Isomerase Nucleic Acid Codon Optimization for Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
The production of xylose isomerase at industrial scales is hindered by low expression levels, enzyme stability issues, and high costs due to limitations in fermentation and purification, making it challenging to achieve efficient and cost-effective production of high-value sugars like high fructose syrup.
Innovation Solution
A novel nucleic acid sequence encoding an isomerase enzyme is designed for optimal expression in heterologous hosts, combined with improved vectors and recombinant cells, enabling efficient and scalable production of xylose isomerase with enhanced activity across a wide range of physiological conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional expression systems are used to produce xylose isomerase, then the production process is simple, but the expression level is low leading to low enzyme activity
Solution Approach 1:
The patent applies parameter changes by modifying the nucleic acid sequence parameters including codon optimization for heterologous host expression, adjusting GC content, and optimizing regulatory elements. These parameter modifications enable high-level expression of xylose isomerase in heterologous hosts while maintaining the simplicity of the expression system architecture.
Solution Approach 2:
The patent creates an optimized copy of the xylose isomerase gene with improved nucleic acid sequence features. This copied and enhanced version is then introduced into heterologous hosts, achieving high expression levels without requiring complex expression system modifications.
2Quantity of substance
If xylose isomerase is produced at industrial scale, then the enzyme quantity increases, but stability and purification costs become limiting factors
Solution Approach 1:
The patent performs preliminary action by incorporating stability-enhancing features into the nucleic acid sequence and expression system before industrial-scale production. This includes optimizing the gene sequence for proper folding, adding stability tags, and selecting expression conditions that maintain enzyme stability, thereby enabling large-scale production with maintained reliability.
3Ease of manufacture
If conventional production methods are used, then the process is straightforward, but the production cost is high
Solution Approach 1:
The patent enables self-service by designing a nucleic acid sequence that autonomously achieves high-level expression and maintains enzyme stability without requiring complex external interventions. The optimized sequence includes self-regulatory elements and stability features that allow the system to maintain efficient production independently, reducing manufacturing costs while improving productivity.
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
This approach results in increased expression levels and stability of the isomerase enzyme, leading to cost-effective and efficient production of rare sugars, such as D-allose and D-tagatose, with improved yield and multifunctional isomerization capabilities.
Implementation Method 1
Xylose isomerase, also known as glucose isomerase (GI) or fructose isomerase (FI) is an enzyme in having high industrial application and is amongst the highest tonnage value enzymes. Xylose isomerase has multiple functionality depending on the substrates and catalyzes the inter-conversion of D-xylose to D-xylulose, D-ribose to D-ribulose and D-glucose to D-fructose.
Data Source
AI summary
The present invention provides for a nucleic acid encoding an isomerase and uses of the isomerase for bioconversion of sugar substrates. The invention represents an advancement in the field of enzyme engineering and discloses a modified nucleic acid for achieving optimum expression of a protein having isomerase activity in a heterologous host. The invention also discloses vectors carrying the modified nucleic acid and recombinant host cells carrying the vectors. The invention also discloses the process for producing a recombinant host cell, process for production of the recombinant enzyme and the process for bioconversion of sugars into their respective isomers using the recombinant protein.