"Your honor, I am an innocent man," Dr Conrad Murray told Superior Court Judge Michael Pastor in a soft voice. "I definitely plead not guilty."
Lawyers for Murray, who is accused of giving the "King of Pop" a lethal dose of the powerful anesthetic propofol and other sedative said they were ready to go to trial - which is highly likely to be televised.
"Dr. Murray is looking forward to finally telling his side of the story," defense attorney Ed Chernoff said outside court.
"He has been waiting 22 months for his opportunity to do this. It's the first chance we have to force the issue."
Murray will not seek a plea bargain but if convicted he could face a maximum for years in prison.
His behavior in the run up to the superstar's death has been well documented in testimony from household staff as well as e-mails and Murray's own police interview.
All will be used in evidence.
He told officers that he gave Jackson a low dose of propofol - which Jackson called: "his milk" - after spending 10 hours trying to get him to sleep using other drugs.
When the star appeared to doze off, Murray said he left the room for two minutes to go to the bathroom then returned to find Jackson not breathing.
He delayed calling 911 for between 25 minutes and an hour while he tried to revive him, testimony showed.
A coroner testified that Jackson, 50, died of a propofol overdose in combination with other drugs on June 25, 2009. His death was classified as a homicide.