YhhS Transporter Variant for Higher O-Phosphoserine Export
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Solution Overview
Problem
Existing methods for producing L-cysteine through O-phosphoserine (OPS) fermentation yield is limited due to inefficient export of OPS from the producing microorganism, necessitating the development of a variant with enhanced OPS exporting activity.
Innovation Solution
A polypeptide variant of the YhhS MFS transporter, specifically with substitutions at positions 241, 246, and 330, and amino acids 88 and 207, is engineered to enhance OPS export activity, accompanied by a polynucleotide encoding this variant and a vector for expression in a microorganism, allowing for high-yield OPS production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the wild-type YhhS transporter is used for OPS export, then the basic export function is maintained, but the export efficiency and productivity are insufficient
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues in the YhhS transporter sequence. Three specific substitutions are made: I241T, D246V, and V330I. These parameter changes at the molecular level alter the transporter's properties to enhance OPS export efficiency while preserving its fundamental transport function and stability.
2Productivity
If multiple amino acid substitutions are introduced to enhance OPS export activity, then the export function is improved, but the protein structure and stability may be affected
Solution Approach 1:
The patent applies local quality by making targeted amino acid substitutions at specific positions (241, 246, and 330) within the polypeptide sequence. Rather than random mutations, the changes are localized to specific regions that likely influence OPS binding or transport. This localized modification approach enhances export activity while minimizing disruption to the overall protein structure and stability.
Data Source
AI summary
An O-phosphoserine (OPS) export protein variant with increased O-phosphoserine exporting activity, and a method for producing O-phosphoserine, cysteine, and cysteine derivatives using the same. Also provided herein are O-phosphoserine-producing microorganisms, a polynucleotide encoding the O-phosphoserine export protein variant, and a vector containing the polynucleotide encoding the O-phosphoserine export protein variant.