amino acid changes in this specific local area rather than throughout the entire sequence, the patent achieves targeted optimization of active site properties and stability without compromising the overall enzyme structure.
2Productivity
If phytase activity is increased, then nutrient utilization improves, but pH and temperature stability deteriorate
Solution Approach 1:
The patent resolves this contradiction by changing the biochemical parameters of the enzyme through specific amino acid substitutions. The modified residues at positions 119-121 alter the enzyme's catalytic efficiency and stability parameters, enabling high phytase activity to be achieved while maintaining pH and temperature stability through optimized molecular properties.
3Ease of manufacture
If wild-type phytase sequences are used, then production is simple, but performance in animal feed is insufficient
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid modifications (119N, 120L, 121E) to the phytase sequence. These changes optimize the enzyme's performance in animal feed applications while maintaining a relatively simple production process, as the modifications are made to a well-characterized base sequence that can be produced using standard recombinant DNA techniques.
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 optimized polypeptides demonstrate enhanced specific activity, stability, and performance in animal feed, effectively improving nutrient availability and utilization, thereby improving animal feed efficiency.
Implementation Method 1
The present invention relates to isolated polypeptides having phytase activity... phytase activity... catalyzes the hydrolysis of phytate
Data Source
AI summary
The present invention relates to polypeptides having phytase activity. These polypeptides have an amino acid sequence which has at least 70% identity to either of three phytases derived from the bacterium Buttiauxella, and which comprises at least one of the following amino acids at the position indicated: 119N, 120L, and/or 121E. These phytases have an improved specific activity. Additional specific amino acid substitutions are also disclosed which characterize and distinguish additional phytases of the invention having improved properties such as temperature and/or pH stability, pH activity profile, temperature activity profile, substrate profile, improved performance in animal feed in vitro or in vivo. The invention also relates to isolated polynucleotides encoding the polypeptides, nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.