Signal Peptide Linkers for Recombinant Protein Secretion Bottlenecks
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Solution Overview
Problem
Existing methods for increasing enzyme yields in industrial biotechnology face challenges due to the redundancy in signal peptide sequences, making it difficult to predict secretion efficiency, and the cellular machinery is often overwhelmed by high-level protein production, leading to bottlenecks in secretion.
Innovation Solution
Incorporating a signal peptide linker (SP-linker) between the signal peptide and the polypeptide of interest, which facilitates more effective cleavage during maturation, thereby enhancing secretion efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If high-level production of a given protein is achieved through stronger promoters and multi-copy strains, then expression level is improved, but secretion efficiency deteriorates due to bottlenecks in cellular machinery
Solution Approach 1:
The invention segments the signal peptide function by introducing a separate SP-linker element between the signal peptide and the polypeptide of interest. This segmentation allows the signal peptide to focus on targeting while the SP-linker facilitates efficient cleavage and secretion, resolving the bottleneck in cellular machinery overwhelmed by high-level production
Solution Approach 2:
The SP-linker acts as an intermediary element between the signal peptide and the polypeptide of interest. It mediates the transition from signal peptide to mature protein, facilitating more effective cleavage by signal peptidase and improving secretion efficiency without compromising high expression levels
2Ease of manufacture
If signal peptide sequences are selected based on redundancy in amino acid sequence, then selection is simplified, but prediction of secretion efficiency becomes difficult
Solution Approach 1:
The invention segments the signal peptide region into two functional parts: the traditional signal peptide for targeting and the new SP-linker for cleavage facilitation. This segmentation creates predictable functional zones, allowing systematic selection and prediction of secretion efficiency based on the specific properties of each segment
Solution Approach 2:
The SP-linker introduces local quality variation at the cleavage site region, creating a distinct functional zone with specific amino acid properties that enhance cleavage efficiency. This local optimization allows for predictable improvement in secretion efficiency while maintaining overall signal peptide functionality
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 a SP-linker results in up to 230% increased secretion of recombinant proteins, maintaining high expression levels even when combined with different signal peptides, and is suitable for various organisms capable of protein secretion and signal peptide cleavage.
Implementation Method 1
The SP-linker facilitates more effective cleavage of the signal peptide during the maturation process of the polypeptide of interest
Data Source
AI summary
The present invention relates to signal peptides, signal peptide-linkers, fusion polypeptides comprising signal peptide-linker, and polynucleotides encoding the signal peptides, signal peptide-linkers, and fusion polypeptides, and to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing the fusion polypeptides, and methods for increasing secretion of a polypeptide of interest.