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The objective of this study was to characterize the durian lay juices supplemented by Lactic Acid Bacteria (LAB) culture during cold storage. The LAB cultures consisted of Lactobacillus plantarum FNCC-0265, Lactobacillus acidophillus FNCC-0051 and combination of these LABs. After being inoculated with a 24 h – old culture starters, durian lay juice was incubated at 37 °C for 2 hours and kept at 4 °C for 0, 1, 2, 3 and 4 weeks. Changes in pH, total lactic acid, and sugar content, viable cell counts of LAB, total soluble solid, viability of LAB, and sensory characteristic during fermentation were measured. The results revealed that pH decreased from 4.79 to 4.20; sugar content reduced from 4.52 to 0.75 %; total soluble solid reduced from 8.93 to 7.97 °Bx, and sensory score reduced from 3.71 (like) to 3.40 (moderate like); while the total lactic acid increased from 0.33 to 0.69 %; viable cell counts of LAB increased from 8.86 to 9.14 log CFU/ml; and % viability of LAB decreased from 100 to 95.90, during storage. Combination of Lactobacillus plantarum FNCC-0265 and Lactobacillus acidophillus FNCC-0051 cultures at two weeks cold storage period was the best treatment, based on the sensory and viable cell counts of LAB. The characteristics of this durian lay juice probiotic were pH 4.46, total lactic acid 0.59 %, sugar content 2.13 %, viable cell counts of LAB 9.33 log CFU/mL, total soluble solid 9.60 °Bx, and carbohydrate 45,05 %. This durian lay juice contained oxalate (107.56 mg/L), malic (527.895 mg/L), lactic (4381.49 mg/L) and citric (351.245 mg/L) acid. ABSTRAK Tujuan penelitian ini adalah mengkaji karakteristik minuman laktat sari durian lay yang disuplementasi dengan kultur bakteri asam laktat (BAL) selama penyimpanan pada suhu rendah. Kultur BAL terdiri dari Lactobacillus plantarum FNCC-0265, Lactobacillus acidophillus FNCC-0051, dan campuran keduanya. Sari buah durian lay diinokulasi dengan kultur berumur 24 jam lalu diinkubasi pada suhu 37 °C selama 2 jam dan kemudian disimpan selama 0, 1, 2, 3, dan 4 minggu. Perubahan pH, total asam, gula, total sel BAL. Total padatan terlarut, viabilitas BAL, dan sensori selama penyimpanan dingin diukur. Hasil penelitian menunjukkan bahwa terjadi perubahan pH dari 4,79 ke 4,20; kandungan gula dari 4,52 ke 0,75%; total padatan terlarut dari 8,93 ke 7.97 °Bx, viabilitas BAL dari 100 % menjadi 95,90 %, dan skor sensori dari 3,71 suka ke 3,40 (agak suka); sementara pada total asam terjadi peningkatan dari 0,33 ke 0,69 %; dan sel hidup BAL meningkat dari 8,86 menjadi 9,14 log CFU/mL. Berdasarkan parameter sensori, perlakuan kultur campuran Lactobacillus plantarum FNCC-0265 dan Lactobacillus acidophillus FNCC-0051 yang disimpan selama 2 minggu merupakan perlakuan terbaik. Sari buah durian lay ini mempunyai karakteristik sebagai berikut: pH 4,45, total asam laktat 0,83 %, gula 2,20%, sel hidup BAL 9,33 log CFU/mL, total padatan terlarut 9,16 °Bx, dan karbohidrat 35,04 %. Asam organik yang terkandung pada jus durian lay ini adalah asam oksalat (108 mg/L), malat (528 mg/L), laktat (4382 mg/L) dan sitrat (351 mg/L).