• Title/Summary/Keyword: Thin Layer Chromatography(TLC)

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Biochemical Characterization of an Extracellular Xylanase from Aestuariibacter sp. PX-1 Newly Isolated from the Coastal Seawater of Jeju Island in Korea (대한민국 제주도 연안 해수에서 새롭게 분리한 Aestuariibacter sp. PX-1이 생산하는 자일라네이즈의 생화학적 특성)

  • Kim, Jong-Hee
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.215-222
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    • 2020
  • The marine microorganism PX-1, which can hydrolyze xylan, was isolated from coastal sea water of Jeju Island, Korea. Based on the 16S rRNA gene sequence and chemotaxonomy analysis, PX-1 was identified as a species of the genus Aestuariibacter and named Aestuariibacter sp PX-1. From the culture broth of PX-1, an extracellular xylanase was purified to homogeneity through ammonium sulfate precipitation and subsequent adsorption chromatography using insoluble xylan. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography estimated the molecular weight of the purified putative xylanase (XylA) as approximately 64 kDa. XylA showed xylanase activity toward beechwood xylan, with a maximum enzymatic activity at pH 6.0 and 45℃. Through thin-layer chromatographic analysis of the xylan hydrolysate produced by XylA, it was confirmed that XylA is an endo-type xylanase that decomposes xylan into xylose and xyloligosaccharides of various lengths. The Km and Vmax values of XylA for beechwood xylan were 27.78 mM and 78.13 μM/min, respectively.

Antioxidant activity of partially characterized polysaccharides from the edible mushroom Pleurotus djamor var. roseus

  • Raman, Jegadeesh;Sivakumar, Archana;Lakshmanan, Hariprasath;Raaman, Nanjian;Shin, Hyun-Jae
    • Journal of Mushroom
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    • v.19 no.3
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    • pp.140-149
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    • 2021
  • Mushroom-derived polysaccharides, which are the primary bioactive constituents, are beneficial for human health. Polysaccharides have immuno-modulation, antitumor, and antioxidant properties. Additionally, they have antiviral properties and protect against chronic radiation stress. In this study, high yield water-soluble polysaccharides were obtained from Pleurotus djamor var. roseus basidiocarps. The crude polysaccharide (CP) was extracted sequentially by hot water and ethanol precipitation. The yield of the brown CPs was 5.6% dw. Diethylaminoethyl cellulose and Sepharose-6B column chromatography of CPs generated several fractions. Total glucan content was determined in all the fractions. The F1 fraction displayed the highest sugar content and was considered as a purified polysaccharide (PP). The total glucan and β-glucan content in the four fractions ranged between 76.85-2.95% and 75.08-1.46%, respectively. The yield of the PPs was 300 mg, and it was obtained as a white powder. The PPs were characterized by Fourier-transform infrared spectroscopy (FTIR) and thin-layer chromatography. The FTIR spectral details confirmed the presence of a xylopentose polysaccharide. The antioxidant activity of the PPs was evaluated using in vitro 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay and superoxide radical scavenging assay. The PPs showed strong DPPH free radical and superoxide anion radical scavenging activities in a dose-dependent manner. Purified PPs free of phenolics, protein, and carbohydrates were mainly responsible for the radical scavenging activity. The data suggest the potential of PPs as natural antioxidants.

BEEF MEAT TRACEABILITY. CAN NIRS COULD HELP\ulcorner

  • Cozzolino, D.
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1246-1246
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    • 2001
  • The quality of meat is highly variable in many properties. This variability originates from both animal production and meat processing. At the pre-slaughter stage, animal factors such as breed, sex, age contribute to this variability. Environmental factors include feeding, rearing, transport and conditions just before slaughter (Hildrum et al., 1995). Meat can be presented in a variety of forms, each offering different opportunities for adulteration and contamination. This has imposed great pressure on the food manufacturing industry to guarantee the safety of meat. Tissue and muscle speciation of flesh foods, as well as speciation of animal derived by-products fed to all classes of domestic animals, are now perhaps the most important uncertainty which the food industry must resolve to allay consumer concern. Recently, there is a demand for rapid and low cost methods of direct quality measurements in both food and food ingredients (including high performance liquid chromatography (HPLC), thin layer chromatography (TLC), enzymatic and inmunological tests (e.g. ELISA test) and physical tests) to establish their authenticity and hence guarantee the quality of products manufactured for consumers (Holland et al., 1998). The use of Near Infrared Reflectance Spectroscopy (NIRS) for the rapid, precise and non-destructive analysis of a wide range of organic materials has been comprehensively documented (Osborne et at., 1993). Most of the established methods have involved the development of NIRS calibrations for the quantitative prediction of composition in meat (Ben-Gera and Norris, 1968; Lanza, 1983; Clark and Short, 1994). This was a rational strategy to pursue during the initial stages of its application, given the type of equipment available, the state of development of the emerging discipline of chemometrics and the overwhelming commercial interest in solving such problems (Downey, 1994). One of the advantages of NIRS technology is not only to assess chemical structures through the analysis of the molecular bonds in the near infrared spectrum, but also to build an optical model characteristic of the sample which behaves like the “finger print” of the sample. This opens the possibility of using spectra to determine complex attributes of organic structures, which are related to molecular chromophores, organoleptic scores and sensory characteristics (Hildrum et al., 1994, 1995; Park et al., 1998). In addition, the application of statistical packages like principal component or discriminant analysis provides the possibility to understand the optical properties of the sample and make a classification without the chemical information. The objectives of this present work were: (1) to examine two methods of sample presentation to the instrument (intact and minced) and (2) to explore the use of principal component analysis (PCA) and Soft Independent Modelling of class Analogy (SIMCA) to classify muscles by quality attributes. Seventy-eight (n: 78) beef muscles (m. longissimus dorsi) from Hereford breed of cattle were used. The samples were scanned in a NIRS monochromator instrument (NIR Systems 6500, Silver Spring, MD, USA) in reflectance mode (log 1/R). Both intact and minced presentation to the instrument were explored. Qualitative analysis of optical information through PCA and SIMCA analysis showed differences in muscles resulting from two different feeding systems.

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Composition Ratio Analysis of Transesterification Products of Olive Oil by Using Thin Layer Chromatography and Their Applicability to Cosmetics (올리브 오일의 에스터 교환반응 생성물의 TLC를 이용한 조성비 분석 및 화장품에의 응용가능성 평가)

  • Park, So Hyun;Shin, Hyuk Soo;Kim, A Rang;Jeong, Hyo Jin;Xuan, Song Hua;Hong, In Kee;Lee, Dae Bong;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.29 no.3
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    • pp.342-349
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    • 2018
  • In this study, the physicochemical properties, emulsifying capacity, moisture content and cytotoxicity of the composite material produced by transesterification reactions of the olive oil (olive oil esters) were investigated for cosmetic applications. Olive oil esters with short (S) and long (L) reaction times were studied. From the TLC-image analysis, composition ratios of the olive oil esters S were found to be 5.2, 24.1, 46.4, and 21.9% for mono-, di-, tri-glyceride, and fatty acid ethyl ester, respectively. Those of the olive oil esters L were 4.1, 24.7, 40.6, and 28.8% for mono-, di-, tri-glyceride, and fatty acid ethyl ester, respectively. The iodine value, acid value, saponification value, unsaponified matter, refractive index, and specific gravity were determined and purity tests were also carried out and normalized to establish standards and testing methods for using olive oil esters in cosmetics. To evaluate their emulsifying capacities, the O/W emulsion was prepared without surfactants and the formation of the emulsified particles were confirmed. After 5 days of applying the olive oil esters to human skin, the skin moisture retention was improved by 13.1% from the initial state. For the evaluation of toxicity on human skin cells, the olive oil esters showed 90% or more of the cell viability at $0.2-200{\mu}g/mL$. These results suggested that olive oil esters can be applied as natural/non-toxic ingredients to cosmetics industries.

The Screening of Aflatoxin Producing Fungi from Commercial Meju and Soy Bean Paste in Western Gyeongnam by Immunoassay (면역분석기법을 이용한 서부경남 시판 메주 및 된장에서의 Aflatoxin 생성균 검색)

  • 박정현;강성조;오상석;정덕화
    • Journal of Food Hygiene and Safety
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    • v.16 no.4
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    • pp.274-279
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    • 2001
  • Generally, non-aflatoxigenic fungi, such as Aspergillus oryzae, and Aspergillus are main microflora in Korean traditional fermented foods including Meju and soybean paste, but sometimes, Aspergillus flavus and Aspergillus parasiticus can be contaminated and accumulated aflatoxins during fermentation and storage. So the screening of aflatoxigenic strains in fermented traditional food is very important to improve the sanitary quality of those foods. In this work, we screened aflatoxin producing fungi from commercial Meju and soybean paste in Western Gyeongnam by immunoassay. Samples were randomly purchased from market of the commercial Meju(10 EA) and soybean paste(20 EA) in nine areas of Western Gyeongnam. Of the samples collected,24 strains and 22 strains of Aspergillus sp. were isolated from Meju and soybean paste, respectively. The isolated strains were cultured on SLS media at $25^{\circ}C$ for 15 days. The cultured broth were extracted with ethyl acetate and were analysed to determine aflatoxin B$_1$(AFB$_1$) by direct competitive ELISA(DC-ELISA). Six strains(25%) isolated from Meju, and 2 strains(9%) isolated from saybean paste, were confined as aflatoxin producing strains. The average range of aflatoxin productivity of isolates from Meju was 54.6 $\pm$ 38.7 ng/ml and that from soybean paste was 11.1 $\pm$ 8.6 ng/ml, respectively. Among them, isolated strain No. M-5-4 produced a high level of AFBl and showed 98.26 ng/ml of AFB$_1$. Every isolates were also re-confined their AFB$_1$productivity by thin layer chromatography(TLC). The TLC results also showed same trend as DC-ELISA results. As the above results, the screening of hazard mycotoxigenic fungi from traditional fermented foods should be necessary for the safety and the application of HACCP system in the food manufactory in Korea.

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Studies on the Determination Method of Natural Sweeteners in Foods - Licorice Extract and Erythritol (식품 중 감초추출물 및 에리스리톨 분석법에 관한 연구)

  • Hong Ki-Hyoung;Lee Tal-Soo;Jang Yaung-Mi;Park Sung-Kwan;Park Sung-Kug;Kwon Yong-Kwan;Jang Sun-Yaung;Han Ynun-Jeong;Won Hye-Jin;Hwang Hye-Shin;Kim Byung-Sub;Kim Eun-Jung;Kim Myung-Chul
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.258-266
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    • 2005
  • Licorice Extract and Erythritol, food additives used in korea, are widely used in foods as sweetener. Its application for use in food is regulated by the standard and specification for food additives but official analytical method far determination of these sweetener in food has not been established. Accordingly, we has been carried out to set up analytical method of the glycyrrhizic acid in several foods by the way of thin layer chromatography and high performance liquid chromatography glycyrrhizic acid is qualitative anaylsis technique consists of clean-up with a sep-pak $C_{18}$ cartridge, separation of the sweeteners by Silica gel 60 F254 TLC plate using 1-butanol:4Nammonia solution:ethanol (50:20:10) as mobile solvent. Also, the quantitative analysis for glycyrrhizic acid, was performed using Capcell prk $C_{18}$ column at wavelength 254nm and DW:Acetonitrile (62:38 (pH2.5)) as mobile phase. and we has been carried out to set up analytical method of the erythritol in several foods by the way of high performance liquid chromatography. erythritol is qualitative anaylsis technique consists of clean-up with a DW and hexane. The quantitative analysis for erythritol, was performed using Asahipak NH2P-50 column, Rl and DW:Acetonitrile (25:75) as mobile phase. The glycyrrhizic acid results determined as glycyrrhizic acid in 105 items were as follows; N.D$\∼$48.7ppm for 18 items in soy sauce, N.D$\∼$5.3ppm for 12 items in sauce, N.D$\∼$988.93ppm for 15 items in health food, N.D$\∼$180.7ppm for 26 items in beverages, N.D$\∼$2.6ppm for 8 items in alcoholic beverages repectively and ND for 63 items in the ethers. The erythritol results determined as erythritol in 52 items were as follows; N.D$\∼$155.6ppm for 13 items in gm, N.D$\∼$398.1ppm for 12 items in health foods repectively and ND for 45 items in the others.

Comparison of Carotenoid Pigments on Manchurian Trout, Brachymystax lenok and Masu Salmon, Oncorhynchus macrostomus in the Family Salmonidae (연어과에 속하는 열목어와 산천어의 Carotenoid 색소성분의 비교)

  • BAEK Sung-Han;HA Bong-Seuk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.2
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    • pp.278-287
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    • 1998
  • Carotenoids in integument of wild manchurian trout, Brachymystax lenok, and wild and cultured masu salmon Oncohynchus macrostomus, which are all the Korean native cold fresh water fish, were investigated by thin layer chromatography, column chromatography and HPLC. The total carotenoid contents of the wild manchurian trout were $3.72\;mg\%$ which is relatively higher compare to other species of salmonidae. The carotenoids were composed of $36.9\%$ zeaxanthin and $14.7\%$ $\beta-carotene$ as the major compounds, $7.8\%$ triol $7.3\%$ isocryptoxanthin, $5.7\%$ 4-hydroxy echinenone, $4.7\%$ lutein, $4.5\%$ salmoxanthin and $2.2\%$ astaxanthin as minor compounds, and other carotenoids such as canthaxanthin, tunaxanthin A, tunaxanthin B, tunaxanthin C, $\beta-cryptoxanthin$ and $\alpha-cryptoxanthin$ as minute carotenoids. Wild masu salmon contained more total carotenoids than cultured one and the contents were $0.82\;mg\%$ and $0.66\;mg\%$, respectively. The composition of the carotenoids from wild masu salmon were $20.7\%$ xeaxanthin, $17.0\%$ isocryptoxanthin and $15.8\%\;\beta-carotene$ as major compounds, and $6.2\%$ triol, $6.1\%$ 4-hydroxy echinenone, $6.1\%$ salmoxanthin, $5.9\%$ canthaxanthin, $5.8\%$ lutein, $4.9\%$ $\alpha-cryptoxanthin$ and $1.0\%$ astaxanthin as minor compounds. The composition of the carotenoids from cultured masu salmon were $19.7\%$ isocryptoxanthin, $18.0\%$ $\beta-carotene$ and $10.3\%$ zeaxanthin as the major compounds, and $8.9\%\;\beta-cryptoxanthin$, $8.5\%\;\alpha-cryptoxanthin$, $8.0\%$ lutein, $7.6\%$ canthaxanthin, $5.1\%$ triol and $2.0\%$ astaxanthin as minor carotenoids. Based on these data, wild masu salmon contained more zeaxanthin, salmoxanthin and 4-hydroxy echinenone while cultured masu salmon contained more $\alpha-cryptoxanthin$, indicating that carotenoid pigment of masu salmon depends on their living conditions. Unlike wild masu salmon, 4-hydroxy echinenone and salmoxanthin which are the characteristic carotenoids of salmons, were not found in the integument of cultured masu salmon. Unlike manchurian trout, both wild and cultured masu salmon did not contain tunaxanthin A, tunaxanthin B and tunaxanthin C.

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A Study on the Synthesis, Labeling and Its Biodistribution of Estradiol Derivatives (에스트라디올 유도체의 합성, 표지 및 체내동태에 관한 연구)

  • Kim, Sang-Wook;Yang, Seung-Dae;Suh, Yong-Sup;Chun, Kwon-Soo;Ahn, Soon-Hyuk;Lim, Soo-Jung;Choi, Chang-Woon;Lim, Sang-Moo;Kim, Young-Soon;Yu, Kook-Hyun
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.5
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    • pp.403-409
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    • 2000
  • Objectives: Due to the heterogeneous receptor distribution and changes of receptor status over time, the biochemical measurement of estrogen receptor status of biopsy specimens is not sufficient to diagnose breast cancer. As a result, I-123 labeled estradiols have been applied for the diagnosis. The purpose of this study was to develop a suitable radioligand for imaging estrogen receptor-positive human breast tumors. Methods: Among the various estradiol derivatives, $17{\alpha}-[^{123}I]$iodovinyl estradiol ($[^{123}I]$IVE) has been prepared from $17{\alpha}$-ethynyl estradiol. Labeling of $E-17{\alpha}-[^{123}I]$iodovinyl estradiol (E-$[^{123}I]$IVE) was carried out using peracetic acid with $[^{123}I]NaI\;and\;Z-[^{123}I]IVE$ labelling was archived using chloamine-T/HCl solution with $[^{123}I]$NaI. Labeling yield was determined by silica thin-layer chromatography (TLC) and radiochemical purity was measured by high performance liquid chromatography (HPLC). The biodistribution of E-$[^{123}I]$IVE was measured in immature female rats at 60 min, 120 min and 300 min after injection. Results: The labeling yield of two isomers was 92% and 94% ($E-[^{123}I]IVE\;and\;Z-[^{123}I]IVE$, respectively). The radiochemical purity was more than 98% after purification. The highest uptake was observed at 120 min in uterus (3.11% ID/g for E-$[^{123}I]$IVE). Conclusion: These results suggest the possibility of using E-$[^{123}I]$IVE as an imaging agent for the evaluation of the evaluation of the presence of estrogen receptor in patients with breast cancer.

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Identification and Quantification of Phytosterols in Maize Kernel and Cob (옥수수 종실 및 속대의 Phytosterol 동정과 함량 변이)

  • Kim, Sun-Lim;Kim, Mi-Jung;Jung, Gun-Ho;Lee, Yu-Young;Son, Beom-Young;Kim, Jung-Tae;Lee, Jin-Seok;Bae, Hwan-Hee;Go, Young-Sam;Kim, Sang-Gon;Baek, Seong-Bum
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.63 no.2
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    • pp.131-139
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    • 2018
  • Unsaponifiables in the kernel and the cob of 7 maize varieties were analyzed by using thin-layer chromatography (TLC) and gas chromatography (GC) for the identification of phytosterols and their concentrations. The unsaponifiables of the kernel were clearly separated into band I (campesterol, stigmasterol, and ${\beta}$-sitosterol), band II (${\Delta}^5$-avenasterol), band III (${\Delta}^7$- stigmastenol), and band IV (${\Delta}^7$-avenasterol). In the cob, on the other hand, three or more bands were separated in addition to bands. The GC analysis of unsaponifiables showed good separation of campesterol, stigmasterol and ${\beta}$-sitosterol, but the mixture of ${\Delta}^7$-avenasterol (retention time[RT] 22.846), ${\Delta}^7$-stigmastenol (RT 22.852), and ${\Delta}^5$-avenasterol (RT 22.862) showed poor separation. Phytosterol content of the maize kernel was 635.9 mg/100 g, and that of the cob was 273.0 mg/100 g, respectively. The phytosterol content of the kernel was 2.4-fold higher than that of the cob. The phytosterol content of the kernel was higher in the order ${\beta}$sitosterol 80.05% > campesterol 10.5% > stigmasterol 9.46%, but that of the cob was higher in the order ${\beta}$-sitosterol 59.43% > stigmasterol 31.72% > campesterol 10.98%. Based on these results, it appears that the phytosterols of the maize kernel are synthesized in the maize cob and are transferred to the kernel, because the precursors (${\Delta}^7$-avenasterol, ${\Delta}^7$-stigmastenol, and ${\Delta}^5$-avenasterol) of major phytosterols were detected in maize cobs.

Production of γ-amino Butyric Acid by Lactic Acid Bacteria in Skim Milk (탈지방우유에서 가바생성 유산균 배양을 통한 가바생성 연구)

  • Cha, Jin Young;Kim, Young Rok;Beck, Bo Ram;Park, Ji Hun;Hwang, Cher Won;Do, Hyung Ki
    • Journal of Life Science
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    • v.28 no.2
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    • pp.223-228
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    • 2018
  • Lactic acid bacteria were isolated from a variety of fermented seafoods and sea creatures from the East Sea Rim, Korea and were screened for ${\gamma}-amino$ butyric acid-producing (GABA) activity. Through a 16S rRNA sequence analysis, the bacteria of interest, which were GABA-positive on the thin-layer chromatography analysis, were recognized as three isolates of Lactobacillus (Lb.) brevis and one isolate of Lactococcus (Lc.) lactis. Lb. brevis FSFL0004 and FSFL0005 were isolated from fermented anglerfish and Lb. brevis FSFL0036 was derived from salted cutlass fish. The Lc. lactis strain FGL0007 was isolated from the gut of a brown sole flounder. According to HPLC analysis, the GABA contents produced by FSFL0004, FSFL0005, FSFL0036 and FGL0007 were equivalent to $10,754.37{\mu}g/ml$, $13,082.79{\mu}g/ml$, $12,290.19{\mu}g/ml$, and $45.07{\mu}g/ml$ respectively in 1% monosodium glutamate-supplemented methionyl-tRNA synthetase (MRS) broth. The four strains were inoculated in skim milk with 1% monosodium glutamate to commercialize the strains as starter cultures for GABA-enriched dairy products, and TLC results displayed the production of ${\gamma}-amino$ butyric acid by all four strains in the adaptation media. Lc. lactis FGL0007 demonstrated the greatest GABA production ($431.42{\mu}g/ml$) by HPLC analysis. The GABA production by lactic acid bacteria strains in the skim milk demonstrated in the present study may be helpful for the production of GABA-enriched dairy products.