Artificial Secretion Peptides for Lactobacillus Protein Production
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Solution Overview
Problem
Existing therapeutic protein production technologies rely on naturally-occurring secretion signal peptides, which are unpredictable and inefficient, and their performance is not transferable between different proteins or bacterial strains, leading to inefficient export and misfolding of proteins.
Innovation Solution
Development of artificial secretion signal peptides with specific amino acid sequences, such as those identified by SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, and SEQ ID NO:7, that are at least 90% identical and capable of directing the secretion of functional heterologous proteins from Lactobacillus cells, including therapeutic proteins like antibodies and cytokines.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If naturally-occurring secretion signal peptides are used, then protein secretion is achieved, but the secretion efficiency is unpredictable and low
Solution Approach 1:
The patent modifies the amino acid sequence parameters of natural signal peptides to create artificial variants with optimized properties. Specifically, the invention identifies and modifies key residues in the signal peptide sequence (such as positions 1-15 in the N-terminal region) to enhance secretion efficiency while maintaining reliability across different proteins and bacterial strains.
Solution Approach 2:
The artificial signal peptides developed in this patent are designed to be universal and transferable across different therapeutic proteins and bacterial strains. The patent demonstrates that a single artificial signal peptide sequence can effectively direct secretion of multiple different heterologous proteins (including antibodies, enzymes, and cytokines) in various bacterial hosts, eliminating the need for protein-specific optimization.
2Productivity
If naturally-occurring secretion signal peptides are used, then some proteins are secreted, but export efficiency and protein folding are compromised
Solution Approach 1:
The patent optimizes specific parameters of the signal peptide sequence, particularly the hydrophobicity of the central region and the charge distribution in the N-terminal region. These parameter changes enhance the interaction with the Sec translocase while ensuring proper protein folding signals are maintained, resulting in both high export efficiency and correct protein folding.
3Productivity
If a signal peptide is optimized for one protein, then that protein is secreted efficiently, but the same signal peptide cannot promote secretion of other proteins
Solution Approach 1:
The artificial signal peptides are designed with a consensus sequence that captures the essential features needed for Sec pathway recognition across different protein substrates. The patent demonstrates that these universal signal peptides can replace protein-specific natural signals, achieving both high secretion efficiency for the target protein and broad applicability to other therapeutic proteins.
4Productivity
If a signal peptide functions in one bacterial strain, then protein secretion is achieved, but it fails to promote secretion in other closely related strains
Solution Approach 1:
The patent modifies the signal peptide sequence parameters to account for variations in Sec translocase recognition across different bacterial strains. By optimizing the hydrophobicity profile and charge distribution in the signal peptide, the invention creates variants that maintain consistent interaction with the Sec machinery across genetically diverse bacterial hosts, including Lactobacillus, E. coli, and other Gram-negative and Gram-positive strains.
Data Source
AI summary
Provided herein, in some embodiments, are artificial secretion peptides capable of directing secretion from Lactobacillus for use, for example, in producing heterologous proteins, including therapeutic proteins.