Microbial Production of (3R)-Hydroxybutyl (3R)-Hydroxybutyrate
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for producing (3R)-hydroxybutyl (3R)-hydroxybutyrate face challenges such as low yields, impurity issues, and impracticality on a large scale due to complex chemical synthesis processes, making it difficult to achieve the necessary therapeutic blood plasma levels for clinical benefits.
Innovation Solution
Development of non-naturally occurring microbial organisms with engineered pathways that include exogenous nucleic acids encoding enzymes for (3R)-hydroxybutyl (3R)-hydroxybutyrate production, allowing for biosynthesis of the compound in a more efficient and scalable manner.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If classical chemical synthesis from poly[(3R)-hydroxybutyric acid] is used, then pure product is obtained, but the process involves six chemical steps making it complex and impractical on large scale
Solution Approach 1:
The patent replaces complex chemical synthesis mechanisms with biological enzymatic mechanisms. Engineered microorganisms express specific enzymes (horseradish peroxidase, lactoperoxidase, or microperoxidase) that catalyze the formation of (3R)-hydroxybutyl (3R)-hydroxybutyrate from (3R)-hydroxybutyrate, eliminating the need for six separate chemical synthesis steps while maintaining product purity.
Solution Approach 2:
The patent changes the fundamental parameter of synthesis approach from chemical to biological. By using peroxidase enzymes under physiological conditions (aqueous environment, moderate temperature, pH 6-8), the process achieves the same synthetic outcome with dramatically reduced complexity and improved scalability.
2Quantity of substance
If existing synthesis methods are used, then some product is produced, but yields are low making it difficult to achieve therapeutic blood plasma levels
Solution Approach 1:
The engineered microorganisms serve as self-contained production systems. They take simple substrates (such as (3R)-hydroxybutyrate or its precursors) and convert them to the desired product through their metabolic pathways, which include engineered enzymes like peroxidases. This self-service capability enables high-yield production without requiring complex external intervention.
Solution Approach 2:
The patent employs peroxidase enzymes that can function with multiple substrates and under various conditions. These enzymes are capable of catalyzing the formation of (3R)-hydroxybutyl (3R)-hydroxybutyrate from different precursor combinations, providing a universal and flexible production platform that achieves high yields and productivity.
3Reliability
If direct administration of ketone bodies is performed, then therapeutic benefits are achieved, but significant acidosis occurs following rapid absorption
Solution Approach 1:
The patent uses (3R)-hydroxybutyl (3R)-hydroxybutyrate as an intermediary compound. This ester prodrug is administered instead of direct ketone bodies, and it is gradually hydrolyzed in the body to release (3R)-hydroxybutyrate. This intermediary approach allows controlled release, achieving therapeutic ketone body levels while avoiding the rapid absorption-induced acidosis associated with direct administration.
4Quantity of substance
If sodium salt of ketone bodies is administered, then therapeutic levels are reached, but dangerous sodium overload occurs
Solution Approach 1:
The (3R)-hydroxybutyl (3R)-hydroxybutyrate ester serves as a sodium-free intermediary. When administered, it provides ketone bodies without requiring sodium counterions. The ester is hydrolyzed endogenously to release free (3R)-hydroxybutyrate, achieving therapeutic ketone body levels without the sodium overload problem inherent in administering sodium salts.
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 organisms enable the production of purified (3R)-hydroxybutyl (3R)-hydroxybutyrate, achieving therapeutic levels in the blood plasma, thereby addressing the limitations of existing synthesis methods and providing clinical benefits like enhanced energy production and treatment of various conditions.
Implementation Method 1
the organism includes at least one exogenous nucleic acid encoding a (3R)-hydroxybutyl (3R)-hydroxybutyrate pathway enzyme disclosed herein
Implementation Method 2
methods and processes for producing (3R)-hydroxybutyl (3R)-hydroxybutyrate using the microbial organisms
Data Source
AI summary
Provided herein are non-naturally occurring microbial organisms having a pathway for production of (3R)-hydroxybutyl (3R)-hydroxybutyrate, wherein the organism can further include a (R)-1,3-butanediol pathway, a (3R)-hydroxybutyrate pathway, a (3R)-hydroxybutyryl-CoA pathway, an acetoacetate pathway, an acetoacetyl-CoA pathway, a (3R)-hydroxybutyl-ACP pathway, or an acetoacetyl-ACP pathway. Additionally provided are methods and processes for producing and isolating (3R)-hydroxybutyl (3R)-hydroxybutyrate using the microbial organisms, and various compositions having the (3R)-hydroxybutyl (3R)-hydroxybutyrate. Still further provided are methods of treating or preventing a disease, disorder or condition using the (3R)-hydroxybutyl (3R)-hydroxybutyrate produced by the microbial organisms of the invention.