Guatemalan President Jimmy Morales and Israeli Prime Minister Benjamin Netanyahu cut a blue-and-white ribbon as the official opening of the embassy.
Guatemala on Wednesday celebrated the opening of its new embassy in Jerusalem, following suit after the USA move that sparked violent protests along the Israeli-Palestinian border.
Guatemala on Wednesday opened its embassy in Jerusalem, becoming the second country to do so.
The US and Guatemalan moves break with decades of global consensus.
The embassy's opening was a fulfillment of one of Trump's 2016 campaign promises, but it was met with large protests from Palestinians, who see East Jerusalem as the future capital of a Palestinian state.
Israel's violent reaction to the protesters drew global criticism and mounting calls for an investigation.
This comes two-days after the United States (US) inaugurated its embassy in Jerusalem.
Netanyahu also promised to visit Guatemala in his next trip to Latin America, according to the several media reports.
But on Tuesday, senior United Nations officials condemned the killings as an "outrageous human rights violation" and said it appeared that protesters approaching the fence were liable to be shot and killed by Israeli forces without warning.
"You were always among the first", he said. "It seems that anyone is liable to be shot dead".
On Monday, Turkey recalled its ambassadors to the USA and Israel in reaction to the embassy move.