Study on conditions for Gama buryric acid (GABA) production in Lactobacillus fermentum A01 isolated from human

Author(s):  
Cat Thien Phuc ◽  
Doan Thi Thanh Vinh ◽  
Truong Khoa My ◽  
Tran Thi Thu Hien ◽  
Nguyen Anh Dung ◽  
...  

Lactic acid bacteria play a vital role in biosynthesis of γ -aminobutyric acid (GABA) in the presence of glutamic acid - major substrate for the process. In recent study, Lactobacillus fermentum A01 (L. fermentum A01) isolated from various sources in Vietnam were screened for bacteria strains with high efficiency in GABA formation. L. fermentum was cultured in MRS broth containing 25 mg/mL monosodium glutamate (MSG), at pH of 6.5 and incubated at the optimal conditions (37ºC, for 24, 48, 72 h). After extraction and thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) analysis, L. fermentum A01 showed the GABA yield about 1.34 mg/g in dried supernatant, suggesting L. fermentum A01 to be a promising GABA producer for food and pharmaceutical applications.

Processes ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 22
Author(s):  
Ida Bagus Agung Yogeswara ◽  
Suwapat Kittibunchakul ◽  
Endang Sutriswati Rahayu ◽  
Konrad J. Domig ◽  
Dietmar Haltrich ◽  
...  

In the present study, we isolated and screened thirty strains of GABA (γ-aminobutyric acid)-producing lactic acid bacteria (LAB) from traditional Indonesian fermented foods. Two strains were able to convert monosodium glutamate (MSG) to GABA after 24 h of cultivation at 37 °C based on thin layer chromatography (TLC) screening. Proteomic identification and 16S rDNA sequencing using MALDI-TOF MS identified the strain as Lactobacillus plantarum designated as L. plantarum FNCC 260 and FNCC 343. The highest yield of GABA production obtained from the fermentation of L. plantarum FNCC 260 was 809.2 mg/L of culture medium after 60 h of cultivation. The supplementation of 0.6 mM pyridoxal 5’-phosphate (PLP) and 0.1 mM pyridoxine led to the increase in GABA production to 945.3 mg/L and 969.5 mg/L, respectively. The highest GABA production of 1226.5 mg/L of the culture medium was obtained with 100 mM initial concentration of MSG added in the cultivation medium. The open reading frame (ORF) of 1410 bp of the gadB gene from L. plantarum FNCC 260 encodes 469 amino acids with a calculated molecular mass of 53.57 kDa. The production of GABA via enzymatic conversion of monosodium glutamate (MSG) using purified recombinant glutamate decarboxylase (GAD) from L. plantarum FNCC 260 expressed in Escherichia coli was found to be more efficient (5-fold higher within 6 h) than the production obtained from fermentation. L. plantarum FNCC 260 could be of interest for the synthesis of GABA.


2022 ◽  
Vol 2022 ◽  
pp. 1-9
Author(s):  
Atefe Ghafurian Nasab ◽  
Sayed Ali Mortazavi ◽  
Farideh Tabatabaei Yazdi ◽  
Mahboobe Sarabi Jamab

In the present research, the production potential of gamma aminobutyric acid (GABA) using Lactobacillus brevis PML1 was investigated. In addition, the microorganism viability was examined in MAN, ROGOSA, and SHARPE (MRS) after undergoing high hydrostatic pressure at 100, 200, and 300 MPa for 5, 10, and 15 min. Response surface methodology (RSM) was applied to optimize the production conditions of GABA as well as the bacteria viability. Analysis of variance (ANOVA) indicated that both the independent variables (pressure and time) significantly influenced the dependent ones (GABA and bacteria viability) ( P < 0.05 ). The optimum extraction conditions to maximize the production of GABA included the pressure of 300 MPa and the time of 15 min. The amount of the compound was quantified using thin-layer chromatography (TLC) and spectrophotometry. For the process optimization, a central composite design (CCD) was created using Design Expert with 5 replications at the center point, whereby the highest content of GABA was obtained to be 397.73 ppm which was confirmed by high performance liquid chromatography (HPLC). Moreover, scanning electron microscopy (SEM) was utilized to observe the morphological changes in the microorganism. The results revealed that not only did have Lactobacillus brevis PML1 the potential for the production of GABA under conventional conditions (control sample) but also the content of this bioactive compound could be elevated by optimizing the production parameters.


2020 ◽  
Vol 15 (9) ◽  
pp. 1934578X2095328
Author(s):  
Xin Liu ◽  
Tian Deng ◽  
Xueqin Liu ◽  
Xiaohua Lai ◽  
Yanli Feng ◽  
...  

Oligosaccharides have important alimental and medical applications. Dextranase has been used to produce isomalto-oligosaccharides (IMOs). In this study, we isolated dextranase-producing bacteria from sugarcane-cultivated soil. Identification of the isolate based on its phenotypical, physiological, and biochemical characteristics, as well as 16S ribosomal deoxyribonucleic acid gene sequencing yielded Shewanella sp. strain GZ-7. The molecular weight of the dextranase produced by this strain was 100-135 kDa. The optimum temperature and pH for dextranase production were 40 °C and 7.5, respectively. The enzyme was found to be stable at the pH range of 6.0-8.0 and the temperature range of 20 °C-40 °C. Thin-layer chromatography and high-performance liquid chromatography of the enzymolysis products of the substrate confirmed the enzyme to be endodextranase. Under the optimal conditions, the ratio of IMOs could reach 91.8% of the hydrolyzate. The final products were found to efficiently scavenge the 2,2-diphenyl-1-picrylhydrazyl, hydroxyl, and superoxide anion radicals. In general, dextranase and hydrolyzates have high potential prospects for application in the future.


Author(s):  
Ida Bagus Agung Yogeswara ◽  
Suwapat Kittibunchakul ◽  
Endang Sutriswati Rahayu ◽  
Konrad J. Domig ◽  
Dietmar Haltrich ◽  
...  

In the present study, we isolated and screened thirty strains of GABA-producing lactic acid bacteria (LAB) from Indonesian traditional fermented foods. Two strains were able to convert monosodium glutamate (MSG) to GABA after 24 h of cultivation at 37oC based on thin layer chromatography (TLC) screening. 16S rDNA sequencing and proteomic identification using MALDI-TOF MS identified these two strains as Lactobacillus plantarum designated as L. plantarum FNCC 260 and L. plantarum FNCC 343. The highest yield of GABA production obtained from the fermentation of L. plantarum FNCC 260 was 809.2 mg/l of culture medium after 60 h of cultivation. Supplementation of 0.6 mM pyridoxal 5&rsquo;-phosphate (PLP) and 0.1 mM pyridoxine led to the increase in GABA production to 945.3 mg/l and 969.5 mg/l, respectively. The highest GABA production of 1226.5 mg/l of culture medium was obtained with 100 mM initial concentration of MSG added in the cultivation medium. The open reading frame (ORF) of 1410 bp of the gadB gene from L. plantarum FNCC 260 encodes 469 amino acids with a calculated molecular mass of 53.57 kDa. The production of GABA via enzymatic conversion of monosodium glutamate (MSG) using purified recombinant glutamate decarboxylase (GAD) from L. plantarum FNCC 260 expressed in Escherichia coli was found to be more efficient (5-fold higher within 6 h) than the production obtained from fermentation. L. plantarum FNCC 260 could be of interest for the synthesis of GABA.


2012 ◽  
Vol 586 ◽  
pp. 85-91
Author(s):  
Ying Guo Lü ◽  
Hui Zhang ◽  
Hui Yuan Yao

An enzymatic method for γ-aminobutyric acid (GABA) production was invested. With this method, rice bran was used as glutamate decarboxylase (GAD) source and exogenous monosodium glutamate(MSG) was used as substrate. We stimulated the rice bran GAD via regulating the temperature, pH, reaction time, buffer and adding PLP, Ca2+ and substrate. In the existence of PLP and Ca2+, the GABA content of rice bran had been improved by about 45 fold. The GABA production reached 2.3g/100g bran, and the Glu conversion reached 100%. As rice bran is a by-product in rice processing and a large quantity of rice bran is commercially available, our study illuminated a safe and efficient way to produce GABA and GABA enriched food.


2021 ◽  
Vol 63 (1) ◽  
pp. 42-48
Author(s):  
Dai Hung Ngo ◽  
◽  
Quoc Tuan Tran ◽  
Thi Nhat Hang Nguyen ◽  
Dai Nghiep Ngo ◽  
...  

Gamma-aminobutyric acid (GABA) is a potent bioactive component that widely exists in both plants and animals, has numerous health benefits. This study aimed to optimise the fermentation process conditions for the growth of Lactobacillus fermentum from rice bran extracts that have high potential to produce GABA. GABA content was assessed by thin-layer chromatography (TLC) method. In this study, fermenting conditions for medium production of GABA by L. fermentum from rice bran extracts were optimised. L. fermentum showed high potential for GABA-producing ability. Some factors influencing the GABA production such as carbon sources, nitrogen sources, mineral salt sources, substrate concentration of monosodium glutamate (MSG), pH, and the time of fermentation were investigated. When the L. fermentum is cultivated in the rice bran extracts medium supplemented with 1.5% lactose, 2% yeast extract, and 1% MSG with pH 6.0 in 48 h, this strain showed high GABA at a concentration of 736 mg/l.


2021 ◽  
Vol 9 (12) ◽  
pp. 2457
Author(s):  
Divakar Dahiya ◽  
Jemima V. Manuel ◽  
Poonam Singh Nigam

Gamma-aminobutyric acid (GABA) is an important chemical compound in the human brain. GABA acts as an inhibitory neurotransmitter by inducing hyperpolarization of cellular membranes. Usually, this pharmaceutically important compound is synthesized using a chemical process, but in this short overview we have only analysed microbial processes, which have been studied for the biosynthesis of this commercially important compound. The content of this article includes the following summarised information: the search for biological processes showed a number of lactic acid bacteria and certain species of fungi, which could be effectively used for the production of GABA. Strains found to possess GABA-producing pathways include Lactobacillus brevis CRL 1942, L. plantarum FNCC 260, Streptococcus salivarius subsp. thermophilus Y2, Bifidobacterium strains, Monascus spp., and Rhizopus spp. Each of these strains required specific growth conditions. However, several factors were common among these strains, such as the use of two main supplements in their fermentation medium—monosodium glutamate and pyridoxal phosphate—and maintaining an acidic pH. Optimization studies of GABA production were comprised of altering the media constituents, modifying growth conditions, types of cultivation system, and genetic manipulation. Some strains increased the production of GABA under anaerobic conditions. Genetic manipulation focused on silencing some genes or overexpression of gadB and gadC. The conclusion, based on the review of information available in published research, is that the targeted manipulation of selected microorganisms, as well as the culture conditions for an optimised bioprocess, should be adopted for an increased production of GABA to meet its increasing demand for food and pharmaceutical applications.


2021 ◽  
Vol 8 (1) ◽  
pp. 25-32
Author(s):  
Adhitya Naufal Pribadhi ◽  
Endang Kusdiyantini ◽  
Rejeki Siti Ferniah

Isolation and Characterization of Lactic Acid Bacteria from Fermented Food Cincalok as Producer of Gamma-Aminobutyric Acid Cincalok is a fermented food originating from West Kalimantan. This study aimed to obtain lactic acid bacterial isolates (LAB) capable of producing gamma-aminobutyric acid (GABA), to characterize the LAB isolates obtained, and to obtain GABA by the Thin Layer Chromatography (TLC) method. Bacterial growth and GABA production was carried out by adding 5% MSG and without MSG, and measured spectrophotometrically. In this study, 4 LAB bacterial isolates were obtained which were coded CIN-1, CIN-2, CIN-3, and CIN-4. GABA identification of all the LAB isolates using TLC Silica Gel 60 F254 with butanol: acetic acid: distilled water (5: 3: 2) as eluent yielded Rf 0.61 and Rf MSG 0.38. The highest growth was achieved by isolate CIN-3 with an absorbance of 1.488 (at 48 hour) in non-MSG medium, while the addition of 5% MSG resulted in an absorbance of 1.631 (at 42 hour). GABA production was achieved by isolate CIN-3 with 5% MSG treatment with a concentration of 201.472 mM and without MSG with a concentration of 171.195 mM. Cincalok merupakan pangan fermentasi yang berasal dari Kalimantan Barat. Penelitian ini bertujuan untuk mendapatkan isolat bakteri asam laktat (BAL) yang mampu menghasilkan gamma-aminobutyric acid (GABA), melakukan karakterisasi isolat BAL yang diperoleh dan dapat diperoleh GABA dengan metode Kromatografi Lapis Tipis (KLT). Penumbuhan bakteri dan produksi GABA dilakukan dengan penambahan MSG 5% dan tanpa MSG, dan diukur menggunakan spektrofotometer. Dalam penelitian ini diperoleh 4 isolat bakteri BAL yang diberi kode CIN-1, CIN-2, CIN-3, dan CIN-4. Identifikasi GABA dari semua isolat BAL tersebut menggunakan KLT Silica Gel 60 F254 dengan eluen butanol: asam asetat: aquades (5: 3: 2), menghasilkan Rf 0,61 dan Rf MSG 0,38. Pertumbuhan tertinggi terjadi pada isolat CIN-3 non MSG dengan absorbansi 1,488 (jam ke-48), sedangkan dengan penambahan MSG 5% menghasilkan absorbansi 1,631 (jam ke-42). Produksi GABA dicapai isolat CIN-3 dengan perlakuan MSG 5% dengan konsentrasi 201.472 mM dan tanpa MSG dengan konsentrasi 171,195 mM.


2019 ◽  
Vol 10 (5) ◽  
pp. 579-587 ◽  
Author(s):  
J.A. Valenzuela ◽  
A.B. Flórez ◽  
L. Vázquez ◽  
O.M. Vasek ◽  
B. Mayo

γ-Aminobutyric acid (GABA), an amino acid not used in protein synthesis, intervenes in several physiological functions and has both diuretic and calming effects in humans. Lactic acid bacteria (LAB) strains that produce GABA could be exploited for the manufacture of health-promoting GABA-enriched dairy products. In this study, 262 LAB strains isolated from traditional dairy products made from raw milk without starter cultures were screened for GABA production in culture media supplemented with 1% monosodium glutamate (MSG) using an enzymatic (GABase) method. About half of the strains (123) were found to be GABA producers. Of these, 24, among which were 16 Lactococcus lactis subsp. lactis and three Streptococcus thermophilus strains, produced >1 mM of GABA (range 1.01-2.81 mM) and were selected for further characterisation. GABA production was confirmed in most strains by culturing in 5 mM MSG followed by HPLC quantification. A majority of the strains were confirmed to be GABA producers by this method, although lower production levels were recorded. Using species-specific primers, the gene encoding glutamate decarboxylase (GAD) was PCR-amplified in all but one of the GABA producers analysed. Amplicons sequences were compared to one another and to those held in databases. Except for one Lactobacillus brevis strain, none of the 24 GABA producers investigated produced toxic biogenic amines, such as tyramine, histamine or cadaverine. They were therefore considered safe. Either alone, in mixtures, or in combination with industrial starter or adjunct cultures, these strains might be useful in the development of health-oriented dairy products.


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