Tripura Transport Minister Manik Dey flagged off the bus service at the Tripura Road Transport Corp (TRTC) international bus terminus depot.
A 45-seater Volvo bus, carrying 18 passengers including two Canadians, left in the afternoon for Kolkata via Dhaka. Six of the passengers were bound for Kolkata and 12 for Bangladesh.
Prime Minister Narendra Modi and his Bangladeshi counterpart Sheikh Hasina jointly flagged off the Dhaka-Shillong-Guwahati and Kolkata-Dhaka-Agartala bus services during Modi's visit to Bangladesh on June 6.
While the Dhaka-Shillong-Guwahati service is yet to start, the Kolkata-Agartala via Dhaka service began operating but with only the West Bengal and Bangladesh operating their buses.
On Friday, Tripura's buses joined the service.
The buses, scheduled to ply twice a week by each transporter, have a fare ranging between Rs.1,800 and Rs.2,000.
The distance between Kolkata and Agartala is about 1,650 km if one skirts Bangladesh. But the distance falls to 515 km if the buses go through Dhaka.
Due to non-availability of air tickets and for other reasons, the buses have become a popular mode of transport between West Bengal and Tripura via the Bangladesh capital.
The Kolkata-Dhaka bus link began in June 1999. And a separate Agartala-Dhaka service started in September 2003.
After the roughing up of a Tripura official in Dhaka on Monday, the Indian government has asked Bangladesh to provide adequate security to the bus service, TRTC managing director Keshab Kar said.
He said TRTC liaison officer L.J. Rokhum was roughed up by unidentified people when he went to Dhaka to get road permit for the new Volvo bus.
Occasionally, political turmoil in Bangladesh has hit the bus services between the two countries.