Subtilase Variants with Specific Mutations for Detergent Stability
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Solution Overview
Problem
Current proteases used in detergents face challenges such as instability under varying washing conditions, including temperature and pH fluctuations, leading to reduced wash performance and enzyme inactivation, necessitating the development of more stable and effective protease variants.
Innovation Solution
Development of subtilase variants with specific mutations at positions 9, 43, 76, 131, 158, 161, 194, 206, 209, 212, 216, 259, and 262, which maintain at least 80% sequence identity to a parent subtilase and include amino acids like T, I, D, E, A, and I, enhancing stability and protease activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional proteases are used in detergents, then wash performance is achieved, but enzyme stability under varying washing conditions deteriorates
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid mutations at positions 9, 43, 76, 131, 158, 161, 194, 206, 209, 212, 216, 259, and 262 in the subtilase protein sequence. These parameter changes in the protein's amino acid composition directly improve the enzyme's stability under varying washing conditions while maintaining wash performance.
Solution Approach 2:
The patent creates a composite enzyme variant by combining multiple specific mutations in the subtilase protein structure. The variant comprises at least four of the following amino acids in the indicated positions: T in position 3, I in position 4, D in position 99, E in position 101, A in position 103, I in position 104, S in position 160 and I in position 205. This composite approach results in an enzyme with enhanced stability and activity.
2Productivity
If protease activity is maintained under diverse washing conditions, then wash performance is preserved, but enzyme inactivation increases
Solution Approach 1:
The patent uses parameter changes by modifying specific amino acid residues in the subtilase protein to create variants with improved resistance to inactivation. The mutations at positions 9, 43, 76, 131, 158, 161, 194, 206, 209, 212, 216, 259, and 262 alter the protein's structural and functional properties to maintain protease activity while resisting inactivation under diverse washing conditions.
Solution Approach 2:
The patent applies beforehand cushioning by introducing stabilizing mutations that preemptively protect the enzyme structure against inactivation. The specific amino acid changes create a more robust protein structure that is pre-conditioned to resist denaturing effects from pH, temperature, and other washing conditions before inactivation can occur.
3Duration of action of stationary object
If storage stability is improved in detergent composition, then enzyme lifespan is extended, but protein sequence complexity increases
Solution Approach 1:
The patent applies parameter changes by introducing a limited number of specific amino acid mutations (at positions 9, 43, 76, 131, 158, 161, 194, 206, 209, 212, 216, 259, and 262) rather than completely redesigning the protein sequence. This controlled parameter change approach extends storage stability while keeping the overall protein sequence relatively simple and recognizable as a subtilase variant.
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 modified subtilase variants exhibit improved storage stability and protease activity, maintaining performance across diverse washing conditions, thereby enhancing wash performance and extending enzyme lifespan in detergent compositions.
Implementation Method 1
The terms 'subtilase' and 'protease' may be used interchangeably herein and refer to an enzyme that hydrolyses peptide bonds in proteins
Data Source
AI summary
The invention relates to subtilase variants having improved stability, compositions comprising the variants, in particular detergent compositions, polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; as well as methods of producing the variants and methods for stabilizing a subtilase variant