Regulation of hyaluronic acid molecular weight and titer by temperature in engineered Bacillus subtilis

Hyaluronic acid (HA) is a biopolymer used in several industries. There is increasing global demand. HA is normally produced on a large scale using attenuated strains of group C streptococci that are pathogenic and fastidious. Accordingly, it is of interest to use a "generally recognized as safe" (GRAS) organism such as Bacillus subtilis for HA production. Here, we report an engineered B. subtilis strain named WmB that produces different molecular weights (MW) and titers of HA at different temperatures. The faster the bacteria grew, the lower the MW of HA produced and the higher the titer. The MW of HA obtained ranged from 6.937 MDa at 47 °C to 0.392 MDa at 32 °C. At 32 °C, the HA titer reached 3.65 ± 0.13 g/L. We have engineered a strain that can produce high-molecular-weight and medium-molecular-weight HA at different growth temperatures. This GRAS B. subtilis strain can be applied in industry and provides a new strategy for production of HA with different molecular weights.