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Production of Organic Acids

Production of Organic Acids

Lactic acid, an important organic acid extensively applied in food, pharmaceutical, and biodegradable polymer industries, is primarily produced via microbial fermentation. This method is favored over chemical synthesis due to its environmental sustainability and capacity for enantiomerically pure product formation. Among various microbial processes, the fermentation of starch-based substrates stands out due to the abundance and renewability of raw materials like corn and potatoes.Hydrolysis of...

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Related Experiment Video

Updated: Apr 17, 2026

Author Spotlight: Methods for Electroporation and Transformation Confirmation in Limosilactobacillus reuteri DSM20016

Author Spotlight: Methods for Electroporation and Transformation Confirmation in Limosilactobacillus reuteri DSM20016

Published on: June 23, 2023

High efficiency electrotransformation of Lactobacillus casei.

Dennis L Welker¹, Joanne E Hughes², James L Steele³

¹Department of Biology, Utah State University, Logan, UT 84322-5305, USA dennis.welker@usu.edu.

FEMS Microbiology Letters

|February 12, 2015

Summary

Optimized electrotransformation protocols enhance gene transfer in Lactobacillus casei strains. These improved methods achieve high transformation efficiencies, enabling genetic manipulation for research and development.

Keywords:

electroporation genetic manipulation lactic acid bacteria transformation

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

High-efficiency electrotransformation is crucial for genetic manipulation of lactic acid bacteria.
Existing protocols for other species may not directly apply to Lactobacillus casei strains.

Purpose of the Study:

To evaluate and optimize electrotransformation protocols for five distinct Lactobacillus casei strains.
To determine the applicability of established high-efficiency electrotransformation methods to Lactobacillus casei.

Main Methods:

Investigated the impact of cell growth conditions (glycine, NaCl), culture density (OD600), and pre-electroporation treatments (water, LiAc/DTT) on transformation efficiency.
Optimized electroporation parameters for Lactobacillus casei.
Assessed transformation efficiencies using pTRKH2 DNA.

Main Results:

Specific treatments significantly improved electrotransformation efficiencies in Lactobacillus casei.
Strain-specific variations in response to optimization protocols were observed.
Achieved transformation efficiencies of 10^6 colony-forming units per μg of pTRKH2 DNA in three strains.

Conclusions:

Established electrotransformation protocols can be adapted for efficient genetic manipulation of Lactobacillus casei.
The achieved efficiencies are sufficient for constructing chromosomal gene knock-outs and gene replacements.
Further optimization may be required due to strain-dependent variability.