4-Hydroxyproline Fermentation from Plant Cells
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Solution Overview
Problem
Current methods for obtaining 4-hydroxyproline from animal sources are contaminated with bioactive peptides, posing risks of diseases like bovine spongiform encephalopathy and are not suitable for biological agriculture, necessitating a contamination-free alternative source.
Innovation Solution
A method involving the fermentation of plant cell cultures, specifically BY2 tobacco cells, to obtain 4-hydroxy-L-proline compositions that are free from biological contaminants, using a process that includes fermentation, separation, lysis of cellular walls, and enzymatic or chemical digestion to produce peptides and amino acids, followed by purification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If animal source proteins are used to obtain 4-hydroxyproline, then the aminoacid can be produced, but the composition is contaminated with bioactive peptides and dangerous pathogens like prions
Solution Approach 1:
The patent extracts and removes harmful components (sugar chains, bioactive peptides, prions) from the protein matrix through selective hydrolysis and purification processes, isolating 4-hydroxyproline in a contaminant-free form from plant cell wall proteins
Solution Approach 2:
The patent uses plant cell wall proteins as an intermediary source instead of direct animal proteins, introducing a safe intermediate host that produces 4-hydroxyproline without containing harmful pathogens like prions
2Productivity
If acid hydrolysis is performed at high temperature to accelerate the process, then 4-hydroxyproline is released, but racemization may occur
Solution Approach 1:
The patent optimizes hydrolysis parameters by performing the process at controlled temperatures (70-100°C) and controlled pH levels, using enzyme catalysis instead of extreme acid conditions, thereby maintaining high productivity while preventing racemization and preserving aminoacid configuration integrity
3Manufacturing precision
If multiple purification steps are performed to remove contaminants, then 4-hydroxyproline purity increases, but the process complexity and time increase
Solution Approach 1:
The patent segments the purification process into distinct functional stages: cell wall isolation, selective hydrolysis, filtration, and chromatographic purification, allowing each step to target specific contaminants systematically while maintaining overall process efficiency
Solution Approach 2:
The patent replaces complex mechanical separation methods with biochemical selectivity, using enzyme-specific hydrolysis and affinity chromatography to achieve high purity 4-hydroxyproline with fewer and simpler purification 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
The method provides 4-hydroxy-L-proline compositions that are free from biological contaminants, capable of inducing plant defense genes and enhancing stress tolerance in crops, offering a safe and effective alternative for agricultural use.
Implementation Method 1
The method involves the fermentation of plant cell cultures, specifically BY2 tobacco cells, to obtain 4-hydroxy-L-proline compositions
Implementation Method 2
separation, lysis of cellular walls, and enzymatic or chemical digestion to produce peptides and amino acids
Implementation Method 3
enzymatic or chemical digestion to produce peptides and amino acids
Implementation Method 4
The methods for obtaining oligopeptides rich in hydroxyproline envisage that the protein source of animal origin be initially ground and preserved in low-temperature environments until the moment of treatment, which comprises the following phases: Acid hydrolysis
Implementation Method 5
capable of inducing plant defense genes and enhancing stress tolerance in crops
Implementation Method 6
enhancing stress tolerance in crops
Data Source
AI summary
The present invention relates to a method for the preparation of 4-hyroxy-L-proline or compositions with a high content of this aminoacid and the use thereof as agrochemical in the agronomical field and/or as a synthesis intermediate of chemical compounds.