Apoplast Protein Recovery by Buffer Infiltration Without Grinding
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Solution Overview
Problem
Conventional methods for recovering target proteins from plant bodies involve grinding, which results in the inclusion of plant-derived insoluble foreign matter and impurities, increasing the load on purification processes.
Innovation Solution
A method involving the injection and extraction of a buffer through stomata, water pores, or cross-sections of a plant body to recover target proteins expressed in the apoplast without grinding, utilizing techniques such as pressurization, depressurization, and osmotic pressure differences.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If grinding of plant body is performed to extract target protein, then the amount of target protein extract can be increased, but the amount of unwanted plant components increases and purification load increases
Solution Approach 1:
The invention extracts only the target protein from the plant body by injecting buffer through stomata to retrieve apoplastic fluid, leaving unwanted plant components (cell walls, organelles, insoluble matter) in the plant tissue. This selective extraction eliminates the need for complex purification steps while maintaining high target protein yield.
2Quantity of substance
If grinding of plant body is performed to extract target protein, then the amount of target protein extract can be increased, but insoluble impurities and plant residues are included in the extract
Solution Approach 1:
The invention extracts only the target protein from the plant body by injecting buffer through stomata to retrieve apoplastic fluid, leaving unwanted plant components (cell walls, organelles, insoluble matter) in the plant tissue. This selective extraction eliminates the need for complex purification steps while maintaining high target protein yield.
3Quantity of substance
If conventional extraction method is used, then target protein can be recovered, but time and energy are consumed in grinding and purification processes
Solution Approach 1:
The invention extracts only the target protein from the plant body by injecting buffer through stomata to retrieve apoplastic fluid, leaving unwanted plant components (cell walls, organelles, insoluble matter) in the plant tissue. This selective extraction eliminates the need for complex purification steps while maintaining high target protein yield.
Solution Approach 2:
The invention replaces the mechanical grinding process with a physiological extraction method using buffer injection through stomata. This substitution eliminates time-consuming mechanical disruption and extensive purification steps, significantly reducing processing time while maintaining efficient target protein recovery.
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
Reduces the need for grinding, thereby minimizing the amount of unwanted plant components, simplifying purification and enabling efficient recovery of target proteins in quantities equivalent to or greater than conventional methods.
Implementation Method 1
injecting the buffer by pressurization or depressurization
Implementation Method 2
injecting the buffer by pressurization or depressurization
Implementation Method 3
extracting the buffer by pressurization or depressurization
Implementation Method 4
extracting the buffer by pressurization or depressurization
Implementation Method 5
immersing the plant body in a buffer having higher osmotic pressure than the buffer having been injected into the plant body
Data Source
AI summary
The present invention discloses a method for recovering a target protein expressed in an apoplast, comprising a step of injecting a buffer into a plant body that expresses the target protein in an apoplast through a stoma, a water pore, or a cross-section of the plant body; and a step of extracting a buffer present inside the plant body through a stoma, a water pore, or a cross-section.