Peptide Tags Stabilizing Polymerase Activity
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Solution Overview
Problem
There is a need for methods and compositions that enhance the stability of proteins, particularly polymerases like Taq polymerases, to maintain enzymatic activity at temperatures above freezing, and to improve fidelity, sensitivity, and yield.
Innovation Solution
The use of peptides or peptide tags with specific amino acid sequences, such as those shown in SEQ ID NOs: 1, 8, 10, 13, and 14, which are covalently or non-covalently linked to proteins, enhance stability and retain enzymatic activity at temperatures between -20°C and 50°C, inhibiting degradation and maintaining protein function for extended periods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Temperature
If polymerases are exposed to temperatures above freezing for extended periods, then enzymatic activity is maintained, but protein stability and activity are lost due to degradation
Solution Approach 1:
The patent introduces peptide tags as intermediary molecules that bind to polymerases and stabilize their structure. These peptide tags act as mediators between the polymerase and the thermal environment, protecting the enzyme from degradation at elevated temperatures while maintaining catalytic activity. The peptide tags form a protective complex with the polymerase, allowing storage at higher temperatures without loss of function.
2Stability of the object's composition
If peptide tags are added to enhance stability, then protein stability increases, but device complexity increases due to additional components
Solution Approach 1:
The patent modifies the polymerase by adding peptide tags with specific amino acid sequences (such as those shown in SEQ ID NOs: 1, 8, 10, 13, and 14). This parameter change in the protein structure fundamentally alters its thermal stability properties. The peptide tags change the physical-chemical parameters of the polymerase, enabling it to maintain stability at higher temperatures without requiring complex external stabilization systems.
3Ease of operation
If polymerases are stored at room temperature for convenience, then ease of operation improves, but enzymatic activity decreases due to degradation
Solution Approach 1:
The peptide-tagged polymerases are self-stabilizing and do not require external protection or specialized storage conditions. The peptide tags enable the enzymes to autonomously maintain their structure and activity at room temperature, eliminating the need for refrigerated storage. This self-service capability allows convenient storage and handling while preserving full enzymatic functionality.
Data Source
AI summary
This disclosure provides peptides, polypeptides, fusion polypeptides, compositions, and methods for enhancing or increasing the stability of a polypeptide (e.g., Taq polymerase). Such peptides, polypeptides, fusion polypeptides, or compositions include polypeptides linked to a peptide tag that enhances the stability of the polypeptide. The peptides, polypeptides, fusion polypeptides, compositions may also enhance the activity, specificity, and/or fidelity of other polypeptides in a reaction mixture. The disclosure also provides methods of using such peptides, polypeptides, fusion polypeptides, compositions.