Symmetrical Bola Sophorosides Production via Strain Engineering
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Solution Overview
Problem
Current methods for producing bola sophorosides using Starmerella bombicola yield significant amounts of contaminating sophorolipids and non-symmetrical bolaform sophorolipids, with a need for improved strain modifications to enhance production of fully symmetrical, acetylated bola sophorosides.
Innovation Solution
A mutated Starmerella bombicola strain with dysfunctional CYP52M1 cytochrome P450 monooxygenase and FAO1 fatty alcohol oxidase is used, which is fed with fatty alcohols to produce fully symmetrical bola sophorosides free from contaminating sophorolipids, achieving exclusive terminal linkage of sophorose units on the hydrophobic linker.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If a S. bombicola strain deficient in acetyltransferase (at), lactonase esterase (sble) and alcohol oxidase (fao1) is used to produce bola-sophorosides, then production of bola-sophorosides is achieved, but substantial amounts of bolaform sophorolipids are still produced as contaminants
Solution Approach 1:
The patent applies the 'Taking out' principle by selectively removing specific enzymatic functions (at, sble, fao1) from the yeast strain to eliminate the production pathway for unwanted sophorolipid contaminants, while preserving the pathway for desired bola-sophoroside production. This targeted gene deletion strategy extracts the harmful metabolic pathway while maintaining the useful one.
Solution Approach 2:
The patent changes the metabolic parameters of the yeast strain by modifying gene expression levels and enzymatic activities. Specifically, it alters the functional state of key enzymes (acetyltransferase, lactonase esterase, alcohol oxidase) to shift the metabolic flux from sophorolipid production to bola-sophoroside production, achieving high purity products through parameter optimization.
2Productivity
If bola sophorosides are produced by the strain described by Van Renterghem, then production occurs, but the molecules are non-acetylated and non-symmetrical with mixed linkage types
Solution Approach 1:
The patent applies 'Local quality' by introducing site-specific modifications to achieve uniform terminal linkage of sophorose units. The engineered strain produces molecules with consistent structural characteristics (terminal linkages only) at specific positions, creating symmetrical bola-sophorosides with defined acetylation patterns rather than mixed structural isomers.
Solution Approach 2:
The patent inverts the metabolic pathway by blocking the conventional sophorolipid synthesis route and redirecting flux toward bola-sophoroside formation. By eliminating the ester bond formation pathway and promoting glycosidic bond formation, the strain produces acetylated symmetrical structures through an alternative metabolic route.
3Productivity
If alkyl sophorosides production is increased by deleting fao1 gene, then production increases, but acidic and lactonic sophorolipids are still produced which is undesired
Solution Approach 1:
The patent converts the potentially harmful by-product formation into a benefit by completely eliminating the pathways that generate acidic and lactonic sophorolipids. The triple knockout strategy transforms the metabolic flux that would otherwise produce contaminants into productive formation of pure alkyl sophorosides, turning a harmful side reaction into a beneficial selective pathway.
Data Source
AI summary
The present invention relates to the field of microbial production of novel biosurfactants. More specifically, the present invention discloses the usage of a fungal strain such as the yeast Starmerella bombicola having a dysfunctional CYP52M1 cytochrome P450 monooxygenase and a dysfunctional FAO1 fatty alcohol oxidase for producing high amounts of so-called “symmetrical bolaform sophorosides” where both sophorose moieties are attached through a terminal glycosidic linkage to the hydrophobic linker. In addition, the present invention further discloses that the latter yeast can also be used to produce alkyl sophorosides and symmetrical bolaform glucosides.