Arabitol has several applications in food and pharmaceutical industries as a natural sweetener, dental caries inhibitor, and texturing agent. Newly isolated yeast strains from seawater, sugarcane plantation soil samples, and Zygosaccharomyces rouxii 2635 from MTCC were tested for arabitol production. The yield of arabitol was found to be higher in seawater isolate (24.6 g L −1 ) compared to two soil isolates (22.5 g L −1 ) and Z. rouxii (19.4 g L −1 ). Based on ITS 26S rDNA sequence analysis, the seawater isolate was identified as Pichia manchurica. In the present study, the effect of different substrates, trace elements, nitrogen sources, pH, and temperature on arabitol production was examined. Three different carbon sources viz. glucose, arabinose, and galactose were studied. Glucose was determined to be the best substrate for arabitol production (27.6 g L −1 ) followed by arabinose (13.7 g L −1 ) and galactose (7.7 g L −1 ). Maximum production of arabitol was observed at pH 6.0 (34.7 g L −1 ). In addition, arabitol production was high (35.7 g L −1 ) at temperature of 30 °C. Among the different concentrations of ammonium sulfate tested (3, 4.5, 6, 7.5, and 9 g L −1 ) concentration of 6 g L −1 resulted in higher arabitol Individual metal ions had no effect on arabitol production by this strain as compared to control. Results obtained in this study identify ways for improved arabitol production with natural isolates using microbial processes. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim