Polypeptide Production via Cysteine Sulfinic Acid Decarboxylase Overexpression
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Solution Overview
Problem
Current methods for producing recombinant proteins in animal cells are limited by low yield and efficiency, and there is a lack of understanding on how increased β-alanine concentration affects protein production.
Innovation Solution
Using cells that strongly express cysteine sulfinic acid decarboxylase and a taurine transporter to increase β-alanine production, which enhances the yield of desired polypeptides by neutralizing fatigue substances and maintaining cell survival.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If animal cells are used for recombinant protein production, then post-translational modification and folding capabilities are improved, but production yield and efficiency deteriorate
Solution Approach 1:
The invention changes the intracellular chemical environment by increasing β-alanine concentration through overexpression of cysteine sulfinic acid decarboxylase. This parameter change (β-alanine concentration) improves both cell survival and protein production yield, resolving the contradiction between maintaining modification capability and improving productivity.
2Ease of manufacture
If conventional cell culture methods are used, then production process simplicity is maintained, but cell survival ratio and production efficiency deteriorate due to fatigue substance accumulation
Solution Approach 1:
The invention enables cells to self-neutralize fatigue substances (lactate and ammonia) through endogenous β-alanine production. The overexpressed cysteine sulfinic acid decarboxylase catalyzes cysteine conversion to β-alanine, which then neutralizes acidic and basic fatigue substances, improving cell survival without adding external agents or complicating the process.
3Reliability
If β-alanine is added to culture medium, then antioxidant action and pH buffering are improved, but intracellular β-alanine concentration increase is insufficient
Solution Approach 1:
The invention performs preliminary action by pre-installing the biosynthetic capability within cells through cysteine sulfinic acid decarboxylase overexpression. This enables cells to autonomously produce β-alanine in advance, ensuring sufficient intracellular concentration for neutralizing fatigue substances, rather than relying on external addition that fails to achieve adequate intracellular levels.
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 significantly increases the yield of recombinant proteins by improving β-alanine concentration within cells, leading to higher antibody production and maintaining cell survival ratios, even in the absence of added β-alanine.
Implementation Method 1
cysteine sulfinic acid decarboxylase (CSAD) (EC 4.1.1.29)...synthesizing β-alanine from aspartic acid
Implementation Method 2
β-alanine, by itself, has pH-buffering action and antioxidant action
Implementation Method 3
β-alanine, by itself, has pH-buffering action and antioxidant action
Data Source
AI summary
The present invention provides a method capable of producing a natural or recombinant protein in high yield. The present invention relates to a method of producing a polypeptide, comprising culturing a cell which strongly expresses cysteine sulfinic acid decarboxylase and has a transferred DNA encoding a desired polypeptide and thereby allowing the cell to produce the polypeptide. Hamster cysteine sulfinic acid decarboxylase, a DNA encoding the same, a recombinant vector and a transformed cell are also provided.