Thisbook constitutes the refereed proceedings of the 16th European Conference onEvolutionary Computation in Combinatorial Optimization, EvoCOP 2016, held in Porto,Portugal, in March/April 2016, co-located with the Evo*2015 events EuroGP,EvoMUSART and EvoApplications.The17 revised full papers presented were carefully reviewed and selected from 44submissions. The papers cover methodology, applications and theoretical studies. Themethods included evolutionary and memetic algorithms, variable neighborhoodsearch, particle swarm optimization, hyperheuristics, mat-heuristic and otheradaptive approaches. Applications included both traditional domains, such asgraph coloring, vehicle routing, the longest common subsequence problem, thequadratic assignment problem; and new(er) domains such as the traveling thiefproblem, web service location, and finding short addition chains. Thetheoretical studies involved fitness landscape analysis, local search and recombinationoperator analysis, and the big valley search space hypothesis. Theconsideration of multiple objectives, dynamic and noisy environments was alsopresent in a number of articles.