Genetically Engineered Yeast for High NADPH Lactate Production
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Solution Overview
Problem
Current methods for producing lactate face challenges in achieving high yields due to homeostasis issues in microorganisms, necessitating a strain capable of efficient lactate production that can overcome these limitations.
Innovation Solution
A genetically engineered yeast cell with increased NADPH production, achieved through genetic modifications such as overexpressing specific proteins like ADE3, SHM2, MTD1, UTR1, YEF1, POS5, and ZWF1, and introducing exogenous genes like GDP1 and SthA, along with disruptions in genes involved in competing pathways, to enhance lactate dehydrogenase activity and NADPH levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional fermentation methods are used to produce lactate, then the process is simple and uses standard microorganisms, but the lactate yield is limited due to homeostasis constraints
Solution Approach 1:
The patent changes the biochemical parameters of the yeast cell by introducing exogenous genes (G6PDH, ZWF1, F6P) to enhance the pentose phosphate pathway, thereby increasing NADPH production capacity. This parameter change enables the microorganism to overcome homeostatic limitations and achieve higher lactate yields without changing the fundamental fermentation process
Solution Approach 2:
The patent introduces an intermediary metabolic pathway (pentose phosphate pathway) as a mediator between glucose consumption and lactate production. By enhancing this intermediate pathway through exogenous gene introduction, the system can generate sufficient NADPH to support high-rate lactate production while maintaining cellular homeostasis
2Productivity
If exogenous genes are introduced to enhance NADPH production, then lactate production capability increases, but the genetic modification complexity increases
Solution Approach 1:
The patent segments the lactate production enhancement into distinct functional modules: (1) exogenous genes for NADPH generation (G6PDH, ZWF1, F6P), (2) lactate dehydrogenase gene (ldh) for lactate synthesis, and (3) regulatory elements for coordinated expression. This modular segmentation makes the complex genetic modification more manageable and implementable
3Productivity
If homeostasis is maintained in standard microorganisms, then cell viability is preserved, but lactate production efficiency is limited
Solution Approach 1:
The patent performs preliminary action by pre-engineering the yeast strain with exogenous genes before the fermentation process. The enhanced pentose phosphate pathway is established in advance, ensuring that NADPH is readily available when lactate production is initiated, thereby avoiding homeostatic conflicts during actual production
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 genetically engineered yeast cell demonstrates increased lactate production capabilities, with enhanced glucose consumption and lactate yields compared to wild-type strains, under microaerobic conditions, effectively addressing the homeostasis challenges in microorganism-based lactate production.
Implementation Method 1
The biotechnological fermentation process is used to manufacture lactate from a reproducible carbon hydrate, such as starch, sucrose, maltose, glucose, fructose, or xylose, as a substrate
Implementation Method 2
enhance lactate dehydrogenase activity and NADPH levels
Data Source
AI summary
Provided is a genetically engineered yeast cell having increased NADPH production, a method of increasing a NADPH level in a yeast cell, a method of preparing the genetically engineered yeast cell, and a method of producing lactate using the genetically engineered yeast cell.