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Mutants of with high erythritol production were generated through an atmospheric and room temperature plasma (ARTP) mutation system. Among these mutants, M53 exhibited the highest erythritol yield. In a batch culture, M53 produced 64.8 g/L erythritol from 100 g/L glycerol. The yields of byproducts (e.g. mannitol, arabitol, and α-ketoglutaric acid) were low, and the mechanisms underlying these changes were examined by measuring enzyme activities in the pentose phosphate pathway. Up to 145.2 g/L erythritol was produced by M53 from 200 g/L of glycerol, and erythritol accumulation was promoted by 3.7 mg/L of Cu, 10.15 mg/L of Mn, and 30.37 g/L of NaCl. Fed-batch cultivation of M53 in a 5-L fermentor produced 169.3 g/L erythritol with low levels of byproducts within 168 h. This finding confirmed the potential of M53 as an erythritol producer on a commercial scale.