Recombinant Bacillus Serine Proteases for Alkaline Detergent Stability
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Solution Overview
Problem
There is a need for serine proteases suitable for specific conditions and uses beyond the existing industrial applications, particularly for cleaning fabrics and hard surfaces, which require enzymes with enhanced stability and activity in alkaline conditions and elevated temperatures.
Innovation Solution
Recombinant serine proteases cloned from Bacillus spp., specifically the B. agaradhaerens-clade subtilisin, or its active fragments, with defined motifs and amino acid sequences, are used in detergent compositions, retaining protease activity across a pH range of 6 to 12 and temperatures of 55°C to 80°C, and maintaining activity under stressed conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing serine proteases are used in detergent compositions, then cleaning activity is provided, but stability and activity under alkaline conditions and elevated temperatures is insufficient
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of subtilisin to create variants with improved stability and activity under specific conditions (alkaline pH and elevated temperatures). The variant subtilisins contain specific amino acid substitutions that enhance their performance in detergent compositions, allowing them to maintain proteolytic activity across a broader range of temperatures and pH levels compared to wild-type enzymes.
2Reliability
If serine proteases with enhanced stability are developed, then cleaning performance under stressed conditions improves, but the complexity of enzyme selection and optimization increases
Solution Approach 1:
The patent applies local quality by introducing specific amino acid substitutions at particular positions in the subtilisin sequence (e.g., substitutions at positions 30, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100). These localized modifications target specific regions of the enzyme to enhance stability without requiring complete redesign of the entire protein structure.
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 effective cleaning performance on substrates like egg yolk, blood, and ink, and are stable in detergent compositions, ensuring efficient cleaning across a range of conditions, making them suitable for industrial applications.
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
Data Source
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AI summary
The present disclosure relates to serine proteases cloned from Bacillus spp., 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.