Recombinant Subtilisin Protease Stability in Basic pH

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Solution Overview

Problem

There is a need for further serine proteases suitable for specific conditions and uses, particularly in cleaning fabrics and hard surfaces, and industrial applications.

Innovation Solution

Recombinant serine proteases cloned from Bacillus spp., specifically the BspAP02013-clade of subtilisins, which include specific amino acid motifs and have at least 80% amino acid sequence identity to certain sequences, are used in compositions suitable for cleaning and industrial applications.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional serine proteases are used, then general protease activity is achieved, but performance under specific conditions (basic pH, elevated temperatures) is insufficient

Engineering Contradiction:

Improveprotease activity under basic conditions and elevated temperaturesVSAvoidsuitability for specific cleaning applications

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent applies local quality by introducing specific amino acid substitutions at particular positions (e.g., F23Y, L85F, L120F, S175Y) within the subtilisin structure to enhance stability and activity under basic conditions and elevated temperatures, while maintaining the overall protease function

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent changes the biochemical parameters of the protease by modifying amino acid sequences to achieve optimal performance at basic pH (9-11) and elevated temperatures (50-70°C), transforming the enzyme's characteristics to suit specific cleaning applications

Inventive Principle:

Principle #35Parameter changes

2Productivity

If existing subtilisin variants are used, then some protease activity is achieved, but cleaning effectiveness on fabrics and hard surfaces is insufficient

Engineering Contradiction:

Improvecleaning effectivenessVSAvoidenzyme structure complexity

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent optimizes cleaning effectiveness by modifying specific parameters of the subtilisin structure, including amino acid substitutions at key positions (F23, L85, L120, S175) and adjusting molecular weight through targeted deletions, thereby enhancing cleaning performance without excessive structural complexity

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent creates composite enzyme structures by combining stabilized subtilisin variants with detergent formulations containing surfactants, builders, and other cleaning agents, achieving synergistic cleaning effects on both fabrics and hard surfaces

Inventive Principle:

Principle #40Composite materials

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 recombinant serine proteases demonstrate protease activity in basic conditions and at elevated temperatures, making them effective for use in cleaning compositions, such as detergents, and various industrial processes.

Implementation Method 1

Serine proteases are enzymes (EC No. 3.4.21) possessing an active site serine that initiates hydrolysis of peptide bonds of proteins

Methodology Applied

Scientific EffectEnzymatic hydrolysis: Hydrolysis

Data Source

PatentUS20250163398A1Serine proteases

Publication Date: 2025.05.22 DANISCO US INC

AI summary

The present disclosure relates to serine proteases and variants thereof. Compositions containing the serine proteases are suitable for use in cleaning fabrics and hard surfaces, as well as in a variety of industrial applications.