The aim of this work was to study the random mutagenesis of Clostridium butyricum strain. A high 1,3-PD tolerant mutant strain, designated as C. butyricum YP855, was developed from the wild strain C. butyricum XYB11, using combined chemical (NTG, N-methyl-N'-nitro-N-nitrosoguanidine,) and plasma-based mutagenesis (ARTP, atmospheric and room temperature plasma). The YP855 showed a maximum tolerance of 85 g/L to 1,3-PD (up to 30.8% increase) when compared with the tolerance exhibited by the wild strain. Under the optimum conditions as established by the response surface methodology (RSM), the mutant strain produced 37.20 g/L of 1,3-PD, which is 29.48% higher than the concentration obtained from the wild strain (28.73 g/L). This research would offer information for further development of the biosynthesis of 1,3-PD by the mutant strain of C. butyricum.