Subtilase Variants for Detergent Stability Under Harsh Wash Conditions
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing proteases used in detergents face challenges with stability, particularly in wash conditions, leading to inactivation due to pH, temperature, and chelation instability, resulting in reduced wash performance.
Innovation Solution
Development of subtilase variants with specific amino acid substitutions, such as X9C, X9D, X9E, X43A, and others, enhancing stability and wash performance compared to parent subtilases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional proteases are used in detergent formulations, then they provide basic washing functionality, but they exhibit insufficient stability and wash performance under varying temperature and pH conditions
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions at defined positions in the subtilase sequence. These substitutions alter the enzyme's physical and chemical properties to enhance stability across varying temperature and pH conditions while maintaining proteolytic activity in detergent formulations.
Solution Approach 2:
The invention applies local quality by making targeted amino acid substitutions at specific positions (e.g., positions 9, 43, 72, 78, 104, 114, 115, 120, 129, 147, 149, 158, 160, 161, 163, 182, 185, 188, 191, 204, 205, 206, 209, 212, 216, 217, 218, 222, 225, 255, 256, 259, 260, 261, 262) rather than throughout the entire sequence. Each substitution is strategically placed to improve local stability characteristics without compromising overall enzyme function.
2Productivity
If proteases are used in wash conditions, then they provide stain removal capability, but they undergo inactivation due to pH, temperature or chelation instability resulting in loss of wash performance
Solution Approach 1:
The patent applies preliminary action by pre-modifying the subtilase sequence through specific amino acid substitutions before the enzyme is exposed to harsh wash conditions. These substitutions are designed to preemptively enhance resistance to pH changes, temperature fluctuations, and chelation, ensuring the enzyme maintains activity throughout the wash cycle.
Solution Approach 2:
The invention applies beforehand cushioning by introducing amino acid substitutions that create a buffer against environmental stressors. The substituted residues provide structural stability and chemical resistance that cushion the enzyme from inactivation during exposure to varying pH, temperature, and chelating agents in detergent formulations.
3Reliability
If amino acid substitutions are introduced to improve stability, then enzyme stability increases by at least 25%, but the complexity of protein engineering increases
Solution Approach 1:
The patent systematically applies parameter changes by defining specific amino acid substitutions at predetermined positions in the subtilase sequence. This structured approach to modifying enzyme parameters balances the complexity of protein engineering with the achievement of at least 25% improved stability, as each substitution is carefully selected based on its impact on structural and functional properties.
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 exhibit at least 25% improved stability and wash performance, maintaining enzymatic activity under varying detergent conditions, thus improving stain removal efficiency.
Implementation Method 1
subtilase variants having protease activity
Data Source
AI summary
The present invention relates to novel subtilase variants exhibiting increased stability and preferably on par or improved wash performance. The variants of the invention are suitable for use in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dish wash compositions, including automatic dish wash compositions. The present invention also relates to isolated DNA sequences encoding the variants, expression vectors, host cells, and methods for producing and using the variants of the invention.