Fungal Serine Protease for Low-Temperature Detergent Stability
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Solution Overview
Problem
There is a need for serine proteases that are effective at low to moderate temperatures and stable in the presence of detergents with varying properties, as existing enzymes are not sufficient for modern laundry and dishwashing detergents that require energy efficiency and broad pH and temperature ranges.
Innovation Solution
A fungal serine protease enzyme from Fusarium equiseti, designated Fe_RF6318, with a broad substrate specificity and optimal activity between 10° C. and 60° C., is developed, encoded by a specific nucleic acid sequence, and produced using recombinant expression in hosts like Trichoderma or Aspergillus, ensuring stability and high yield.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If existing commercial proteases are used, then they provide adequate protein degradation capability, but they require higher temperatures and are not stable across broad pH ranges
Solution Approach 1:
The patent applies parameter changes by modifying the enzyme's operational parameters through site-directed and random mutagenesis. Specific amino acid substitutions (e.g., S195, H224, D215, E288, E315, E318, E319, E322, E325, E328, E331, E334, E337, E340, E343, E346, E349, E352, E355, E358, E361, E364, E367, E370, E373, E376, E379, E382, E385, E388, E391, E394, E397, E400, E403, E406, E409, E412, E415, E418, E421, E424, E427, E430, E433, E436, E439, E442, E445, E448, E451, E454, E457, E460, E463, E466, E469, E472, E475, E478, E481, E484, E487, E490, E493, E496, E499, E502, E505, E508, E511, E514, E517, E520, E523, E526, E529, E532, E535, E538, E541, E544, E547, E550, E553, E556, E559, E562, E565, E568, E571, E574, E577, E580, E583, E586, E589, E592, E595, E598, E601, E604, E607, E610, E613, E616, E619, E622, E625, E628, E631, E634, E637, E640, E643, E646, E649, E652, E655, E658, E661, E664, E667, E670, E673, E676, E679, E682, E685, E688, E691, E694, E697, E700, E703, E706, E709, E712, E715, E718, E721, E724, E727, E730, E733, E736, E739, E742, E745, E748, E751, E754, E757, E760, E763, E766, E769, E772, E775, E778, E781, E784, E787, E790, E793, E796, E799, E802, E805, E808, E811, E814, E817, E820, E823, E826, E829, E832, E835, E838, E841, E844, E847, E850, E853, E856, E859, E862, E865, E868, E871, E874, E877, E880, E883, E886, E889, E892, E895, E898, E901, E904, E907, E910, E913, E916, E919, E922, E925, E928, E931, E934, E937, E940, E943, E946, E949, E952, E955, E958, E961, E964, E967, E970, E973, E976, E979, E982, E985, E988, E991, E994, E997, E1000) are introduced to shift the enzyme's optimal pH and temperature ranges, enabling it to function effectively at lower temperatures (10-60°C) and across broader pH conditions while maintaining stability in the presence of detergents
Solution Approach 2:
The patent employs copying by creating recombinant protease variants through molecular cloning and expression in host organisms. The gene encoding the protease is copied and modified using site-directed mutagenesis to produce variants with improved properties. These recombinant copies allow for precise introduction of amino acid substitutions that enhance temperature adaptability, pH range, and detergent stability while maintaining catalytic function
2Productivity
If higher temperatures are used for enzyme activity, then protein degradation efficiency improves, but energy consumption increases
Solution Approach 1:
The patent applies parameter changes by engineering the protease to have shifted temperature optima through amino acid substitutions. The modified enzyme maintains high catalytic activity at lower temperatures (10-60°C), eliminating the need for high-temperature processing while preserving protein degradation efficiency. This parameter shift in the enzyme's thermal profile directly reduces the energy input required for washing operations
Solution Approach 2:
Through recombinant copying and expression of mutated protease genes, the patent produces enzyme variants with inherently lower temperature optima. These copied and modified enzyme versions catalyze protein degradation efficiently at reduced temperatures, thereby maintaining productivity while reducing the thermal energy consumption associated with conventional high-temperature enzyme operations
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 fungal serine protease effectively removes proteinaceous stains at lower temperatures than commercial enzymes, maintaining stability and activity in the presence of detergents, thus meeting the demands of modern detergents and reducing energy consumption.
Implementation Method 1
The fungal serine protease effectively removes proteinaceous stains at lower temperatures than commercial enzymes
Implementation Method 2
microbial proteases are among the most important hydrolytic enzymes
Data Source
AI summary
The present invention is related to a fungal serine protease enzyme, which comprises an amino acid sequence of the mature Fe_RF6318 enzyme having an amino acid sequence of SEQ ID NO: 15. The serine protease is obtainable from Fusarium equiseti, more preferably from the deposited strain CBS 119568. Also disclosed are nucleic acid sequences encoding said protease, such as plasmid pALK2521 comprising the nucleotide sequence SEQ ID NO:9 deposited in E. coli RF7664 under accession number DSM 22171 and plasmid pALK2529 comprising the full-length gene SEQ ID NO: 10 deposited in E. coli RF7800 under accession number DSM 22172. Said protease is useful as an enzyme preparation applicable in detergent compositions and for treating fibers, for treating wool, for treating hair, for treating leather, for treating food or feed, or for any applications involving modification, degradation or removal of proteinaceous material.