Thermally Stable Phospholipase C for High-Temperature Oil Degumming
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Solution Overview
Problem
The thermal stability of existing phospholipase C enzymes is poor, limiting their use in degumming processes to temperatures below 60°C, which restricts efficiency and increases energy consumption.
Innovation Solution
A polypeptide with enhanced phospholipase C activity is developed, featuring specific amino acid substitutions at positions 6, 8, 10, 104, and 205, which significantly improves its thermal stability, allowing for higher temperature degumming.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Temperature
If traditional phospholipase C is used for degumming, then the enzyme can perform catalysis at lower temperatures, but the thermal stability is poor and cannot tolerate temperatures above 60°C
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of phospholipase C through site-directed mutagenesis. Specific amino acid residues are substituted to alter the enzyme's thermal stability parameters, enabling it to maintain catalytic activity at higher temperatures (65-75°C) while preserving its phospholipase C function. This directly resolves the contradiction between operating temperature and thermal stability.
2Productivity
If degumming is performed at higher temperatures to improve oil yield and separation, then the efficiency increases, but existing phospholipase C cannot function above 60°C
Solution Approach 1:
The patent changes the temperature parameter at which the enzyme operates by engineering its thermal stability. The modified phospholipase C maintains catalytic activity at 65-75°C, enabling high-temperature degumming that improves oil yield and separation efficiency while the enzyme remains functional. This resolves the contradiction between productivity and enzyme reliability at high temperatures.
3Reliability
If the optimal reaction temperature of PLC is lower than 50°C, then the enzyme maintains stability, but cold water is needed to cool crude oil resulting in large energy consumption
Solution Approach 1:
The patent inverts the conventional approach by raising the enzyme's optimal temperature range to 65-75°C instead of operating at lower temperatures and cooling the crude oil. This allows the degumming process to proceed at temperatures compatible with crude oil storage conditions, eliminating or reducing the need for energy-consuming cooling steps while maintaining enzyme stability and activity.
4Productivity
If PLC and PLA1 are used in combination for deep degumming, then the degumming effectiveness improves, but a heat-resistant PLC is needed to match the PLA1 optimal temperature of 65°C
Solution Approach 1:
The patent changes the temperature parameter of PLC to match PLA1's optimal operating temperature of 65°C. By engineering phospholipase C to be stable and active at 65-75°C, the patent enables effective combination of PLC and PLA1 for deep degumming, achieving synergistic effects while maintaining temperature compatibility between the two enzymes.
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 improved thermal stability of the polypeptide enables degumming at higher temperatures, enhancing oil yield, reducing energy consumption, and facilitating its use in combination with other enzymes like PLA1 for deep degumming.
Implementation Method 1
A polypeptide with enhanced phospholipase C activity is developed, featuring specific amino acid substitutions at positions 6, 8, 10, 104, and 205
Implementation Method 2
Phospholipase C (PLC) exhibits significant advantages in, such as increasing yield of diacylglycerols (DAG)
Data Source
AI summary
Provided is a polypeptide having phospholipase C activity. The polypeptide has: 1) an amino acid sequence represented by SEQ ID No: 2, comprising amino acid substitutions occurred at one or more positions, wherein the one or more positions are selected from positions 6, 8, 10, 104, and 205 in the amino acid sequence represented by SEQ ID No: 2 or any combination thereof; or 2) having at least 80% sequence identity with 1), and at least one of positions 6, 8, 10, 104 and 205 being different from positions 6, 8, 10, 104, and 205 in the amino acid sequence represented by SEQ ID No: 2. Provided are a nucleic acid molecule for encoding the polypeptide, a vector comprising the nucleic acid molecule, and a cell comprising the nucleic acid molecule or the vector. Provided are uses of the polypeptide, the nucleic acid molecule, the vector, and the cell.