Obama spoke alongside Prime Minister Benjamin Netanyahu after the two met privately at the White House on Wednesday.
"I did want to specifically take some time out to speak to the people of Israel and to the region about the senseless slaughter that took place near Hebron yesterday," Obama said.
"There are going to be extremists and rejectionists who, rather than seeking peace, are going to be seeking destruction, and the tragedy that we saw yesterday, where people were gunned down on the street by terrorists who are purposely trying to undermine these talks, is an example of what we're up against," Obama continued.
Obama expressed condolences from the American people to the families of the terror victims, and said that the U.S. is unwavering in its support of Israel's security.
"The United States is going to be unwavering in its support of Israel's security, and we are going to push back against these kinds of terrorist activities," said the president.
Obama also sent a message to Hamas, which claimed responsibility for the attack.
"This is not going to stop us from not only ensuring a secure Israel, but also securing a longer-lasting peace in which people throughout the region can take a different course," said Obama.
The U.S. president also said he appreciates the fact that Netanyahu remain committed to peace, despite Tuesday's attack.
Netanyahu applauded Obama's condemnation of Tuesday's killing, and said they were carried out by people "who do not respect human life and who trample human rights into the dust and butcher everything they oppose."
The U.S. is hosting Netanyahu and Palestinian President Mahmoud Abbas for a new round of Middle East peace talks, due to start Thursday. Obama is scheduled to meet Abbas later Wednesday.
Netanyahu earlier on Wednesday urged settlers in the West Bank to exhibit restraint and respect the rule of law in Israel in the wake of a deadly shooting attack near Hebron.
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