Lactobacillus Surface Expression Vector for Stable Protein Co-Expression

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Solution Overview

Problem

Current methods for expressing foreign proteins on microbial surfaces, particularly using lactic acid bacteria, face challenges such as insufficient expression levels and unsustainable protein production due to the limitations of inducible promoters, and the need for efficient co-expression of multiple proteins for vaccine development and antigen presentation.

Innovation Solution

A surface expression vector utilizing a poly-gamma-glutamate synthetase complex from Bacillus subtilis and promoters from Lactobacillus casei, which co-expresses two target proteins on the microbial surface, minimizing cellular transformation steps and enabling stable, efficient expression of foreign proteins.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If inducible promoters are used for protein expression in lactic acid bacteria, then protein expression can be controlled, but expression levels are insufficient and protein production is unsustainable

Engineering Contradiction:

Improvesustainability of protein productionVSAvoidexpression level of target protein

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent changes the promoter parameters from inducible promoters (with limited expression levels) to strong constitutive promoters (lac, recA, or gap) that continuously drive high-level expression of target proteins, thereby resolving the contradiction between sustainability and expression level

Inventive Principle:

Principle #35Parameter changes

2Productivity

If multiple transformation steps are used to co-express multiple proteins, then each protein can be expressed individually, but the process complexity increases and efficiency decreases

Engineering Contradiction:

Improveco-expression efficiency of multiple proteinsVSAvoidnumber of transformation steps

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent merges multiple separate transformation steps into a single transformation step by designing a vector system where multiple target genes are linked to different constitutive promoters within the same plasmid, allowing simultaneous expression of multiple proteins in one transformation event

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The patent creates a universal vector backbone with multiple constitutive promoter options that can accommodate various target genes, enabling the same vector system to co-express different combinations of proteins through a single transformation protocol

Inventive Principle:

Principle #6Universality (Multi-functionality)

Data Source

PatentUS12188027B2Surface expression vector using two kinds of promoters derived from <i>Lactobacillus casei </i>for concurrently expressing two target proteins and method for expressing proteins on microbial surface by using same

Publication Date: 2025.01.07 BIOLEADERS CORP

AI summary

Provided is a vector capable of co-expressing two different target proteins on the microbial surface using two promoters derived from Lactobacillus, and a method of expressing target proteins on the microbial surface using the vector. The vector containing foreign genes inserted therein is transformed into a microorganism, and allows different foreign proteins to be stably expressed on the surface of the microorganism. Furthermore, provided is a surface expression vector containing the gene pgsA encoding a poly-gamma-glutamate synthetase complex, and a method of expressing a target protein on the microbial surface using the vector. The vector containing foreign genes inserted therein is transformed into a microorganism, and allows the foreign proteins to be stably expressed on the surface of the microorganism.