Schizosaccharomyces pombe Transformant for Malic Acid Production
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Solution Overview
Problem
Current transformants for producing malic acid through microbial fermentation have malic acid production rates of less than 1.0 g/(L·h), necessitating the development of a microorganism with higher producibility.
Innovation Solution
A Schizosaccharomyces pombe transformant is created by incorporating foreign genes such as phosphoenolpyruvate carboxykinase (PCK) and pyruvate carboxylase (PYC) genes, while deleting or deactivating pyruvate decarboxylase (PDC) and malic enzyme (MAE) genes, to enhance malic acid production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If gene recombination is used to improve malic acid producibility, then production rate increases, but existing transformants still achieve less than 1.0 g/(L·h)
Solution Approach 1:
The invention changes key metabolic parameters by introducing foreign genes (PCK, PYC, MDH) and deleting native genes (PDC, MAE) to fundamentally alter the metabolic pathway, achieving a production rate of 2.3 g/(L·h) that exceeds the 1.0 g/(L·h) threshold
Solution Approach 2:
The transformant combines multiple genetic modifications (PCK gene introduction, PYC gene introduction, PDC gene deletion, MAE gene deletion) to create a composite genetic system that synergistically enhances malic acid production beyond what single modifications could achieve
2Productivity
If multiple gene modifications are introduced to enhance production, then productivity improves, but the genetic engineering process becomes more complex
Solution Approach 1:
The genetic modification process is segmented into distinct functional modules: PCK gene introduction for phosphoenolpyruvate carboxykinase activity, PYC gene introduction for pyruvate carboxylase activity, PDC gene deletion to block ethanol pathway, and MAE gene deletion to prevent malate degradation. Each module addresses a specific metabolic step, making the complex engineering process more manageable and systematic
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 transformant achieves a higher malic acid production rate and improved productivity through microbial fermentation, exceeding previous rates and simplifying the manufacturing process.
Implementation Method 1
a phosphoenolpyruvate carboxykinase gene which encodes a polypeptide having phosphoenolpyruvate carboxykinase activity
Implementation Method 2
a pyruvate carboxylase gene which encodes a polypeptide having pyruvate carboxylase activity
Implementation Method 3
a method for manufacturing a C4 dicarboxylic acid in which the transformant is cultured in a culture solution and a C4 dicarboxylic acid is obtained from the culture solution
Data Source
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AI summary
Provided is a transformant which uses Schizosaccharomyces pombe as a host, into which one or more foreign genes selected from the group consisting of a phosphoenolpyruvate carboxykinase gene and a pyruvate carboxylase gene are incorporated, and in which pdc2 genes of the Schizosaccharomyces pombe host have undergone deletion or deactivation, and a method for manufacturing a dicarboxylic acid having 4 carbon atoms by using the transformant.