Ross Whiteley made more useful runs and picked up a rare wicket but his side Sylhet Sixers suffered their first defeat in the Bangladesh Premier League.
The Worcestershire all-rounder was second top scorer with 27 and returned 1-10 from his two overs.
It followed on from his 25 not out off just 12 balls – with three sixes – in the win over Rajshahi Kings.
But the Sixers, who had won their opening three games, went down by six wickets as Khulna overhauled their total of 135-5 on a sluggish pitch with two overs to spare.
Sylhet were put into bat and Whiteley came to the wicket at 67-4 in the seventh over and added 57 in seven overs with Nasir Hossain (47 not out).
Whiteley struck Sussex paceman Jofra Archer for his only six over long off.
But, after making his runs off 23 deliveries, he was dismissed by the same bowler when caught off a miscue at deep square leg.
Three quick wickets from Taijul Islam gave the Sixers hopes of retaining their 100 per cent record as the Titans were reduced to 43-3.
Whiteley then enjoyed his moment of success with the ball as Titans skipper Mahmudullah (27) holed out at long on at 93-4.
But Gloucestershire’s Michael Klinger, with an unbeaten 47, and Carlos Brathwaite (23 not out) saw Khulna to 138-4 in 18 overs.