Thapelo Makutle was attacked at his place of work in the John Taolo Gaetsewe district on Friday, MambaOnline quoted the NGO Legbo Northern Cape as saying.
Makutle apparently got into an argument with two heterosexual men over his sexuality, and the attackers allegedly followed him to the room where he was staying.
He was found in the room early on Saturday morning with a wound to the neck, police say.
Makutle, who identified as both gay and transgender, was a volunteer for Legbo Northern Cape which provides support for LGBTI persons in rural communities.
He was recently crowned Miss Gay Kuruman and took part in the Kimberley Out In Africa Gay and Lesbian Festival.
"It's so sad. I can't describe the pain that we are feeling right now," Shaine Griqua, the Legbo Northern Cape director, told Mambaonline. "We have lost a young, talented, gay man who was open about who he was. The last few days have been like a dark cloud."
Griqua said that hate crimes were on the rise in the province. He said that a lesbian was recently attacked at a bar but chose not to press charges as she didn’t want to involve her family.
Cosatu in the Northern Cape also condemned the killing and urged the provincial government to call a summit on hate crime.
Makutle’s funeral will be held on Saturday in Kuruman.
*This report earlier said that the victim was beheaded, but police say that was not the case. They will only say that he had a neck wound.