Tendulkar was bowled superbly by Peter Siddle for a dashing 73 from 98 deliveries, three balls before stumps at the MCG.
He had teamed up with Rahul Dravid, who scored a patient 68 not out to help the tourists reach 214 for three heading into the third day in response to Australia's first-innings score of 333.
The two greatest Test run-scorers of all-time put on 117 for the third wicket - their 20th century partnership.
Worryingly for Australia, they were looking at their ominous best before Siddle intervened and forced nightwatchman Ishant Sharma to make an appearance.
It did not take long for Tendulkar to get his eye in and he treated the 52,858-strong crowd to a batting masterclass with top-notch stroke play which delivered some perfectly-executed boundaries.
The 'Little Master' saw off eight balls in a nervous period before tea, following Virender Sehwag's dismissal for 67 in the penultimate over of the middle session.
He then set the tone for his innings with the first ball of the final session, doing so in spectacular style with an outrageous six when he flicked Siddle over the slip cordon and the third-man boundary.
It was the first of three precise late cuts he guided over the slips in what was a display of supreme awareness, vision and timing.
Tendulkar had eight fours to go with his six, and scored his 64th Test fifty with a single off Nathan Lyon that produced one of the loudest cheers of the day from a Melbourne crowd which featured a healthy portion of Indian supporters.
At the other end, Dravid was at his resolute best and lived up to his nickname of 'The Wall', with his 68 runs coming from 185 balls.
Dravid had been a lot more circumspect than Tendulkar but there were some flashes of brilliance, none more impressive than when he hit a glorious glance for four off Siddle after he opened his wrists ever so slightly.
Tendulkar overtook Dravid, who notched his 63rd Test fifty, despite the latter getting a 28-over head start.
Dravid was assured in his stroke-play and looked impregnable for the most part although he did experience some hairy moments courtesy of Ben Hilfenhaus, James Pattinson and Siddle.
Most notably, six overs before stumps, Siddle did find a way through when he bowled Dravid, but after much jubilation the delivery was adjudged to be a no-ball after umpire Marais Erasmus referred it to the third umpire, much to the Victorian firebrand's horror.
But Tendulkar's wicket would have served as a decent consolation for Siddle who bowled a ripping spell at the end of the day which yielded one for seven from four overs.
Tendulkar and Dravid had experienced little trouble dealing with the off-spin of Lyon, and the absence of an all-rounder in the team meant part-timers David Hussey and David Warner bowled their respective medium pace and leg spin with little effect against the greatest and second-greatest Test run scorers of all time.
Pacemen Pattinson and Hilfenhaus had made a blistering start to India's innings but could not build momentum.
Hilfenhaus bowled fantastically well early on and he troubled Dravid and Sehwag, in particular, on numerous occasions with superb pace, line and length.
While Pattinson was outstanding, especially in the period prior to tea which saw him take one for seven from five overs in a spell of venomous pace bowling which saw him bowl Sehwag after the opener had a number of near-misses in an entertaining and risky innings which included seven boundaries.
Sehwag brought up his 8,000th Test run and 31st Test fifty and combined well with Dravid as they put on 75 for the second wicket.
Earlier, Australia made it to 333 thanks to some handy cameos from Siddle (41), Hilfenhaus (19) and Pattinson (18 not out) after starting the day on 277 for six.
Veteran Indian paceman Zaheer Khan (four for 77) and off-spinner Ravichandran Ashwin (three for 81) wrapped up Australia's innings as they shared the remaining four wickets evenly.
Meanwhile, impressive Australian debutant Ed Cowan is under a fitness cloud after injuring his back during the warm-up and receiving treatment.
Cowan re-entered the arena to resume fielding duties in the second session but had to leave the field again soon after.
PA
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