Microbial Sphingoid Base Production via Toxin Selection
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Solution Overview
Problem
Current methods for producing sphingosine and sphinganine are costly due to the need for extensive protection chemistry and result in racemic mixtures with only 25% of the naturally occurring stereochemical configuration, making them unsuitable for food and cosmetic applications, while natural sources are expensive and potentially unsafe.
Innovation Solution
Development of a microbial strain, specifically a yeast strain like Pichia ciferrii, that produces at least 0.1 mg per g biomass dry weight of sphingoid bases such as sphingosine and sphinganine, using toxin resistance selection and genetic engineering to enhance production levels and alter metabolic pathways.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If chemical synthesis is used to produce sphingosine and sphinganine, then production cost is reduced through standardized processes, but the product becomes a racemic mixture with only 25% naturally occurring stereochemical configuration
Solution Approach 1:
The patent changes the fundamental production parameter from chemical synthesis to microbial fermentation, transforming the production system rather than optimizing the existing chemical process. This allows obtaining naturally occurring stereochemical configuration through biological systems while maintaining production efficiency.
Solution Approach 2:
The patent uses microbial strains that naturally produce sphingoid bases with the correct stereochemical configuration, copying the natural biosynthetic pathway. This avoids the need for complex chiral resolution or asymmetric synthesis while achieving high stereopurity.
2Manufacturing precision
If extensive protection chemistry is applied during chemical synthesis, then manufacturing precision is improved, but device complexity and production cost increase
Solution Approach 1:
The patent replaces complex chemical synthesis mechanisms with biological fermentation mechanisms. The microbial strain naturally performs the complex chemical transformations needed to produce sphingoid bases with correct stereochemistry, eliminating the need for multiple protection and deprotection steps.
Solution Approach 2:
The microbial strain self-regulates the biosynthetic pathway, naturally producing sphingoid bases with the correct stereochemical configuration without requiring external intervention for chiral control. The biological system inherently performs the function that would otherwise require complex chemical processing.
3Manufacturing precision
If pure sphingoid bases are isolated from natural sources, then manufacturing precision is improved, but productivity is reduced due to limited availability
Solution Approach 1:
The patent uses a microbial strain that can be cultured in large volumes to produce sphingoid bases, making the production system scalable. The same biological system that ensures high purity through natural stereochemistry can also be expanded to meet large-scale production demands.
Solution Approach 2:
The patent changes the production scale parameter from limited natural extraction to large-scale microbial fermentation. By using microorganisms that can be cultivated in fermenters, the system achieves both high purity and high productivity simultaneously.
4Productivity
If heterogeneous sphingolipid preparations are extracted from animal sources, then productivity is improved through abundant natural sources, but reliability worsens due to compositional heterogeneity and potential pathogenic agents
Solution Approach 1:
The patent extracts the desirable production capability from natural sources and transfers it to a controlled microbial system. By using genetically engineered microorganisms, the system achieves consistent, pure production without the contaminants and heterogeneity associated with animal source extraction.
Solution Approach 2:
The patent changes the source organism parameter from animal sources to microbial sources. This fundamental parameter change enables controlled, sterile production with consistent composition while maintaining high availability through scalable fermentation processes.
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 microbial strain achieves high productivity of sphingoid bases, allowing for efficient and cost-effective production suitable for commercial use in food, pharmaceutical, and cosmetic industries, overcoming the limitations of chemical synthesis and natural source extraction.
Implementation Method 1
The yeast Pichia ciferrii was shown to produce rather high levels of sphingoid bases and derivatives thereof
Data Source
AI summary
The present invention provides microbial strains, in particular yeast strains, that produce at least 0.1 mg per g biomass dry weight of a sphingoid base. The present invention further provides a method to obtain sphingoid base-producing microbial strains comprising incubating a population of microbial cells in the presence of a suitable concentration of a toxin, selecting cells that are resistant against said toxin, and isolating cells out of the toxin-resistant cell population that produce at least 0.1 mg per g biomass dry weight of the sphingoid base of Formula I. Optionally, the method further comprises subjecting a population of toxin-resistant microbial cells that produce at least 0.1 mg per g biomass dry weight of the sphingoid base of Formula I to DNA-mediated transformation with a polynucleotide encoding an enzyme of the sphingolipid metabolic pathway. The present invention further provides a polypeptide having dihydroceramide desaturase activity obtainable form Pichia ciferrii.