In this study, after screening of 9 fungal strains for their ability to produce calcium malate, it was found that Penicillium sderotiorum K302 among them could produce high-level of calcium malate. Under the optimal conditions, the titer of calcium malate in the supernatant was 88.6 g/l at flask level. During 10-l fermentation, the titer of 92.0 g/l, the yield of 0.88 g/g of glucose and the productivity of 1.23 g/l/h were reached within 72 h of the fermentation, demonstrating that the titer, yield and productivity of calcium malate by this strain were very high and the fermentation period was very short. After analysis of the partially purified product with HPLC, it was found that the main product was calcium malate. The results showed that P. sclerotiorum K302 obtained in this study was suitable for developing a novel one-step fermentation process for calcium malate production from glucose on large scale. (C) 2013 Elsevier Ltd. All rights reserved.