Biji kefir air berupa matrik gel polisakarida, bening seperti kristal, didalamnya terdapat simbiosis kompleks antara bakteri asam laktat dan khamir. Penelitian ini bertujuan mendapatkan konsentrasi optimal biji kefir air sebagai pengembang roti kukus. Roti kukus dibuat dengan tiga konsentrasi biji kefir air: 75, 100, dan 125% berdasarkan 100% tepung, kemudian dianalisis sifat mutu fisik dan sensorinya. Roti kukus dengan pengembang biji kefir air terbaik dibandingkan sifat fisik dan kadar seratnya dengan yang diolah menggunakan khamir (standar). Hasil penelitian menunjukkan bahwa roti kukus yang dibuat dengan biji kefir air 100% memiliki mutu fisik (volume spesifik, struktur remah) lebih baik daripada yang diproses dengan biji kefir air 75% dan 125%. Mutu sensori roti kukus yang dibuat dengan 100% dan 125% biji kefir air (kenampakan, kehalusan permukaan, pori, tekstur, kelengketan, aroma) lebih tinggi dibandingkan yang diolah dengan biji kefir air 75%. Penggunaan biji kefir air 100% ditentukan sebagai konsentrasi bahan pengembang optimal pada pembuatan roti kukus. Roti kukus yang dibuat dengan biji kefir air 100% memiliki volume spesifik dan struktur remah tidak beda nyata dengan roti kukus standar, sedangkan kandungan serat pangannya lebih tinggi. Kandungan serat pangan berturut-turut roti kukus dengan biji kefir 100% adalah 18.5% (bk) dan roti kukus standar 12.5% (bk). Dapat disimpulkan bahwa konsentrasi biji kefir air optimal sebagai bahan pengembang roti kukus adalah 100% berdasarkan 100% tepung.Kata kunci: bakteri asam laktat (BAL), biji kefir air, khamir, roti kukus. APLICATION OF WATER KEFIR GRAINS AS LEAVENING AGENT OF STEAMED BREAD ABSTRACTWater kefir grain is a complex symbiosis between lactic acid bacteria and yeast in transparent gel matrix crystal-like. The objective of this study was to get optimum concentration of water kefir grains as leavening agent of steamed bread. Steamed bread was prepared with three concentrations of water kefir grains: 75, 100, and 125% based on 100% flour, then their physical and sensory properties were analyzed. The chosen steamed bread using water kefir grains was then compared its physical properties and fiber content to the standard steamed bread using yeast. Results showed that steamed bread made with 100% of water kefir grains had better physical properties (specific volume, crumb structure) than that of 75% and 125%. The sensory properties (appearance, surface smoothness, crumb structure, tenderness, adhesiveness, aroma) of steamed bread made with 100% and 125% of water kefir grains were superior than that of 75%. The use of 100% water kefir was chosen as the optimum concentration for leavening agent of steamed bread. Steamed bread using 100% water kefir had no differences on specific volume and crumb structure compared to those of standard steamed bread, whereas its fiber content was significantly higher. Fiber content of steamed bread using 100% water kefir grains and steamed bread using yeast was 18.51% (db) and 12.51% (db), respectively. It can be concluded that optimum concentration of water kefir grains used as leavening agent for steamed bread making was 100% based on 100% of flour.