Co-Culture Optimization of Lactococcus lactis and Lactobacillus plantarum for Increasing GABA Production
Abstract
Gamma-aminobutyric acid (GABA) is a non-protein amino acid neurotransmitter that helps control nerve cell hyperactivity. Reduced GABA levels have been associated with symptoms of multiple sclerosis and treatment with GABAergic-associated agents has been shown in EAE mouse models to possibly alleviate symptoms and increase quality of life (1, 2). As with all medications, there is a risk of adverse side effects and low absorbance levels. Probiotics may be a better option and the creation of a custom probiotic requires experimentation and co-culturing. Two strains of GAD-Lactococcus lactis (GAD-L.lactis) designated P and P8, were each co-cultured with Lactobaccilus plantarum (L. plantarum), designated B and Corynebacterium glutamicum (C. glutamicum) designated G. These strains of GAD-L. lactis were created and provided through collaboration with Dr. Andrea Castillo from EWU and are part of a currently unpublished paper by Dr. Castillo (3). The growth curves for each bacterium were determined through OD readings taken at eight time points between 0 and 48 hrs. and compared to CFU counts at the same time points. The mid-log phase was found to determine the optimal growing time for adding the co-culture bacteria. Each bacterium was tested on its individual growing media and each other’s to determine the best media for the co-culture experiments. P+B and P8+B were able to grow in both M17 broth and TSB with results. P+G and P8+G only grew acceptably in TSB with results. All three combinations were successfully co-cultured, and supernatants were collected for future testing of glutamic acid and GABA levels.
Co-Culture Optimization of Lactococcus lactis and Lactobacillus plantarum for Increasing GABA Production
Gamma-aminobutyric acid (GABA) is a non-protein amino acid neurotransmitter that helps control nerve cell hyperactivity. Reduced GABA levels have been associated with symptoms of multiple sclerosis and treatment with GABAergic-associated agents has been shown in EAE mouse models to possibly alleviate symptoms and increase quality of life (1, 2). As with all medications, there is a risk of adverse side effects and low absorbance levels. Probiotics may be a better option and the creation of a custom probiotic requires experimentation and co-culturing. Two strains of GAD-Lactococcus lactis (GAD-L.lactis) designated P and P8, were each co-cultured with Lactobaccilus plantarum (L. plantarum), designated B and Corynebacterium glutamicum (C. glutamicum) designated G. These strains of GAD-L. lactis were created and provided through collaboration with Dr. Andrea Castillo from EWU and are part of a currently unpublished paper by Dr. Castillo (3). The growth curves for each bacterium were determined through OD readings taken at eight time points between 0 and 48 hrs. and compared to CFU counts at the same time points. The mid-log phase was found to determine the optimal growing time for adding the co-culture bacteria. Each bacterium was tested on its individual growing media and each other’s to determine the best media for the co-culture experiments. P+B and P8+B were able to grow in both M17 broth and TSB with results. P+G and P8+G only grew acceptably in TSB with results. All three combinations were successfully co-cultured, and supernatants were collected for future testing of glutamic acid and GABA levels.