Roberto Rosales' emphatic header brought Twente level again only for Spurs to forge back in front as a defensive slip allowed Wilson Palacios in on goal and when his shot was saved Defoe was on hand to fire home the loose ball.
However, the goals continued to flow with Nacer Chadli leaving Heurelho Gomes grasping at thin air with a delightful free-kick in the 64th minute as Twente headed for the Europa League.
Manchester United finally saw their defence breached but hit back against Valencia to win Group C courtesy of a 1-1 draw at Old Trafford.
United's hopes of becoming the first club ever to go through a Champions League group phase without conceding a goal were ended when Michael Carrick gave the ball away to Alejandro Dominguez who found Pablo Hernandez to fire a low shot beyond Ben Amos in the 32nd minute.
United were back on terms midway through the second period when Park Ji-Sung's powerful strike was only parried by Vicente Guaita and Anderson was on hand to volley home.
Rangers and Bursaspor knew long before their 1-1 draw in Turkey that they were certain to finish third and fourth, respectively.
Gers took the lead through a familiar source in the 19th minute when Kenny Miller smashed the ball home in style after being found in the box by Steven Whittaker's low cross from the left.
But their hopes of victory were dashed 11 minutes from time when Sercan Yildirim was found six yards from goal and had time to slot home.
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