In enzymology, a thiophene-2-carbonyl-CoA monooxygenase (EC 1.14.99.35) is an enzyme that catalyzes the chemical reaction

thiophene-2-carbonyl-CoA + AH₂ + O₂ ${\displaystyle \rightleftharpoons }$ 5-hydroxythiophene-2-carbonyl-CoA + A + H₂O

The three substrates of this enzyme are thiophene-2-carbonyl-CoA, an electron acceptor AH₂, and O₂. Its three products are 5-hydroxythiophene-2-carbonyl-CoA, the reduction product A, and H₂O.

This enzyme belongs to the family of oxidoreductases, specifically those acting on paired donors, with O₂ as oxidant and incorporation or reduction of oxygen. The oxygen incorporated need not be derived from O miscellaneous. The systematic name of this enzyme class is thiophene-2-carbonyl-CoA, hydrogen-donor:oxygen oxidoreductase. Other names in common use include thiophene-2-carboxyl-CoA dehydrogenase, thiophene-2-carboxyl-CoA hydroxylase, and thiophene-2-carboxyl-CoA monooxygenase.