Bio-based 2,3-butanediol (2,3-BDO) production is still limited to the lab scale, although its advantages over the chemical route include optically pure isomer production and lower energy demand. Few previous studies have investigated bio-based 2,3-BDO scale-up using well-defined criteria and safe microorganisms. This study aimed to produce 2,3-BDO by a newly Paenibacillus peoriae NRRL BD-62 in a pilot bioreactor with 70 L, keeping the oxygen transfer rate (OTR) constant and below 1.68 mmolO2/L/h to ensure the microaerobic conditions. A surface aeration system was adopted, and an energy dissipation of 270.5 W/m3 and an impeller tip speed of 2.04 m/s were required. A 2,3-BDO productivity and yield of 0.90 g/L/h and 0.40 g/g, comparable to unsafe or genetically engineered microbial producers, were observed in the batch assay, respectively. No acetoin accumulation and a levo-2,3-BDO optical purity of about 98 % were achieved in 26 h. A delay in pulse feeding resulted in an ethanol coproduction (1:1 ratio) in the fed-batch assay, although the global productivity and yield were similar to the batch assay. To the best of our knowledge, this is the first time that 2,3-BDO has been produced by wild-type and safe P. peoriae strain in a pilot bioreactor keeping OTR constant.