Marshall County Sheriff Kevin Byars said the explosion at the Silver Trail Distillery occurred around 10:45 a.m. on Friday. Two workers, 40-year-old Jay Rogers and 26-year-old Kyle Rogers, suffered severe burns and were flown to Vanderbilt University Medical Center in Nashville.
Byars said fire officials believe a still at the plant malfunctioned, causing the blast.
The distillery produces "LBL Moonshine," which is named for the nearby Land Between the Lakes National Recreation Area.
Fire officials extinguished the blaze, but the building is a total loss.
Distillery founder R. Spencer Balentine opened Silver Trail in 2011. The company's website says that Balentine's father was a local moonshine runner in the 1950s.