Abstract: The production of rifamycins B and SV by Amycolatopsis mediterranei using immobilization technique was investigated. Alginate beads proved to be an alternative method for the production of rifamycin antibiotic as it has many advantage over using free cells such as it enables the operation at higher dilution rates without the danger of wash out, ease of handling and facilitates recycling or reverse of using microorganism. Different hardening agents were used to improve the stability of the beads and decrease cells escapement. Gum Arabic and gellan gum were used and the rate of rifamycin production increased by using gellan gum as hardening agent compared to pure alginate beads. The rifamycins B and SV increased from 685 to 810 and from 547.5 to 702 mg L-1, respectively by using gellan-gum modified beads compared to the non-treated beads. In repeated batch production, the beads were stable and no breaking of the gel beads was observed throughout the experiment time. The total amount of rifamycin B and SV during the five repeated batches (each of 144 h) reached 3360 and 2309 mg L-1, respectively. On reducing the batch time to only 72 h, the antibiotic production was not influenced and the total concentration of rifamycins B and SV for five batches was 3785 and 2393 mg L-1, respectively.