Table S1. Measurement of glutamate and GABA in Levilactobacillus brevis Lbr-6108, Lbr-35, and ATCC 14869. The p-value is the result of an ANOVA with Tukey's multiple comparison test comparing the ODs of each strain to at T48 to Lbr-6108 in the respective media.
StrainTimepoint (hr)rep 1rep 2rep 1rep 2rep 1rep 2p-valuerep 1rep 2rep 1rep 2rep 1rep 2p-valueLbr-6108066067050500.1390.1418900880050500.1370.133Lbr-6108365064050500.4340.4329000860060580.4120.397
Lbr-6108662059066700.6670.671860086001001000.6150.602Lbr-610895705102101901.0561.069870085003103200.9590.968Lbr-6108123603503403501.2911.320810081006206301.2171.226Lbr-6108151301205005201.4781.44872007600120012001.4061.386Lbr-6108181001005005001.6381.61163006300190020001.5421.540Lbr-6108211001005005001.7671.76350005100280028001.6891.722Lbr-6108241001005005001.8161.83337003700400038001.7891.774Lbr-6108481001005005101.7331.753250300600060001.7241.733Lbr-35069067050500.1420.1368900880053500.1450.139Lbr-35368066050500.5190.5008900900050500.4760.491
Lbr-35667064050500.8550.8498800860050500.7800.753Lbr-35967070054591.0861.1108600890076701.0291.021Lbr-351271068058501.2781.2718600900061561.1591.144Lbr-351569068050511.4461.3978800890075601.2661.278Lbr-351868069051521.5561.5448700890063631.3971.401Lbr-352167068054581.6501.6878900950071721.5021.564Lbr-352463060082801.7411.738870088001201101.5661.579Lbr-35485605401701601.6901.690>0.9999900089002702301.6351.621>0.9999 glutamate, MRS (μg/mL)Glutamate, MRS+MSG (μg/mL)GABA, MRS (μg/mL)OD, MRS (μg/mL)GABA, MRS+MSG (μg/mL)OD, MRS+MSG (600nm)
Table S2. Glutamate utilization and GABA production by Levilactobacillus brevis Lbr-6108 in MRS with different concentrations of MSG.
Timepoint (hr)rep 1rep 2rep 1rep 2rep 1rep 2rep 1rep 2rep 1rep 2rep 1rep 2rep 1rep 2rep 1rep 2rep 1rep 25500.0450.050.050.010.0%11.1%1000.0340.050.050.05.0%14.7%5000.04740.050.050.01.0%1.1%244200.04290.02600.02620.061.9%61.1%6000.05920.02900.02930.048.3%49.5%4000.03790.01800.01830.045.0%48.3%488200.08160.05300.05310.064.6%65.1%16300.016270.010000.09970.061.3%61.3%24000.023530.013000.012680.054.2%53.9%728000.07980.06100.06075.076.3%76.1%18200.018405.012000.012140.065.9%66.0%28000.023530.016000.016450.057.1%69.9% MRS with 10,000 μg/mL MSGMRS with 30,000 μg/mL MSGMRS with 90,000 μg/mL MSGGlu Uptake (μg/mL)GABA (μg/mL)% YieldGlu Uptake (μg/mL)GABA (μg/mL)% YieldGlu Uptake (μg/mL)GABA (μg/mL)% Yield
Table S3. Gene List from Figure 4. Locus IDs are for Levilactobacillus brevis Lbr-6108 genome.
Encoded protein functions are given for genes shown in Figure 4.
Locus ID Gene
name Protein function Operon / pathway
Panel in Figure
4
BGC39_04180 gadA Glutamate decarboxylase (EC 4.1.1.15) GadA
L-Glutamate decarboxylase
operon A
BGC39_07245 gadR Transcriptional regulator GadR
L-Glutamate decarboxylase
operon A
BGC39_07250 gadC Glutamate/gamma- aminobutyrate antiporter
L-Glutamate decarboxylase
operon A
BGC39_07255 gadB Glutamate decarboxylase (EC 4.1.1.15) GadB
L-Glutamate decarboxylase
operon A
BGC39_07260 gltX Glutamyl-tRNA synthetase (EC 6.1.1.17; EC 6.1.1.24)
L-Glutamate decarboxylase
operon A
BGC39_05810 hmpT
Substrate-specific component HmpT of predicted
hydroxymethylpyrimidine ECF transporter
Pyridoxal 5’-phosphate (PLP) B
BGC39_05815 thiD2 Novel pyridoxal kinase, thiD
family (EC 2.7.1.35) Pyridoxal 5’-phosphate (PLP) B
BGC39_05820 norD
Transcriptional regulator of pyridoxine metabolism / Pyridoxamine phosphate aminotransferase (EC 2.6.1.54)
Pyridoxal 5’-phosphate (PLP) B
BGC39_05050 argF
Ornithine
carbamoyltransferase (EC 2.1.3.3)
Arginine deiminase operon C BGC39_05055 arcA Arginine deiminase (EC
3.5.3.6) Arginine deiminase operon C
BGC39_05060 arcD Arginine/ornithine antiporter
ArcD Arginine deiminase operon C
BGC39_06170 aguA Agmatine deiminase (EC
3.5.3.12) Agmatine deiminase operon D
BGC39_06175 aguD
Agmatine/putrescine antiporter, associated with agmatine catabolism
Agmatine deiminase operon D
BGC39_06180 aguB
Putrescine
carbamoyltransferase (EC 2.1.3.6)
Agmatine deiminase operon D
BGC39_06135 mleS Malolactic enzyme (EC
4.1.1.101) Malolactic fermentation operon E BGC39_06140 mleR Malolactic regulator MleR Malolactic fermentation operon E BGC39_06185 nhaC2 Predicted tyrosine transporter,
NhaC family Tyrosine decarboxylase operon F BGC39_06190 tdcP Predicted tyrosine transporter,
GadC family Tyrosine decarboxylase operon F BGC39_06195 tdcA L-tyrosine decarboxylase (EC
4.1.1.25) TDC Tyrosine decarboxylase operon F BGC39_06200 tyrS Tyrosyl-tRNA synthetase (EC
6.1.1.1) Tyrosine decarboxylase operon F
Figure S1. Culture purity testing by species- and strain-specific PCR for Levilactobacillus brevis Lbr-6108 and Lbr-35.
Figure S2. PCA plot of RNA-sequencing for Levilactobacillus brevis Lbr-6108 and Lbr-35.
