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.