Abstract
This study was conducted to investigate the γ-aminobutyric acid (GABA) production ability of 20 Lactobacillus and 25 Bifidobacterium strains which were previously isolated in our laboratory. Effect of initial pH, incubation time, monosodium glutamate (MSG), and pyridoxal-5′-phosphate (PLP) concentration for highest GABA production by two potent bacterial strains, Levilactobacillus brevis LAB6 and Limosilactobacillus fermentum LAB19 were optimized in the MRS media. A threefold increase in GABA production at an initial pH 4.0, incubation time of 120 h in medium supplemented with 3% MSG and 400 μM of PLP for LAB6 and 300 μM for LAB19 lead to the production of 19.67 ± 0.28 and 20.77 ± 0.14 g/L of GABA, respectively. Coculturing both strains under optimized conditions led to a GABA yield of 20.02 ± 0.17 g/L. Owing to potent anti-inflammatory activity in-vitro, as reported previously, and highest GABA production ability of LAB6 (MTCC 25662), its whole-genome sequencing and bioinformatics analysis was carried out for mining genes related to GABA metabolism. LAB6 harbored a complete glutamate decarboxylase (GAD) gene system comprising gadA, gadB, and gadC as well as genes responsible for the beneficial probiotic traits, such as for acid and bile tolerance and host adhesion. Comparative genomic analysis of LAB6 with 28 completely sequenced Levilactobacillus brevis strains revealed the presence of 95 strain-specific genes-families that was significantly higher than most other L. brevis strains.
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Data availability
The data sets generated during and/or analyzed during the current study are available from the corresponding author upon reasonable request. Whole genome sequencing data presented in this study are openly available in The National Centre for Biotechnology Information (NCBI).
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Acknowledgements
The authors would like to thank the Department of Biotechnology (DBT), Government of India and National Agri-Food Biotechnology Institute (NABI) for providing the research grant and the facilities. KK, KC, SRJ, and MB would like to acknowledge DBT project NERBPMC (BT/PR16088/NE/95/69/2015 NER-DBT) for providing the research grant. DeLCON (DBT e-library consortium) is highly acknowledged for providing access to the journals to NABI.
Funding
This research was funded by the Department of Biotechnology (DBT), Government of India, for the research grant given to the National Agri-Food Biotechnology Institute (NABI), Kanthi Kiran Kondepudi & Mahendra Bishnoi, Kanwaljit Chopra and Santa Ram Joshi.
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TM conducted the experiments and performed the whole-genome analysis and in-silico data analysis; RB isolation of Lactobacillus strains; KK, MB, KC, and SRJ provided the financial support for carrying out the research; KK, MB, and KC conceptualized, designed the experiments, edited the manuscript, and got funding to carry out the study. All authors have read and approved the final manuscript.
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Matta, T., Bhatia, R., Joshi, S.R. et al. GABA synthesizing lactic acid bacteria and genomic analysis of Levilactobacillus brevis LAB6. 3 Biotech 14, 62 (2024). https://doi.org/10.1007/s13205-024-03918-7
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DOI: https://doi.org/10.1007/s13205-024-03918-7