Bacillus gibsonii Subtilisin Variants for Enhanced Stability
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Solution Overview
Problem
There is a need for improved protease variants with enhanced stability and cleaning performance for applications such as dishwashing, as existing subtilisin variants do not meet the required standards for stability and effectiveness in removing protein-based stains.
Innovation Solution
Development of Bacillus gibsonii subtilisin variants with specific amino acid substitutions, such as S039E, S099R, S126A, D127E, and F128G, along with additional substitutions like N074D, N085R, and N253P, which are used to create stable expression vectors for improved proteolytic activity and stability in cleaning compositions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing subtilisin variants are used, then basic proteolytic activity is maintained, but stability and cleaning performance are insufficient
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid positions in the subtilisin sequence (e.g., positions 39, 99, 126, 127, 128, 253) to optimize both stability and cleaning performance. The variant subtilisin B.99 contains specific substitutions (S39E, S99R, S126A, D127E, F128G, N253P) that change the physical-chemical parameters of the enzyme, resulting in improved stability and proteolytic activity compared to the parent enzyme.
2Reliability
If amino acid substitutions are introduced to improve stability, then proteolytic activity may be compromised
Solution Approach 1:
The patent systematically changes amino acid parameters at specific positions to achieve both improved stability and maintained proteolytic activity. The variant subtilisin B.99 contains six specific amino acid substitutions that were optimized to balance structural stability with catalytic function, demonstrating that parameter changes can simultaneously improve both properties rather than trade one for the other.
Solution Approach 2:
The development of subtilisin variants involved iterative testing and optimization where proteolytic activity and stability were measured and used as feedback to refine the amino acid substitutions. This feedback loop allowed identification of the optimal combination of substitutions (S39E, S99R, S126A, D127E, F128G, N253P) that achieve both high stability and high proteolytic activity.
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 Bacillus gibsonii subtilisin variants demonstrate improved stability and cleaning performance, achieving enhanced proteolytic activity and stability in dishwashing applications, effectively removing protein-based stains compared to reference subtilisins.
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
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.