Ancestral Lipase Screening for Surfactant-Resistant Detergency
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Solution Overview
Problem
Existing lipases used in cleansing compositions face challenges with activity inhibition by surfactants and low detergency, hindering their effectiveness in various applications.
Innovation Solution
A method involving ancestral sequence reconstruction is employed to design lipase ancestral sequences, which are then heterologously expressed and tested for resistance to surfactant inhibition and detergency, focusing on the QGLP and Proteus/Yersinia clades.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional lipases are used in cleansing compositions, then they can perform basic lipolytic function, but their activity is inhibited by surfactants and they show low detergency
Solution Approach 1:
The invention uses ancestral sequence reconstruction to design lipase variants with modified amino acid sequences, thereby changing the structural parameters of the enzyme to improve its resistance to surfactant inhibition while maintaining lipolytic activity
Solution Approach 2:
The invention performs preliminary screening of ancestral lipase sequences through in silico design and in vitro validation before industrial application, identifying variants with improved surfactant resistance in advance of large-scale production
2Ease of manufacture
If bacterial lipases from subfamily I.1 or I.2 are produced by heterologous expression, then they can be manufactured, but they require specific chaperones which increases production cost
Solution Approach 1:
The invention extracts and focuses on the specific clade Proteus/Yersinia within subfamily I.1, selecting lipase sequences from this particular group that inherently do not require chaperones for proper folding and activity
Solution Approach 2:
The invention selects lipase variants that can be produced without expensive chaperone proteins, using simple, cost-effective expression systems that do not require additional complex components
3Reliability
If lipases are screened for surfactant resistance, then improved cleansing performance can be achieved, but traditional screening methods are time-consuming and resource-intensive
Solution Approach 1:
The invention performs preliminary in silico analysis and ancestral sequence design to pre-identify promising lipase variants before conducting wet-lab validation, significantly reducing the number of candidates that require experimental testing
Solution Approach 2:
The invention focuses screening efforts on specific clades (Proteus/Yersinia) with known advantages, rather than screening all possible lipase sequences, thereby achieving efficient identification of suitable variants with reduced resources
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 method efficiently identifies lipases with enhanced resistance to surfactant inhibition and improved detergency, outperforming existing lipases in cleansing environments.
Implementation Method 1
Lipases hydrolyze triglycerides to produce fatty acids, thereby contributing to the removal of stains including oil
Data Source
AI summary
Provided is a lipase search method which can efficiently search for lipases having resistance to activity inhibition by a surfactant and/or showing excellent detergency in the presence of a surfactant. A lipase search method comprising steps of: designing lipase ancestral sequences by ancestral sequence reconstruction using a lipase sequence group; measuring at least one of a lipase activity of lipases consisting of the designed ancestral sequences in the presence of a surfactant and a detergency thereof in the presence of a surfactant; and comparing measured results with at least one of a lipase activity of a standard lipase in the presence of a surfactant and a detergency thereof in the presence of a surfactant.