Thioesterase Enzymes for Natural Thioester Production
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Solution Overview
Problem
Current methods for producing thioesters, such as methyl butanethioate, methyl thioacetate, and methyl thiopropionate, are not predictable and economically viable, making it difficult to label these compounds as 'natural' for use in food products.
Innovation Solution
Identification of nucleotide sequences encoding thioesterase enzymes, which can be used to produce thioesters efficiently and label them as 'natural' by encoding enzymes with specific amino acid sequences, enabling their use in consumable products and screening for wild-type bacteria capable of forming thioesters.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If thioesters are produced synthetically, then production efficiency is improved, but the ability to label as 'natural' deteriorates
Solution Approach 1:
The patent replaces chemical synthesis methods with biological enzymatic methods. Specifically, thioesterases (enzymes) are used to catalyze the formation of thioesters from thiol and carboxylic acid substrates, substituting the mechanical/chemical synthesis process with a biological system that produces 'natural' labeled compounds while maintaining efficient production
2Ease of manufacture
If thioesters are produced via existing bacterial fermentation, then natural labeling is improved, but production predictability and economic viability deteriorate
Solution Approach 1:
The patent changes key parameters of the fermentation process by introducing specifically identified thioesterase enzymes with known amino acid sequences and nucleotide sequences. This allows control over enzyme activity, substrate specificity, and reaction conditions, making the production process predictable while maintaining natural labeling capability
Solution Approach 2:
The patent identifies and copies specific nucleotide sequences (SEQ ID NOS: 1, 3, 5, and 7) that encode thioesterase enzymes. By replicating these genetic sequences in bacterial systems, the invention creates predictable, reproducible production of thioesters with consistent yields and product ratios
3Ease of manufacture
If existing bacterial fermentation methods are used, then natural production is improved, but yield control and product ratio predictability deteriorate
Solution Approach 1:
The patent employs feedback control by identifying specific thioesterase enzyme sequences and using them to monitor and control the fermentation process. The known enzyme sequences allow for measurement of enzyme activity and adjustment of fermentation conditions to achieve desired yield and product ratio outcomes
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 use of nucleotide sequences encoding thioesterase enzymes allows for the efficient production of thioesters, enhancing the predictability and economic viability of their production, enabling their use in food products while maintaining a 'natural' label.
Implementation Method 1
nucleotide sequences encoding thioesterase enzymes, methods of their production, and their use in methods of forming thioesters
Data Source
AI summary
Disclosed are nucleotide sequences encoding thioesterase enzymes, methods for their production, their use in methods to form thioesters, and their use in methods of screening for other wild type bacteria capable of producing said thioesterase enzymes. Also disclosed are compositions comprising thioesters produced by the methods disclosed herein.