TY - JOUR
T1 - Expanding substrate specificity of salicylate decarboxylase by site-directed mutagenesis for expansion of the entrance region connecting to the substrate access tunnel
AU - Kirimura, Kotaro
AU - Araki, Masahiro
AU - Ishihara, Mana
AU - Ishii, Yoshitaka
PY - 2019/1/1
Y1 - 2019/1/1
N2 - Salicylate decarboxylase (Sdc, EC 4.1.1.91) from the yeast Trichosporon moniliiforme WU-0401 catalyzes the carboxylation of phenol to salicylic acid and is applicable to enzymatic Kolbe-Schmitt reaction. Based on the 3D-modeling of Sdc, we generated a Sdc mutant, K23A-Sdc, by site-directed mutagenesis for expansion of the entrance region connecting to the substrate access tunnel. K23A-Sdc catalyzed the carboxylation of o-cresol to 3-methyl salicylic acid (3-MS), whereas Sdc showed negligible and slight activity toward o-cresol and 3-MS, respectively. Simultaneously, K23A-Sdc showed approximately 5.8-fold more decarboxylation activity toward 3-MS than Sdc. These results clearly indicate that K23A-Sdc acquired novel substrate specificity by substitution of only one amino acid residue (23rd lysine residue to alanine residue), around the substrate entrance region but not at the active center of Sdc.
AB - Salicylate decarboxylase (Sdc, EC 4.1.1.91) from the yeast Trichosporon moniliiforme WU-0401 catalyzes the carboxylation of phenol to salicylic acid and is applicable to enzymatic Kolbe-Schmitt reaction. Based on the 3D-modeling of Sdc, we generated a Sdc mutant, K23A-Sdc, by site-directed mutagenesis for expansion of the entrance region connecting to the substrate access tunnel. K23A-Sdc catalyzed the carboxylation of o-cresol to 3-methyl salicylic acid (3-MS), whereas Sdc showed negligible and slight activity toward o-cresol and 3-MS, respectively. Simultaneously, K23A-Sdc showed approximately 5.8-fold more decarboxylation activity toward 3-MS than Sdc. These results clearly indicate that K23A-Sdc acquired novel substrate specificity by substitution of only one amino acid residue (23rd lysine residue to alanine residue), around the substrate entrance region but not at the active center of Sdc.
KW - Enzymatic Kolbe-Schmitt reaction
KW - Salicylate decarboxylase
KW - Substrate specificity
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U2 - 10.1246/cl.180755
DO - 10.1246/cl.180755
M3 - Article
AN - SCOPUS:85059425501
VL - 48
SP - 58
EP - 61
JO - Chemistry Letters
JF - Chemistry Letters
SN - 0366-7022
IS - 1
ER -