Thioesterase Variant Engineering for Lipid Productivity
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Solution Overview
Problem
Current methods for enhancing lipid productivity in animals and plants are limited in efficiency, as existing enzymes such as acetyl-CoA carboxylase, yeast sn-2 acyltransferase, and diacylglycerol acyltransferase do not significantly improve lipid content beyond a certain threshold.
Innovation Solution
A thioesterase variant with a modified amino acid sequence, specifically altering the 209th amino acid from tryptophan to threonine, glutamic acid, or alanine, and allowing for deletions, substitutions, or additions in the 112nd to 414th amino acid region, is introduced into hosts to enhance lipid production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If existing enzymes (acetyl-CoA carboxylase, yeast sn-2 acyltransferase, diacylglycerol acyltransferase) are introduced to enhance lipid content, then lipid content increases to a certain threshold, but productivity cannot be significantly improved beyond that threshold
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the thioesterase enzyme at specific positions (particularly position 209). This molecular-level parameter change results in enhanced enzymatic activity and substrate specificity, enabling the enzyme to process fatty acid chains more efficiently and release them from ACP, thereby significantly improving lipid productivity beyond what wild-type enzymes can achieve.
Solution Approach 2:
The patent creates modified copies of the wild-type thioesterase enzyme by introducing specific amino acid substitutions. These engineered enzyme variants retain the core function of the original enzyme but possess improved catalytic properties. The modified enzymes are then introduced into host organisms to achieve enhanced lipid production, effectively using improved copies to overcome the limitations of the original enzyme system.
2Productivity
If thioesterase amino acid sequence is modified to enhance lipid production, then lipid productivity increases, but enzyme structure complexity increases
Solution Approach 1:
The patent applies local quality by making targeted amino acid substitutions at specific positions (particularly position 209) within the thioesterase enzyme structure, rather than modifying the entire enzyme. This localized modification approach allows the enzyme to maintain its overall stable structure and folding while introducing specific functional improvements at critical catalytic sites, thus achieving enhanced productivity without proportionally increasing overall structural complexity.
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 modified thioesterase variant significantly increases lipid productivity compared to wild-type thioesterase, allowing for higher yields of fatty acids and lipids, particularly those with 12 to 16 carbon atoms, suitable for industrial applications such as surfactants and cosmetics.
Implementation Method 1
a protein comprising an amino acid sequence of the 112nd to 414th amino acids set forth in SEQ ID NO: 1 in which an amino acid corresponding to the 209th amino acid set forth in SEQ ID NO: 1 is substituted from tryptophan to threonine, glutamic acid or alanine, and having thioesterase activity
Data Source
AI summary
A method of producing a lipid, comprising steps ofintroducing a gene that encodes the following protein (a) or (b) into a host, and thereby obtaining a transformant, andcollecting a lipid from the transformant:(a) A protein comprising an amino acid sequence of the 112nd to 414th amino acids set forth in SEQ ID NO: 1 in which an amino acid corresponding to the 209th amino acid set forth in SEQ ID NO: 1 is substituted from tryptophan to threonine, glutamic acid or alanine, and having thioesterase activity, and(b) A protein comprising an amino acid sequence of the protein (a) in which one to several amino acids other than the amino acid corresponding to the 209th amino acid set forth in SEQ ID NO: 1 are deleted, substituted, inserted or added, and having thioesterase activity.