Fungal Serine Protease for Low-Temperature Detergent Cleaning

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Solution Overview

Problem

Current detergent enzymes, particularly proteases, are ineffective at low temperatures and lack stability in varying detergent conditions, making them unsuitable for modern washing habits and environmental regulations which demand energy efficiency and compatibility with synthetic fibers.

Innovation Solution

A fungal serine protease enzyme with broad substrate specificity and stability across a wide pH and temperature range, specifically designed for use in laundry and dish detergents, is developed. This enzyme is produced using a recombinant expression system in hosts like Trichoderma or Aspergillus and is capable of degrading proteinaceous stains at temperatures as low as 10°C, maintaining activity in the presence of detergents.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If current detergent enzymes (particularly proteases) are used, then they can degrade proteinaceous stains, but they are ineffective at low temperatures and lack stability in varying detergent conditions

Engineering Contradiction:
Improveenzyme effectivenessVSAvoidtemperature range and detergent compatibility
Core Design Contradiction:
ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent applies parameter changes by modifying the enzyme's structural and functional parameters through site-directed and random mutagenesis. Specific amino acid substitutions (e.g., Phe314→Ser, Phe314→Thr, Phe314→Ala, Phe314→Gly, Phe314→Val, Phe314→Leu, Phe314→Ile, Phe314→Met, Phe314→Asp, Phe314→Asn, Phe314→Gln, Phe314→Cys, Phe314→His, Phe314→Arg, Phe314→Lys) were introduced to alter the enzyme's temperature profile and detergent stability while maintaining proteolytic activity. This enabled the enzyme to function effectively at low temperatures (10-40°C) and remain stable in varying detergent conditions.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite enzyme system by combining multiple mutated variants of serine protease in a single formulation. The enzyme composition includes at least two different serine protease variants with complementary properties, where one variant may provide optimal activity at certain temperatures while another provides stability in specific detergent conditions. This composite approach allows the detergent composition to maintain reliable stain degradation across a broad range of temperatures and detergent formulations.

Inventive Principle:
Principle #40Composite materials

2Productivity

If high washing temperatures are used to improve enzyme performance, then proteinaceous stains are removed more effectively, but energy consumption increases and synthetic fibers cannot tolerate the heat

Engineering Contradiction:
Improvestain removal efficiencyVSAvoidwashing energy consumption
Core Design Contradiction:
ProductivityVSUse of energy by moving object

Solution Approach 1:

The patent fundamentally changes the temperature-parameter relationship of the enzyme by introducing mutations that shift the activity profile toward lower temperatures. The mutated serine proteases exhibit peak activity in the 10-40°C range, whereas conventional proteases require 50-60°C or higher for optimal performance. This parameter shift enables effective stain removal at low temperatures, reducing washing energy consumption by 20-40% compared to conventional processes while maintaining or improving productivity.

Inventive Principle:
Principle #35Parameter changes

3Temperature

If conventional proteases are used, then they work at higher temperatures, but they lack stability towards oxidizing agents and changing washing conditions

Engineering Contradiction:
Improveoptimal working temperatureVSAvoidstability towards oxidizing agents and washing conditions
Core Design Contradiction:
TemperatureVSReliability

Solution Approach 1:

The patent applies parameter changes by modifying the enzyme's structural parameters through multiple amino acid substitutions that enhance conformational stability. The mutated variants show improved resistance to oxidizing agents (peroxides, hypochlorite) and maintaining structural integrity across pH 6-11 and temperature ranges of 10-60°C. The combined mutations create a more robust enzyme structure that resists denaturation and degradation under varying washing conditions, thereby improving reliability.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent employs a composite enzyme system where multiple protease variants with different stability profiles are combined. One variant may provide superior resistance to oxidizing agents while another maintains stability across broader pH and temperature ranges. This composite formulation ensures that at least one active variant remains functional under any given washing condition, significantly improving overall reliability and performance consistency.

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 fungal serine protease effectively removes proteinaceous stains at lower temperatures than commercial enzymes, saving energy and maintaining performance in diverse detergent conditions, thus addressing the need for energy-efficient and environmentally friendly cleaning solutions.

Implementation Method 1

Microbial proteases are among the most important hydrolytic enzymes and find applications in various industrial sectors

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentEP2424993B1A fungal serine protease and use thereof
Publication Date: 2013.04.03 AB ENZYMES OY
  • EP2424993B1 patent drawingFigure 1A
  • EP2424993B1 patent drawingFigure 1B
  • EP2424993B1 patent drawingFigure 2

AI summary

The present invention is related to a fungal serine protease enzyme, which comprises an amino acid sequence the mature Fa_RF7182 enzyme having an amino acid sequence of SEQ ID NO: 18. The serine protease is obtainable from Fusarium acuminatum, more preferably from the deposited strain CBS 124084. Also disclosed are nucleic acid sequences encoding said protease, such as plasmid pALK2530 comprising the nucleotide sequence SEQ ID NO: 12 deposited in Escherichia coli RF7803 under accession number DSM 22208 and plasmid pALK2531 comprising the full-length gene SEQ ID NO: 13 deposited in E. coli RF7879 under accession number DSM 22209, as well as fungal hosts, such as Trichoderma. 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.