The aim of this study was to evaluate the natural producer Lactobacillus diolivorans as potential production organism of 1,3-propanediol from glycerol. Different cultivation parameters, such as oxygen supply, feeding-strategy, or medium composition have been tested in batch and fed-batch cultivations. The 1,3-propanediol concentration obtained in batch cultivations was 41.7 g/l. This could be increased to 73.7 g/l in a fed-batch co-feeding glucose and glycerol with a molar ratio of 0.1. Yeast extract as part of the MRS cultivation medium could be replaced by nicotinic acid and riboflavin. Furthermore, the addition of Vitamin B-12 to the culture medium increased production by 15% to a final titer of 84.5 g/l 1,3-propanediol. (C) 2012 Elsevier Ltd. All rights reserved.