Bryant made a pull-up jumper from the free-throw line with 7:54 remaining in the second quarter against Sacramento to eclipse Chamberlain's mark of 31,419 points. He had tied Chamberlain 22 seconds earlier with a layup.
Bryant entered the game four points behind Chamberlain.
He finished the game with 19 points and 14 assists in a 103-98 Lakers victory.
Kareem Abdul-Jabbar tops the NBA's career scoring list with 38,387 points, Karl Malone is second with 36,928 and Michael Jordan is third with 32,292.
The 34-year-old Bryant is in his 17th season — all with the Lakers.