Engineered Lactase Enzymes for Acidic and High-Temperature Hydrolysis
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Solution Overview
Problem
Existing lactases are not suitable for lactose hydrolysis at high temperatures or low pH values, requiring separate steps and high enzyme dosages, which can lead to microbial growth and are not compatible with ultra-heat treated milk production.
Innovation Solution
Development of beta-galactosidases with enhanced stability and activity at a broad range of temperatures and pH values, allowing for efficient lactose hydrolysis in dairy products, including high-temperature processes and ultra-heat treated milk.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If neutral lactases (pH optimum 6-8, temperature optimum 37°C) are used in low-lactose yoghurt production, then enzyme activity is maintained at neutral pH, but the activity drops as pH decreases during fermentation requiring high enzyme dosages or separate treatment steps
Solution Approach 1:
The patent applies parameter changes by modifying the enzyme's pH and temperature optima through protein engineering. The engineered beta-galactosidase achieves pH optimum around 4.5-5.5 and temperature optimum around 45-55°C, allowing it to maintain high activity during fermentation at acidic pH and elevated temperatures, thereby eliminating the need for separate treatment steps or high enzyme dosages
Solution Approach 2:
The patent implements dynamics by creating an enzyme that adapts to changing process conditions. The engineered lactase maintains stability and activity across a broad range of pH (3-9) and temperature (0-70°C) conditions, allowing the same enzyme to function effectively throughout the fermentation process as pH decreases, rather than requiring process adjustments
2Productivity
If prolonged incubation (10-48 h) at low temperature (6°C) is used with existing lactases, then lactose hydrolysis is achieved, but the process is time-consuming and requires separate steps before fermentation
Solution Approach 1:
The patent applies parameter changes by shifting the temperature optimum to 45-55°C, which accelerates the reaction kinetics significantly. This allows lactose hydrolysis to be completed in hours rather than days, enabling the process to be integrated within normal fermentation timelines without requiring prolonged pre-incubation steps
3Productivity
If high enzyme dosages are used to compensate for activity drop at low pH, then sufficient lactose hydrolysis is achieved, but cost increases and microbial growth risk increases
Solution Approach 1:
The patent applies parameter changes by engineering the enzyme to maintain high specific activity at acidic pH (optimum 4.5-5.5). This allows using lower enzyme dosages while achieving the same hydrolysis rate, thereby reducing the risk of microbial growth from high protein concentrations and lowering enzyme costs
4Object-affected harmful factors
If existing lactases are used for high temperature processing, then microbial count is kept low, but the known lactases are not stable or active at high temperatures
Solution Approach 1:
The patent applies parameter changes by engineering thermophilic stability into the beta-galactosidase, with temperature optimum shifted to 45-55°C and stability maintained up to 70°C. This allows the enzyme to withstand high-temperature pasteurization and UHT processing while remaining active, enabling integration of lactose hydrolysis with high-temperature processing steps
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
Enables the production of lactose-free or low-lactose dairy products with improved efficiency, stability, and reduced microbial growth, using lower enzyme dosages and enabling applications in diverse dairy processes.
Implementation Method 1
Lactase (beta-galactosidase; EC 3.2.1.23) is the enzyme that performs the hydrolysis step of the milk sugar lactose into monosaccharides
Data Source
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AI summary
The present invention relates to new improved peptide or dimeric peptides exhibiting beta-galactosidase enzyme activity as well as improved methods for reducing the lactose content in compositions optionally at elevated temperatures.