Beatie, 38, revealed the break-up from his wife Nancy during a taping of the syndicated CBS show ‘The Doctors’.
He also admitted during the interview that he had undergone his final gender reassignment surgery from female to male.
‘Like all marriages, we have our ups and downs, and we’re going through a rough patch right now.
‘At the moment, we’re separated.’
Mr Beatie was born as a woman, Tracy Lagondino, in Hawaii in 1974, but says he always felt like he wanted to be a man.
When he was in his twenties he began having testosterone injections, giving him facial hair, a lower voice and altering his sexual organs.
In 2002 he had a mastectomy and legally became a man – but he chose to keep his vagina, uterus and other female sexual organs so the couple could have children, as his wife had had a hysterectomy.
He posed for an infamous picture in 2007 which displayed him as heavily pregnant but with facial hair.