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Genetic engineering of Lactobacillus diolivorans
Stefan Pflügl1, Hans Marx, Diethard Mattanovich
1School of Bioengineering, FH Campus Wien - University of Applied Sciences, Vienna, Austria.
FEMS Microbiology Letters
|May 4, 2013
Summary
We created genetic tools for Lactobacillus diolivorans to produce 1,3-propanediol. This enables enhanced production by overcoming transformation barriers and overexpressing key enzymes.
Area of Science:
- Microbiology
- Synthetic Biology
- Metabolic Engineering
Background:
- Lactobacillus diolivorans is a key organism for producing 1,3-propanediol from glycerol.
- Genetic manipulation tools are essential for optimizing its production capabilities.
Purpose of the Study:
- To develop a genetic toolbox for Lactobacillus diolivorans.
- To enable efficient homologous and heterologous protein expression.
- To enhance 1,3-propanediol production.
Main Methods:
- Developed a novel expression plasmid (pSHM) based on L. diolivorans replication origin.
- Utilized the native glyceraldehyde-3-phosphate dehydrogenase promoter for gene expression.
- Demonstrated functional gene expression via green fluorescent protein and enzyme overexpression.
Main Results:
- Identified absence of native plasmid and DNA methylation as crucial for transformation.
- Constructed pSHM plasmid for robust protein expression.
- Achieved 40-60 fold increase in fluorescence with GFP expression.
- Improved 1,3-propanediol production by 20% through enzyme overexpression.
Conclusions:
- Established a reliable genetic manipulation system for Lactobacillus diolivorans.
- The developed toolbox facilitates metabolic engineering for enhanced 1,3-propanediol synthesis.
- This work paves the way for industrial applications of L. diolivorans in biorefining.
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