Protease Stability in Detergents via Boronic Acid Mediation
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Solution Overview
Problem
Protease- and amylase-containing cleaning agents suffer from inadequate storage stability and loss of amylolytic activity due to protease inactivation, leading to suboptimal cleaning performance.
Innovation Solution
Incorporating specific proteases with amino acid substitutions (e.g., R99E, S3T, V4I, V199I) in combination with 4-formylphenylboronic acid and another enzyme, such as amylase, to enhance stability and amylolytic activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If protease is added to cleaning agents to improve cleaning performance, then cleaning efficacy is improved, but amylolytic activity is lost due to protease inactivation of amylase
Solution Approach 1:
4-formylphenylboronic acid serves as an intermediary substance that selectively binds to the protease, modulating its activity to prevent amylase inactivation while preserving the protease's cleaning function. This mediator resolves the conflict between maintaining protease efficacy and protecting amylase activity.
Solution Approach 2:
The invention changes the chemical parameters of the protease through specific amino acid substitutions (R99E, S3T, V4I, V199I) to alter its interaction characteristics. These parameter changes enable the protease to work effectively in the presence of 4-formylphenylboronic acid without inactivating amylase, thus resolving the contradiction.
2Productivity
If protease is included in cleaning agents, then cleaning performance is enhanced, but storage stability deteriorates
Solution Approach 1:
4-formylphenylboronic acid acts as a stabilizing intermediary that forms a complex with the protease during storage, preventing unwanted protease activity and self-degradation. This mediator maintains the protease in a stable state until use, resolving the storage stability issue.
Solution Approach 2:
The protease is pre-modified with specific amino acid substitutions (R99E, S3T, V4I, V199I) before formulation, creating a more stable enzyme variant that is less prone to degradation during storage. This preliminary modification action enhances long-term stability while preserving cleaning performance.
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 combination results in stable cleaning agents with improved enzymatic performance and enhanced cleaning efficacy, maintaining optimal activity during storage and use.
Implementation Method 1
4-formylphenylboronic acid is a protease inhibitor known from the prior art, the inhibiting effect of which is described in the granted European patent EP 832 174 B1
Implementation Method 2
at least one protease from the group comprising an amino acid sequence which corresponds to that shown in SEQ ID NO. 1 given amino acid sequence is at least 95% identical over its entire length and counted according to SEQ ID NO. 1 has the amino acid substitution R99E in combination with S3T, V4I and V199I
Data Source
AI summary
The invention relates to cleaning agents containing a) at least one peptidase from the group a1) peptidases, said peptidase comprising an amino acid sequence which over its entire length is at least 80% identical to the amino acid sequence given in SEQ ID NO. 1 and which in the count according to SEQ ID NO. 1 has the amino acid substitution R99E combined with at least two additional amino acid substitutions selected from the group consisting of S3T, V4I and V199I; a2) a peptidase comprising an amino acid sequence which over its entire length is at least 80% identical to the amino acid sequence given in SEQ ID NO. 2 and which in the count according to SEQ ID NO. 2 has the amino acid substitution S99A combined with an insertion between positions 99 and 100, the inserted amino acid being Asp (D); b) 4-formylphenyl boronic acid and c) at least one additional enzyme which differs from the peptidase a). Said cleaning agents are characterised by an improved enzymatic cleaning performance.