The former world No 1 fell victim to one of the greatest shocks in Grand Slam history as Istomin condemned the Serb to his earliest exit at a major in nearly a decade 7-6 (10-8) 5-7 2-6 7-6 (7-5) 6-4 on Rod Laver Arena.
After winning four slams in a row between 2015 and 2016, Djokovic has now gone three without adding to his 12 major triumphs, raising more questions about his ability to stay at the top after dominating tennis for the last six years.
Djokovic denied there was any hangover lingering from his breakthrough French Open victory last June, and the underwhelming second half of last season which ensued
Asked if his long-awaited victory at Roland Garros had taken the edge off his hunger, Djokovic said: "I don't know. I mean, it's not a time now to go so deep into it.
"I didn't reflect on that at all at this stage. I started a new season, a new year, as everybody else did. I forgot about it, in a way. It's not affecting me."
Djokovic, who began the year by beating Andy Murray at the Qatar Open earlier this month, had lost to a player ranked outside the top 100 only once since 2010.
Defeat to Istomin, however, ranked 117th in the world, halts his revival and raises questions about his drive to stay at the top.
"I started the season very well. Again, it's a tennis match," Djokovic said. "On any given day, you can lose. I mean, nothing is impossible.
"There is over a hundred players playing in the main draw. I guess the quality of tennis keeps rising each year. Everybody becomes more professional. I guess they improve.
"They get better on the court. What can I do? I did try my best till the last shot, but it didn't work."
As the match slipped beyond Djokovic's control, he could conjure little emotion to rally in the fifth set but he denied that his ruthless, steely attitude he once depended on were lacking in the stunning upset.
"There was intensity, of course," Djokovic said. "We played four-and-a-half hours.
"It's just one of these days when you don't feel that great on the court, don't have much rhythm, and the player you're playing against is feeling the ball very well. That's sport.
"Of course, I'm not used to losing in the Australian Open second round. I've always played so well. Through the last 10 years, I've won six titles here.
"This court has been so nice to me. I enjoyed it very much. Of course, it's disappointing. But at the end of the day I have to accept it."
Djokovic's former coach Boris Becker told Eurosport: "I'm really shocked that Novak lost.
"I thought he was playing much too defensive and never really took the initiative and never really fought for it.
"I absolutely never expected him to have trouble with Denis Istomin. No disrespect but Novak is a six-time Australian Open champion.
"But I could tell in the first set he was way too defensive and way too passive. The first set in a three-out-of-five match is always very important.
"In the fourth set again he was a bit lethargic and not playing at full power.
"Then once you're in the fifth set anything can happen and credit to Denis, he kept his composure, he kept his nerve, he went a break up and kept serving well and hitting the lines so it's well deserved."
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