Lactobacillus Expression Cassette Using PtlpA for High Protein Yield

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Solution Overview

Problem

The lack of well-characterized genetic tools and insufficient understanding of biochemical pathways in Lactobacillus limit the expression of proteins at high yields, hindering applications in medical and industrial fields.

Innovation Solution

Development of an expression cassette with a promoter operatively linked to a nucleic acid encoding a protein, specifically using the PtlpA promoter from Salmonella typhimurium, which is substantially identical to sequence ID No. 1, and optionally including a ribosome binding site and toxin-antitoxin systems for enhanced expression and plasmid retention.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional promoters are used in Lactobacillus, then protein expression is achieved, but expression yield is insufficient for high-value applications

Engineering Contradiction:

Improveprotein expression yieldVSAvoidexpression system reliability

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent changes the key parameter of promoter sequence identity by providing a promoter with at least 80% identity to the PtlpA promoter sequence, thereby achieving high-level protein expression in Lactobacillus species while maintaining the ability to function across different strains and species

Inventive Principle:

Principle #35Parameter changes

2Adaptability or versatility

If genetic tools are expanded for Lactobacillus engineering, then application capabilities are improved, but tool complexity and characterization requirements increase

Engineering Contradiction:

Improvegenetic tool applicabilityVSAvoidgenetic tool complexity

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent creates a universal promoter tool that functions across multiple Lactobacillus species and strains, enabling broad applicability for protein expression, metabolic engineering, and synthetic biology applications without requiring species-specific optimization

Inventive Principle:

Principle #6Universality (Multi-functionality)

Solution Approach 2:

The promoter sequence is provided in discrete, characterizable units with defined identity thresholds (80%, 90%, 95%, 99% identity), allowing researchers to select appropriate versions based on specific application requirements while maintaining systematic organization

Inventive Principle:

Principle #1Segmentation

3Adaptability or versatility

If plasmids are used for protein expression in Lactobacillus, then functional versatility is achieved, but plasmid retention and stability over generations are insufficient

Engineering Contradiction:

Improveexpression system versatilityVSAvoidplasmid retention duration

Core Design Contradiction:

Adaptability or versatilityVSDuration of action of stationary object

Solution Approach 1:

The patent introduces a toxin-antitoxin system as an intermediary mechanism that mediates plasmid retention by creating a selective pressure that maintains plasmid stability in Lactobacillus populations over multiple generations, thereby resolving the instability issue of expression plasmids

Inventive Principle:

Principle #24Intermediary (Mediator)

Data Source

PatentUS20260071226A1Novel Genetic Tools

Publication Date: 2026.03.12 LEIBNIZ INSTITUT FUR NEUE MATERIALIEN GMBH

AI summary

An expression cassette for expression in Lactobacillus includes a promoter operatively linked to a nucleic acid encoding a protein to be expressed, the promoter being substantially identical to the PtlpA promoter.