The veteran actor has knocked Priyanka Chopra into second place with over 1,850,903 followers on the microblogging site, Economic Times reports.
Chopra was the most popular Hindi film star on Twitter, but in recent months the star has tweeted less as she completes three films and records her debut album.
Bachchan has shared over 11,108 posts since he began tweeting in 2010.
The Silsila star also maintains a blog on Big Adda.
Bachchan also uses voice blogging to keep in touch with followers through his Bachchan Bol subscribed mobile phone service available in India and in the UK.
The actor recently criticised Chopra for retweeting an F-word tweet originally written by Rihanna.