Bakteri Asam Laktat (BAL) adalah kelompok mikroorganisme General Recognized as Safe (GRAS) yang berperan penting dalam menghasilkan senyawa fungsional, termasuk Gamma-aminobutyric acid (GABA). GABA merupakan asam amino non-protein yang memiliki fungsi sebagai penghambat neurotransmitter utama dalam sistem saraf pusat juga berperan sebagai antioksidan. Penelitian ini bertujuan untuk mengetahui kemampuan BAL dari produk fermentasi Chao khas daerah Pangkep dalam menghasilkan senyawa GABA sebagai antioksidan menggunakan metode Thin Layer Chromatography (TLC). Berdasarkan penelitian yang dilakukan diperoleh nilai Retention factor (Rf) isolat BAL C2 tanpa MSG yaitu 0,82, C2 MSG 1% 0,84, C10 tanpa MSG 0,8 dan 0,64, C10 MSG 1% 0,86 dan 0,68. Dengan aktivitas antioksidan berdasarkan nilai IC50 C2 tanpa MSG 940,11 ppm, C2 MSG 1% 1124,97 ppm, C10 tanpa MSG 884,73 ppm, dan C10 MSG 1% 939,17 ppm. Nilai Rf isolat C2 dan C10 dengan penambahan MSG 1% lebih menghampiri nilai Rf larutan standar yaitu 0,88 dan aktivitas antioksidan isolat C2 dan C10 tergolong sangat lemah dikarenakan nilai IC50 nya >200 ppm. Ini dapat menjadi dasar optimalisasi produksi GABA oleh BAL dari produk fermentasi lokal seperti Chao untuk pengembangan probiotik atau suplemen fungsional yang bermanfaat untuk kesehatan. Kata kunci: Chao, Bakteri Asam Laktat, Gamma-Aminobutyric Acid, Antioksidan Lactic Acid Bacteria (LAB) are a group of microorganisms generally recognized as safe (GRAS) that play a vital role in producing functional compounds, including Gamma-aminobutyric acid (GABA). GABA is a non-protein amino acid that functions as a major inhibitory neurotransmitter in the central nervous system and also acts as an antioxidant. This study aimed to determine the ability of LAB from a local fermented product, Pangkep Chao, to produce GABA as an antioxidant using Thin Layer Chromatography (TLC). Based on the research, the retention factor (Rf) values for LAB isolates were as follows: C2 without MSG was 0.82, C2 with 1% MSG was 0.84, C10 without MSG was 0.8 and 0.64, and C10 with 1% MSG was 0.86 and 0.68. The antioxidant activity, based on the IC50 value, was 940.11 ppm for C2 without MSG, 1124.97 ppm for C2 with 1% MSG, 884.73 ppm for C10 without MSG, and 939.17 ppm for C10 with 1% MSG. The Rf values of the C2 and C10 isolates with 1% MSG were closer to the standard solution's Rf value of 0.88, but the antioxidant activity of the C2 and C10 isolates was considered very weak because their IC50 values >200 ppm. This research can serve as a basis for optimizing GABA production by LAB from local fermented products like Chao for the development of beneficial probiotics or functional supplements
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