GABA-producing Lactic Acid Bacteria Research Articles

Lactococcus lactis CNCM I-5388 versus NCDO2118 by its GABA hyperproduction ability, counteracts faster stress-induced intestinal hypersensitivity in rats.

Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder characterized by its main symptom, visceral hypersensitivity (VH), which is aggravated by stress. Gut-brain interactions and gut bacteria may alleviate IBS symptoms, including VH. γ-amino butyric acid (GABA), produced notably by lactic acid bacteria (LAB), shows promising result in IBS symptoms treatment. In bacteria, GABA is generated through glutamate decarboxylase (GAD) metabolism of L-glutamic acid, maintaining intracellular pH. In mammals, GABA acts as an inhibitory neurotransmitter, modulating pain, stress, and anxiety. Therefore, utilizing GABA-producing LAB as a therapeutic approach might be beneficial. Our previous work showed that a GABA-producing Lactococcus lactis strain, NCDO2118, reduced VH induced by acute stress in rats after a 10-day oral treatment. Here, we identified the strain CNCM I-5388, with a four-fold higher GABA production rate under the same conditions as NCDO2118. Both strains shared 99.1% identical GAD amino acid sequences and invitro analyses revealed the same optimal pH for GAD activity; however, CNCM I-5388 exhibited 17 times higher intracellular GAD activity and increased resistance to acidic pH. Additionally, invivo experiments have demonstrated that CNCM I-5388 has faster anti-VH properties in rats compared with NCDO2118, starting from the fifth day of treatment. Finally, CNCM I-5388 anti-VH effects partially persisted after 5-day treatment interruption and after a single oral treatment. These findings highlight CNCM I-5388 as a potential therapeutic agent for managing VH in IBS patients.

Gamma-aminobutyric acid (GABA) promotes characteristics of Levilactobacillus sp. LB-2

γ-Aminobutyric acid (GABA) is a pharmaceutical, bioactive amino acid that can be produced by lactic acid bacteria (LAB), but the yield of GABA is still limited by several reasons. In this study, a GABA-producing LAB strain Levilactobacillus sp. LB-2 (OP890629) was isolated from a Chinese traditional dough fermentation starter of Jiaozi. When subjected to GABA production optimization applying one-factor-at-a-time (OFAT) and response surface method (RSM), the temperature, pH, and L-monosodium glutamate (L-MSG) concentration for GABA production were set as 37 °C, pH 4.5, and 2.5 g/L, respectively. The maximum amount of GABA produced by strain Levilactobacillus sp. LB-2 was 49.42 g/L, which was 3.5-fold higher than that before optimization of 14.19 g/L. The GABA content of steamed bread made by Levilactobacillus sp. LB-2 significantly increased. Thus, strain Levilactobacillus sp. LB-2 could be used for the production of GABA or applied for the development and improvement of GABA-enriched foods.

Selection and identification of antibacterial and GABA-producing lactic acid bacteria from Vietnamese fermented meat

This study aimed to screen the lactic acid bacteria (LAB) strain that has high potential in gamma-aminobutyric acid (GABA) production and antibacterial properties, which could be applied in Vietnamese fermented meat (Nem chua) fermentation. These bacterial strains were isolated from commercial Nem chua products and stored in the laboratory. GABA synthesized from 18 strains of LAB in the MRS media adding 3% MSG was qualitative and quantitative analyzed through thin layer chromatography (TLC) and its calibration curve equation, then antibacterial activity and lactic acid production were also tested by acidic titration method, and well diffusion method, respectively. The results showed that all LAB strains synthesized GABA with a concentration from 1.323 - 1.582 mg/mL; however, eight strains were recorded with higher potential, including VL1, XP1, TP1, HC2, CP2, LV2, TX7, and HK 162. These eight strains showed the inhibition zone to Escherichia coli indicator from 4.67 - 11.67 mm. Besides, they are capable of yielding lactic acid from 18.00 - 19.35 g/L after seven days of fermentation in MRS media. The strain VL1 with the highest potential was identified as Lactobacillus plantarum. This strain was suggested as a promising starter to be applied in the Nem chua fermentation process.

Probiotic Properties and Optimization of Gamma-Aminobutyric Acid Production by Lactiplantibacillus plantarum FBT215.

Gamma-aminobutyric acid (GABA) improves various physiological illnesses, including diabetes, hypertension, depression, memory lapse, and insomnia in humans. Therefore, interest in the commercial production of GABA is steadily increasing. Lactic acid bacteria (LAB) have widely been reported as a GABA producer and are safe for human consumption. In this study, GABA-producing LAB were preliminarily identified and quantified via GABase assay. The acid and bile tolerance of the L. plantarum FBT215 strain were evaluated. The one-factor-at-a-time (OFAT) strategy was applied to determine the optimal conditions for GABA production using HPLC. Response surface methodology (RSM) with Box-Behnken design was used to predict the optimum GABA production. The strain FBT215 was shown to be acid and bile tolerant. The optimization of GABA production via the OFAT strategy resulted in an average GABA concentration of 1688.65 ± 14.29 μg/ml, while it was 1812.16 ± 23.16 μg/ml when RSM was applied. In conclusion, this study provides the optimum culture conditions for GABA production by the strain FBT215 and indicates that L. plantarum FBT215 is potentially promising for commercial functional probiotics with health claims.

Gamma-aminobutyric acid fermentation in MRS-based medium by the fructophilic Lactiplantibacillus plantarum Y7.

Among the key metabolites produced by probiotic lactic acid bacteria (LAB), the use of gamma-aminobutyric acid (GABA), which alleviates hypertension, depression, and sleepiness in humans, is gaining popularity. Thus, GABA-producing LAB are sought after. GABA-producing LAB were preliminarily screened in acidified-MRS broth and quantified via GABase assays. The one-factor-at-a-time strategy was applied to determine the optimal conditions for GABA production. GABA production in reconstituted skim milk medium (RSM) and antibiotic susceptibility testing were performed to evaluate the potential of the strain as a yogurt starter. L. plantarum Y7 produced 4,856.86 ± 82.47μg/mL of GABA at optimal culture conditions. Co-cultivation of Y7 and commercial Lactobacillus bulgaricus affected the amount of GABA production (6.85 ± 0.20μg/mL) in RSM. Y7 was susceptible to ampicillin, erythromycin, and tetracycline. Therefore, L. plantarum Y7 represents a promising strain for GABA production in the food industry.

Screening of gamma-aminobutyric acid-producing lactic acid bacteria and its application in Monascus-fermented rice production.

Gamma-aminobutyric acid (GABA), with an antidepressant effect, and Monacolin K, with a cholesterol-lowering effect, are the main bioactive ingredients in Monascus-fermented rice (MFR). The simultaneous enrichment of both ingredients can effectively enhance the health benefits of MFR. However, the capacity of Monascus spp. to produce GABA is limited. Seventeen lactic acid bacteria (LAB) strains were preliminarily screened for GABA-producing by whole-cells bioconversion of L-glutamate, followed by rescreening through fermentation with the addition of the precursor L-glutamic acid. Subsequently, the bioconversion conditions (temperature, metal ions, and pH) for the conversion of L-monosodium glutamate (MSG) were investigated. Additionally, the GABA-producing LAB was co-inoculated with a monacolin K producing strain Monascus anka 20-2, and the ratio of M. anka 20-2 to LAB in microbial consortia was optimized for MFR production. The strain Lactobacillus plantarum 8014 was screened out for its ability to produce GABA. At an optimal temperature of 33°C and pH 7.5, with the addition of 0.05 g/L ZnSO4, the strain showed an L-glutamate conversion rate of 100%. The ratio optimization of M. anka 20-2 to L. plantarum 8014 in microbial consortia showed that when the dry cell ratio was 2:1, the content of monacolin K and GABA in the MFR simultaneously reached 2.22 mg/g and 29.9 mg/g, respectively. A two-stage fermentation using microbial consortia containing M. anka 20-2 and L. plantarum 8014 was developed for the production of bioactive MFR, in which the active ingredients monacolin K and GABA were simultaneously enriched, with good consumer acceptability due to the aromatic scent produced by lactic acid bacteria.

Screening of GABA-Producing Lactic Acid Bacteria from Thai Fermented Foods and Probiotic Potential of Levilactobacillus brevis F064A for GABA-Fermented Mulberry Juice Production

Gamma-aminobutyric acid (GABA), the inhibitory neurotransmitter, can be naturally synthesized by a group of lactic acid bacteria (LAB) which is commonly found in rich carbohydrate materials such as fruits and fermented foods. Thirty-six isolates of GABA-producing LAB were obtained from Thai fermented foods. Among these, Levilactobacillus brevis F064A isolated from Thai fermented sausage displayed high GABA content, 2.85 ± 0.10 mg/mL and could tolerate acidic pH and bile salts indicating a promising probiotic. Mulberry (Morus sp.) is widely grown in Thailand. Many mulberry fruits are left to deteriorate during the high season. To increase its value, mulberry juice was prepared and added to monosodium glutamate (MSG), 2% (w/v) prior to inoculation with 5% (v/v) of L. brevis F064A and incubated at 37 °C for 48 h to obtain the GABA-fermented mulberry juice (GABA-FMJ). The GABA-FMJ obtained had 3.31 ± 0.06 mg/mL of GABA content, 5.58 ± 0.52 mg gallic acid equivalent/mL of antioxidant activity, 234.68 ± 15.53 mg cyanidin-3-glucoside/mL of anthocyanin, an ability to inhibit growth of Bacillus cereus TISTR 687, Salmonella Typhi DMST 22842 and Shigella dysenteriae DMST 1511, and 10.54 ± 0.5 log10 colony-forming units (CFU)/mL of viable L. brevis F064A cell count. This GABA-FMJ was considered as a potential naturally functional food for human of all ages.

Glutamate Decarboxylase from Lactic Acid Bacteria-A Key Enzyme in GABA Synthesis.

Glutamate decarboxylase (l-glutamate-1-carboxylase, GAD; EC 4.1.1.15) is a pyridoxal-5’-phosphate-dependent enzyme that catalyzes the irreversible α-decarboxylation of l-glutamic acid to γ-aminobutyric acid (GABA) and CO2. The enzyme is widely distributed in eukaryotes as well as prokaryotes, where it—together with its reaction product GABA—fulfils very different physiological functions. The occurrence of gad genes encoding GAD has been shown for many microorganisms, and GABA-producing lactic acid bacteria (LAB) have been a focus of research during recent years. A wide range of traditional foods produced by fermentation based on LAB offer the potential of providing new functional food products enriched with GABA that may offer certain health-benefits. Different GAD enzymes and genes from several strains of LAB have been isolated and characterized recently. GABA-producing LAB, the biochemical properties of their GAD enzymes, and possible applications are reviewed here.

김치로부터 Gamma-Aminobutyric Acid(GABA)를 생산하는 유산균의 분리 및 특성

To develop Gamma-aminobutyric acid(GABA)-producing probiotics we isolated GABA-producing lactic acid bacteria from kimchi. 17 strains of GABA-producing lactic acid bacteria were isolated among 703 lactic acid bacteria originated from 21 kind of kimchi. Two strains, K991 and K998, were selected with highest GABA production ability. The results of 16S rRNA gene sequence analysis, K991 and K998 were identified as Lactococcus lactis, which showed 99.93% and 99.87% similarity, respectively. When the two strains were cultivated in MRS containing 5% MSG at 30°C, GABA began to be produced after 12 hours, reaching maximum production at 40 hours(13.20 mM and 13.35 mM, respectively). In contrast, GABA production was about three times lower when MSG was not added.

Gamma-aminobutyric acid (GABA) production in milk fermented by specific wild lactic acid bacteria strains isolated from artisanal Mexican cheeses

PurposeThe purpose of this study was to screen wild GABA-producing lactic acid bacteria (LAB) isolated from artisanal Mexican cheeses and to evaluate the fermentation conditions for the enhancement of the GABA yield in fermented milk.MethodsA qualitative test was carried out to select the GABA-producing LAB and the GABA was quantified by reversed-phase high-performance liquid chromatography in fermented milk (FM). Two inoculum concentrations (107 and 109 CFU/mL), two incubation temperatures (30 and 37 °C), three glutamate concentrations (1, 3, and 5 g/L), and three pyridoxal 5′-phosphate (PLP) concentrations (0, 100, and 200 μM) were assessed to establish suitable conditions to enhance the GABA yield in FM.ResultsResults showed that, from a total of 94 LAB strains, fermented milk with two Lactococcus lactis strains (L-571 or L-572) presented the highest GABA production. However, 37 °C of incubation and 109 CFU/mL and 3 g/L of glutamate significantly led the highest GABA yield in FM with L-571. Further studies are needed to establish the optimum conditions for producing GABA by this strain, and in vivo studies may reveal its potential use as GABA-producing culture.ConclusionThese results highlight the importance of wild LAB strains in order to generate new alternatives and opportunities in the development of functional foods containing GABA.

Production of Gamma-Aminobutyric Acid from Lactic Acid Bacteria: A Systematic Review.

Gamma-aminobutyric acid (GABA) is widely distributed in nature and considered a potent bioactive compound with numerous and important physiological functions, such as anti-hypertensive and antidepressant activities. There is an ever-growing demand for GABA production in recent years. Lactic acid bacteria (LAB) are one of the most important GABA producers because of their food-grade nature and potential of producing GABA-rich functional foods directly. In this paper, the GABA-producing LAB species, the biosynthesis pathway of GABA by LAB, and the research progress of glutamate decarboxylase (GAD), the key enzyme of GABA biosynthesis, were reviewed. Furthermore, GABA production enhancement strategies are reviewed, from optimization of culture conditions and genetic engineering to physiology-oriented engineering approaches and co-culture methods. The advances in both the molecular mechanisms of GABA biosynthesis and the technologies of synthetic biology and genetic engineering will promote GABA production of LAB to meet people’s demand for GABA. The aim of the review is to provide an insight of microbial engineering for improved production of GABA by LAB in the future.

Identification, Classification and Screening for γ-Amino-butyric Acid Production in Lactic Acid Bacteria from Cambodian Fermented Foods.

Screening for various types of lactic acid bacteria (LAB) that form the biological agent γ-amino-butyric acid (GABA) is important to produce different kinds of GABA-containing fermented foods. So far, no GABA-producing LAB have been reported from Cambodian fermented foods. Most small-scale fermentations and even some industrial processes in this country still rely on indigenous LAB. The application of GABA-producing autochthonous starters would allow the production of Cambodian fermented foods with an additional nutritional value that meet the population’s dietary habits and that are also more attractive for the international food market. Matrix-assisted laser desorption/ionizing time-of-flight mass spectrometry (MALDI-TOF MS) and partial 16S rDNA sequencing were used to identify 68 LAB isolates from Cambodian fermented foods. These isolates were classified and grouped with (GTG)5 rep-PCR, resulting in 50 strains. Subsequently, all strains were investigated for their ability to produce GABA by thin layer chromatography. GABA-positive strains were further analyzed by the GABase assay. Of the six GABA-positive LAB strains—one Lactobacillus futsaii, two Lactobacillus namurensis, and three Lactobacillus plantarum strains—two Lactobacillus plantarum strains produced high amounts of GABA (20.34 mM, 16.47 mM). These strains should be further investigated for their potential application as GABA-producing starter cultures in the food applications.

Molecular identification and phylogenetic analysis of GABA-producing lactic acid bacteria isolated from indigenous dadih of West Sumatera, Indonesia.

Background: Dadih (fermented buffalo milk) is a traditional Indonesian food originating from West Sumatra province. The fermentation process is carried out by lactic acid bacteria (LAB), which are naturally present in buffalo milk. Lactic acid bacteria have been reported as one of potential producers of γ-aminobutyric acid (GABA). GABA acts as a neurotransmitter inhibitor of the central nervous system. Methods: In this study, molecular identification and phylogenetic analysis of GABA producing LAB isolated from indigenous dadih of West Sumatera were determined. Identification of the GABA-producing LAB DS15 was based on conventional polymerase chain reaction. 16S rRNA gene sequence analysis was used to identify LAB DS15. Results: PCR of the 16S rRNA gene sequence of LAB DS15 gave an approximately 1400 bp amplicon. Phylogenetic analysis showed that LAB DS15 was Pediococcus acidilactici, with high similarity of 99% at 100% query coverage to Pediococcus acidilactici strain DSM 20284. Conclusions: It can be concluded that GABA producing LAB isolated from indigenous dadih was Pediococcus acidilactici.

Laktik Asit Bakterileri Tarafından Üretilen Gamma Aminobütirik Asitin Önemi

Gamma aminobütirik asit (GABA), insan beynindeki önemli bir inhibitör olan nörotransmitter olarak bilinmektedir. Son yıllarda yapılmış çalışmalar GABA‘nın sağlık üzerine birçok yararlı etkisi olduğunu göstermektedir. GABA takviyesi kullanan insanlarda, psikolojik sorunlar, uykusuzluk ve bağışıklık problemleri gibi hayatı olumsuz yönde etkileyen faktörlerin azaldığı bildirilmiştir. Bu alanda yapılan çalışmalar laktik asit bakterileri (LAB)‘nin GABA üretebildiğini göstermiştir. Bu sebeple GABA üreten LAB‘a olan ilgi giderek artmaktadır. LAB tarafından doğal yolla üretilen GABA'nın sentetik olarak üretilenlere kıyasla yan etkilerinin neredeyse hiç olmadığı belirtilmektedir. Özellikle Uzak Doğu ülkelerindeki gıdalardan izole edilmiş GABA üreten LAB‘ın fonksiyonel gıda geliştirmede kullanılması da önerilmektedir. Yeni gelişmekte olan bu alanda ilerleyebilmek için daha çok araştırma yapılması ve GABA‘nın keşfedilmemiş özelliklerinin belirlenmesi gerekmektedir. Bu derlemede GABA‘nın öneminden bahsedilmiş ve GABA ile ilgili yapılmış çalışmalar üzerinde durulmuştur.

Isolation and Characterization of Lactic Acid Bacteria Producing GABA from Indigenous West Sumatera Fermented Food

The purpose of this study was to isolate and characterize lactic acid bacteria (LAB) from indigenous West Sumatera fermented foods. The aim was to obtain LAB isolates that have the ability to produce γ-aminobutyric acid (GABA) and to characterize the highest GABA producing LAB. The indigenous West Sumatera fermented foods sampled were dadih (fermented buffalo milk), asam durian (fermented durian), tape singkong (fermented cassava) and ikan budu (fermented fish). The conventional method was used to isolate LAB, after which LAB were screened for γ-amino butyric acid (GABA) and GABA producing LAB characterized by biochemical tests. A total of 704 isolates were successfully isolated. Five hundred fifty two (552) isolates were identified as LAB and 103 isolates were confirmed as GABA producing LAB. The highest GABA producing LAB was DS15 and produced 49.365 mg/ml of GABA. DS18 produced the least GABA of 3.415 mg/ml. Biochemical assay of DS15 showed that the isolate could grow on the MRSA, undergoes aerobic respiration, do not use lactose, glucose, mannitol, arginine, aesculin, arabinose, raffinose, sorbitol, trehalase and xylose/melezitose. The isolate DS15 was also negative for arginine, nitrate reduction, catalase and oxidase tests. Based on the gram stain reaction, the isolate DS15 was a gram-positive bacterium and could be classified as Lactobacillus sp.

Molecular identification and phylogenetic analysis of GABA-producing lactic acid bacteria isolated from indigenous dadih of West Sumatera, Indonesia

Background: Dadih (fermented buffalo milk) is a traditional Indonesian food originating from West Sumatra province. The fermentation process is carried out by lactic acid bacteria (LAB), which are naturally present in buffalo milk. Lactic acid bacteria have been reported as one of potential producers of γ-aminobutyric acid (GABA). GABA acts as a neurotransmitter inhibitor of the central nervous system. Methods: In this study, molecular identification and phylogenetic analysis of GABA producing LAB isolated from indigenous dadih of West Sumatera were determined. Identification of the GABA-producing LAB DS15 was based on conventional polymerase chain reaction. 16S rRNA gene sequence analysis was used to identify LAB DS15. Results: PCR of the 16S rRNA gene sequence of LAB DS15 gave an approximately 1400 bp amplicon. Phylogenetic analysis showed that LAB DS15 was Pediococcus acidilactici, with high similarity of 99% at 100% query coverage to Pediococcus acidilactici strain DSM 20284. Conclusions: It can be concluded that GABA producing LAB isolated from indigenous dadih was Pediococcus acidilactici.

GABA-Producing Natural Dairy Isolate From Artisanal Zlatar Cheese Attenuates Gut Inflammation and Strengthens Gut Epithelial Barrier in vitro.

Probiotic bacteria are recognized for their health-promoting properties, including maintenance of gut epithelial integrity and host immune system homeostasis. Taking into account the beneficial health-promoting effects of GABA, the presence of the gadB gene, encoding glutamate decarboxylase that converts L-glutamate to GABA, was analyzed in Lactic Acid Bacteria (LAB) natural isolates from Zlatar cheese. The results revealed that 52% of tested Lactobacillus spp. and 8% of Lactococcus spp. isolates harbor the gadB gene. Qualitative and quantitative analysis of GABA production performed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) revealed the highest GABA production by Lactobacillus brevis BGZLS10-17. Since high GABA-producing LAB natural isolates are the most valuable source of naturally produced GABA, the probiotic properties of BGZLS10-17 were characterized. This study demonstrated high adhesion of BGZLS10-17 strain to Caco-2 cells and the ability to decrease the adhesion of Escherichia coli ATCC25922 and Salmonella enterica C29039. Treatment of differentiated Caco-2 cells monolayer with BGZLS10-17 supernatant containing GABA alleviated inflammation (production of IL-8) caused by IL-1β and significantly stimulated the expression of tight junction proteins (zonulin, occludin, and claudin 4), as well as the expression of TGF-β cytokine leading to the conclusion that immunosuppression and strengthening the tight junctions can have significant role in the maintenance of intestinal epithelial barrier integrity. Taken together the results obtained in this study support the idea that using of GABA producing BGZLS10-17 probiotic strain could be a good strategy to modulate immunological response in various inflammatory diseases, and at the same time, it could be a good candidate for adjunct starter culture for production of GABA-enriched dairy foods and beverages offering new perspectives in designing the novel functional foods.

Molecular identification and phylogenetic analysis of GABA-producing lactic acid bacteria isolated from indigenous dadih of West Sumatera, Indonesia

Background: Dadih (fermented buffalo milk) is a traditional Indonesian food originating from West Sumatra province. The fermentation process is carried out by lactic acid bacteria (LAB), which are naturally present in buffalo milk. Lactic acid bacteria have been reported as one of potential producers of γ-aminobutyric acid (GABA). GABA acts as a neurotransmitter inhibitor of the central nervous system. Methods: In this study, molecular identification and phylogenetic analysis of GABA producing LAB isolated from indigenous dadih of West Sumatera were determined. Identification of the GABA-producing LAB DS15 was based on conventional polymerase chain reaction. 16S rRNA gene sequence analysis was used to identify LAB DS15. Results: PCR of the 16S rRNA gene sequence of LAB DS15 gave an approximately 1400 bp amplicon. Phylogenetic analysis showed that LAB DS15 was Pediococcus acidilactici, with high similarity of 99% at 100% query coverage to Pediococcus acidilactici strain DSM 20284. Conclusions: It can be concluded that GABA producing LAB isolated from indigenous dadih was Pediococcus acidilactici.

Molecular identification and phylogenetic analysis of GABA-producing lactic acid bacteria isolated from indigenous dadih of West Sumatera, Indonesia

Background: Dadih (fermented buffalo milk) is a traditional Indonesian food originating from West Sumatra province. The fermentation process is carried out by lactic acid bacteria (LAB), which are naturally present in buffalo milk. Lactic acid bacteria have been reported as one of potential producers of γ-aminobutyric acid (GABA). GABA acts as a neurotransmitter inhibitor of the central nervous system. Methods: In this study, molecular identification and phylogenetic analysis of GABA producing LAB isolated from indigenous dadih of West Sumatera were determined. Identification of the GABA-producing LAB DS15 was based on conventional polymerase chain reaction. 16S rRNA gene sequence analysis was used to identify LAB DS15. Results: PCR of the 16S rRNA gene sequence of LAB DS15 gave an approximately 1400 bp amplicon. Phylogenetic analysis showed that LAB DS15 was Pediococcus acidilactici, with high similarity of 99% at 100% query coverage to Pediococcus acidilactici strain DSM 20284. Conclusions: It can be concluded that GABA producing LAB isolated from indigenous dadih was Pediococcus acidilactici.

Properties of Kimchi Fermented with GABA-Producing Lactic Acid Bacteria as a Starter.

Kimchi (a traditional Korean fermented vegetable) was prepared with a starter, Lactobacillus zymae GU240 producing γ-aminobutyric acid (GABA), and one precursor of GABA (glutamic acid, glutamic acid monosodium salt (MSG), or kelp extract). L. zymae GU240, an isolate from kimchi, can grow at 7% NaCl and low temperature. Five different kimchi samples were fermented for 20 weeks at -1°C. Kimchi with starter alone could not produce GABA. The GABA content was highest in kimchi with co-inoculation of the starter and MSG (1% (w/w)). Kimchi co-inoculated with the starter and kelp extract powder (3% (w/w)) had the second highest GABA content. Addition of glutamic acid powder (1% (w/w)) caused a reduction in the pH level of kimchi and growth inhibition of lactic acid bacteria and yeasts. Kimchi samples with MSG or kelp extract showed improvement of sensory evaluation scores. The results demonstrate the possibility to produce kimchi with improved functionality and taste by using L. zymae GU240 as a starter along with a suitable precursor such as MSG or kelp extract.