Cytochrome P450 (CYP or P450) refers to a various protein that catalyze the oxidation of organic substances. Plant CYPs catalyze a variety of biochemical reactions, including the synthesis of various hormones, defense substances and secondary metabolites associated with biological and environmental stresses. However, only a limited number of CYPs in plants have been investigated at the molecular level. To expand our understanding of the biological functions of SlP19 family, consisting of SlP19-1, 2, and 3 was isolated from the tomato genome. SlP19-1, 2, and 3, showed distinct expression patterns in various tomato tissues. Subcellular localization analysis showed that SlP19-1, 2 and 3, similar to other known CYPs, were found in the endoplasmic reticulum. SlP19-1, 2, and 3 were heterologously expressed in Escherichia coli to determine enzymatic properties of the selected SlP19s, Isolated SlP19-1, 2, and 3 proteins catalyzed 7-ethoxycoumarin hydroxylation. Additional NADPH oxidation assay revealed that SlP19-1, 2, and 3 showed a catalytic activity toward volatile terpenes, including trans-nerolidol and geranyllinalool, which are known to be cleaved to form an active component upon wounding. The expression levels of SlP19-1 and SlP19-2 increased when wound stress was applied to tomato. These results suggested that SlP19 mediates the production of volatile signaling molecules within the wound in response to mechanical attacks, which activates wound respons in tomato plants, and SlP19 is presumed to be responsible for producing volatile substance that induce wounding signal using sesquiterpenes and monoterpenes as substrates.