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In enzymology, a 3-hydroxypalmitoyl-[acyl-carrier-protein] dehydratase (EC 4.2.1.61) is an enzyme that catalyzes the chemical reaction

(3R)-3-hydroxypalmitoyl-[acyl-carrier-protein] ${\displaystyle \rightleftharpoons }$ hexadec-2-enoyl-[acyl-carrier-protein] + H₂O

Hence, this enzyme has one substrate, [[(3R)-3-hydroxypalmitoyl-[acyl-carrier-protein]]], and two products, [[hexadec-2-enoyl-[acyl-carrier-protein]]] and H₂O.

This enzyme belongs to the family of lyases, specifically the hydro-lyases, which cleave carbon-oxygen bonds. The systematic name of this enzyme class is (3R)-3-hydroxypalmitoyl-[acyl-carrier-protein] hydro-lyase (hexadec-2-enoyl-[acyl-carrier protein]-forming). Other names in common use include D-3-hydroxypalmitoyl-[acyl-carrier-protein] dehydratase, beta-hydroxypalmitoyl-acyl carrier protein dehydrase, beta-hydroxypalmitoyl thioester dehydratase, beta-hydroxypalmityl-ACP dehydrase, and (3R)-3-hydroxypalmitoyl-[acyl-carrier-protein] hydro-lyase. This enzyme participates in fatty acid biosynthesis.

As of late 2007, only one structure has been solved for this class of enzymes, with the PDB accession code 2PFF.

• Mizugaki M, Swindell AC, Wakil SJ (1968). "Intermediate- and long-chain beta-hydroxyacyl-ACP dehydrases from E coli fatty acid synthetase". Biochem. Biophys. Res. Commun. 33 (3): 520–7. doi:10.1016/0006-291X(68)90607-4. PMID 4881058.