Bacillus Culture Broth with Inhibition-Resistant Prephenate Dehydratase
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Solution Overview
Problem
Current plant growth-promoting bacteria, such as Bacillus amyloliquefaciens FZB42, face limitations in growth promotion due to feedback inhibition by phenylalanine, which reduces their effectiveness in enhancing plant growth.
Innovation Solution
A culture broth of Bacillus bacteria with prephenate dehydratase resistant to feedback inhibition by phenylalanine is applied to plants, specifically strains like Bacillus amyloliquefaciens or Bacillus velezensis with mutations at positions S213, N229, or F247, to enhance plant growth by reducing feedback inhibition and increasing phenylalanine production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If prephenate dehydratase is used for phenylalanine production, then phenylalanine production increases, but feedback inhibition by phenylalanine reduces enzyme activity and limits further production
Solution Approach 1:
The patent applies parameter changes by introducing specific mutations (S213L, N229Y, F247Y) in the prephenate dehydratase enzyme to alter its biochemical properties. These mutations change the enzyme's sensitivity to phenylalanine feedback inhibition, allowing it to maintain high catalytic activity even when phenylalanine accumulates to high concentrations, thereby resolving the contradiction between increasing production and maintaining enzyme stability
Solution Approach 2:
The patent converts the harmful feedback inhibition effect into a beneficial feature by creating mutant enzymes that are resistant to phenylalanine inhibition. The very substance (phenylalanine) that normally inhibits the enzyme becomes a marker for the enzyme's resistance capability, allowing the system to tolerate and even thrive under conditions of high phenylalanine production, thus turning the harmful feedback mechanism into a beneficial trait for industrial production
2Productivity
If conventional Bacillus amyloliquefaciens FZB42 is applied to plants, then some plant growth promotion occurs, but effectiveness is limited due to feedback inhibition
Solution Approach 1:
The patent introduces specific amino acid mutations (S213L, N229Y, F247Y) in the prephenate dehydratase enzyme of Bacillus amyloliquefaciens and Bacillus velezensis strains. These parameter changes in enzyme structure fundamentally alter the bacteria's metabolic capabilities, enabling them to produce phenylalanine without feedback inhibition, thereby significantly enhancing their ability to promote plant growth compared to conventional strains
Solution Approach 2:
The modified Bacillus strains perform self-service by autonomously producing phenylalanine and other plant growth-promoting substances through their engineered metabolic pathways. The bacteria's own cellular machinery, equipped with mutation-resistant prephenate dehydratase, continuously synthesizes these beneficial compounds without being self-inhibited, allowing them to independently and consistently promote plant growth in agricultural applications
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 application of the culture broth with inhibition-resistant prephenate dehydratase significantly increases lateral root formation and plant growth, as demonstrated by improved growth metrics in Arabidopsis thaliana, indicating enhanced plant growth promotion.
Implementation Method 1
Prephenate dehydratase (PD) is one of the phenylalanine biosynthesis enzymes
Implementation Method 2
Application of a culture broth of a Bacillus bacterium having prephenate dehydratase (PD) resistant to feedback inhibition by phenylalanine to a plant has been found to improve growth of the plant
Data Source
AI summary
An agricultural or horticultural composition is provided. A composition containing a culture broth of a Bacillus bacterium having prephenate dehydratase (PD) resistant to feedback inhibition by phenylalanine.