Ronaldo had been tied on 56 with Madrid legend Hugo Sanchez since scoring from the spot in Madrid's 2-1 defeat to Sevilla on January 15.
But now he has the record all to himself having slammed home a penalty on Sunday after a Toni Kroos shot was controversially adjudged to have struck the arm of Bruno.
Ronaldo's successful spot-kick brought the match back to 2-2 after Madrid had earlier fallen two goals behind.
57 - Cristiano Ronaldo is now the player with most penalties scored in La Liga history, surpassing Hugo Sanchez (56). Punisher. pic.twitter.com/aWjmMezadO— OptaJose (@OptaJose) February 26, 2017
Barcelona superstar Lionel Messi lags some way behind Ronaldo in fifth with 43, also trailing Luboslav Penev (44) and Ronald Koeman (45).
Ronaldo's 57th spot-kick is his fifth penalty goal in LaLiga this season and takes his overall league tally for the campaign to 16.