Subtilisin Variant Substitutions for Stability and Soil Removal
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Solution Overview
Problem
Existing subtilisins lack improved stability and soil removal performance for cleaning applications.
Innovation Solution
Development of subtilisin variants with specific amino acid substitutions at position 39 and other positions, enhancing stability and cleaning performance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing subtilisin is used for cleaning applications, then basic proteolytic activity is provided, but stability and soil removal performance are insufficient
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid residues at specific positions in the subtilisin sequence. Multiple variants are created with substitutions at positions such as 39, 215, 226, 266, and 286, along with variations in N-terminal extensions and signal peptides. These parameter changes in the protein structure directly improve both stability and cleaning performance, resolving the technical contradiction between reliability and productivity.
2Reliability
If amino acid substitutions are made to improve stability, then protein structure is modified, but complexity of variant development increases
Solution Approach 1:
The patent applies segmentation by dividing the protein modification strategy into distinct modules: N-terminal extensions (signals, leader sequences), core amino acid position substitutions (at positions 39, 215, 226, 266, 286), and C-terminal variations. This modular approach to variant development systematically manages complexity while enabling comprehensive screening of stability-improving mutations.
Solution Approach 2:
The patent systematically varies parameters including amino acid identity at specific positions, N-terminal extension length and composition, and signal peptide sequences. By changing these discrete parameters in a structured manner, the patent manages development complexity while achieving improved stability across multiple variants.
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 variants demonstrate improved cleaning performance and stability, suitable for use in laundry and dishwashing compositions, offering enhanced cleaning efficacy.
Implementation Method 1
A protease (also known as a proteinase) is an enzyme that has the ability to break down other proteins. A protease has the ability to conduct proteolysis, which begins protein catabolism by hydrolysis of peptide bonds that link amino acids together
Implementation Method 2
Serine proteases are enzymes (EC No. 3.4.21) possessing an active site serine that initiates hydrolysis of peptide bonds of proteins
Data Source
AI summary
Disclosed herein is one or more subtilisin variant, nucleic acid encoding same, and compositions and methods related to the production and use thereof, including one or more subtilisin variant that has improved stability and/or soil removal compared to one or more reference subtilisin.