Abstract
Objectives
This study aimed to screen, characterize, and annotate the genome along with the comparison of GABA synthesis genes presented in lactic acid bacteria (LAB).
Results
Thirty-five LAB isolates from fermented foods were screened for GABA production using thin-layer chromatography (TLC). Fifteen isolates produced GABA ranging from 0.07 to 22.94 g/L. Based on their GTG5 profiles, phenotypic, and genotypic characteristics, isolates LSI1-1, LSI1-5, LSI2-1, LSI2-2, LSI2-3, LSI2-5, and LSM3-1-4 were identified as Lactobacillus plantarum subsp. plantarum; isolate LSM1-4 was Lactobacillus argentoratensis; isolates CAB1-2, CAB1-5, CAB1-7, and LSI1-4 were Lactobacillus pentosus; and CAB1-1, LSM3-1-1 and LSM3-2-3 were Lactobacillus fermentum. Strains LSI2-1 and CAB1-7 from pickled vegetables were selected for genome analysis. The gadA gene (1410 bp, 470aa) was encountered in GABA production of both strains and no other glutamate decarboxylase (GAD) genes were found in the genomes when compared with other LAB strains. The presence of gadA is evidence for GABA production. Strains LSI2-1 and CAB1-7 produced 22.94 g/L and 11.59 g/L of GABA in GYP broth supplemented with 3% (w/v) MSG at 30 °C for 72 h, respectively.
Conclusions
Our report highlights the characterization of LAB and GABA production of L. plantarum LSI2-1 strain with its GABA synthesis gene.
Graphic abstract
GABA production of strains LSI2-1 and CAB1-7 in GYP broth with 3% (w/v) MSG and comparative GAD genes
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Acknowledgements
We thank Dr. Nattakorn Kuncharoen for genome sequence analysis, and Mr. Engkarat Kingkaew and Ms. Saranporn Poothong for 16S rRNA gene sequence analysis.
Supplementary Information
Supplementary Fig. S1—Calibration curve of GABA concentration in the range of 0–16 μg/mL.
Supplementary Fig. S2—Chromatograms of MSG and GABA analysis using HPLC.
Supplementary Fig. S3—A genome circular maps of strain LSI2-1 (a) and CAB1-7 (b) created by the ‘circular viewer’ functionality implemented in the PATRIC web server.
Supplementary Table S1—ANOVA table showed the efficiency on GABA production of isolates.
Supplementary Table S2—The multiple comparisons of GABA production of isolates using Tukey’s multiple range tests.
Funding
This research was supported by a Grant for International Research Integration: Research Pyramid, Ratchadaphiseksomphot Endowment Fund (GCURP_58_01_33_01), and the Ratchadapiseksomphot Endowment Fund, Chulalongkorn University for a Postdoctoral Fellowship to Dr. Sukanya Phuengjayaem.
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SP performed the isolation, screening, identification, genome sequence analysis and writing the manuscript, AB contributed to the GTG5 profiles, and ST performed the identification and writing the manuscript. All authors read and approved the final manuscript.
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Phuengjayaem, S., Booncharoen, A. & Tanasupawat, S. Characterization and comparative genomic analysis of gamma-aminobutyric acid (GABA)-producing lactic acid bacteria from Thai fermented foods. Biotechnol Lett 43, 1637–1648 (2021). https://doi.org/10.1007/s10529-021-03140-y
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DOI: https://doi.org/10.1007/s10529-021-03140-y