Abstract
A reversible salicylic acid decarboxylase (Sdc) catalyzes the carboxylation of m-aminophenol (m-AP) to paminosalicylic acid (PAS) as an antituberculous agent, through an enzymatic KolbeSchmitt reaction. To develop a highyield PAS production system through such an enzymatic reaction, we generated Sdc mutants by site-directed mutagenesis and succeeded in generating several mutants showing increased carboxylation specific activities. Among them, a Y64T-F195Y-Sdc mutant showed a 12-fold higher carboxylation specific activity toward m-AP than wild-type Sdc. By the whole-cell reaction of recombinant Escherichia coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc, 70mM PAS was produced from 100 mM m-AP within 2 h. This reaction time was shortened to one-twelfth that of the PAS production using E. coli BL21(DE3) expressing the gene encoding wild-type Sdc (24 h). Moreover, 140 mM PAS was produced from 200 mM m-AP within 9 h by the whole-cell reaction of recombinant E. coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc.
| Original language | English |
|---|---|
| Pages (from-to) | 628-634 |
| Number of pages | 7 |
| Journal | Bulletin of the Chemical Society of Japan |
| Volume | 86 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2013 |
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P-aminosalicylic acid production by enzymatic kolbeschmitt reaction using salicylic acid decarboxylases improved through site-directed mutagenesis. / Ienaga, Saori; Kosaka, Sachiyo; Honda, Yuki; Ishii, Yoshitaka; Kirimura, Kotaro.
In: Bulletin of the Chemical Society of Japan, Vol. 86, No. 5, 2013, p. 628-634.Research output: Contribution to journal › Article
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TY - JOUR
T1 - P-aminosalicylic acid production by enzymatic kolbeschmitt reaction using salicylic acid decarboxylases improved through site-directed mutagenesis
AU - Ienaga, Saori
AU - Kosaka, Sachiyo
AU - Honda, Yuki
AU - Ishii, Yoshitaka
AU - Kirimura, Kotaro
PY - 2013
Y1 - 2013
N2 - A reversible salicylic acid decarboxylase (Sdc) catalyzes the carboxylation of m-aminophenol (m-AP) to paminosalicylic acid (PAS) as an antituberculous agent, through an enzymatic KolbeSchmitt reaction. To develop a highyield PAS production system through such an enzymatic reaction, we generated Sdc mutants by site-directed mutagenesis and succeeded in generating several mutants showing increased carboxylation specific activities. Among them, a Y64T-F195Y-Sdc mutant showed a 12-fold higher carboxylation specific activity toward m-AP than wild-type Sdc. By the whole-cell reaction of recombinant Escherichia coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc, 70mM PAS was produced from 100 mM m-AP within 2 h. This reaction time was shortened to one-twelfth that of the PAS production using E. coli BL21(DE3) expressing the gene encoding wild-type Sdc (24 h). Moreover, 140 mM PAS was produced from 200 mM m-AP within 9 h by the whole-cell reaction of recombinant E. coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc.
AB - A reversible salicylic acid decarboxylase (Sdc) catalyzes the carboxylation of m-aminophenol (m-AP) to paminosalicylic acid (PAS) as an antituberculous agent, through an enzymatic KolbeSchmitt reaction. To develop a highyield PAS production system through such an enzymatic reaction, we generated Sdc mutants by site-directed mutagenesis and succeeded in generating several mutants showing increased carboxylation specific activities. Among them, a Y64T-F195Y-Sdc mutant showed a 12-fold higher carboxylation specific activity toward m-AP than wild-type Sdc. By the whole-cell reaction of recombinant Escherichia coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc, 70mM PAS was produced from 100 mM m-AP within 2 h. This reaction time was shortened to one-twelfth that of the PAS production using E. coli BL21(DE3) expressing the gene encoding wild-type Sdc (24 h). Moreover, 140 mM PAS was produced from 200 mM m-AP within 9 h by the whole-cell reaction of recombinant E. coli BL21(DE3) expressing the gene encoding Y64T-F195Y-Sdc.
UR - http://www.scopus.com/inward/record.url?scp=84878886695&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84878886695&partnerID=8YFLogxK
U2 - 10.1246/bcsj.20130006
DO - 10.1246/bcsj.20130006
M3 - Article
AN - SCOPUS:84878886695
VL - 86
SP - 628
EP - 634
JO - Bulletin of the Chemical Society of Japan
JF - Bulletin of the Chemical Society of Japan
SN - 0009-2673
IS - 5
ER -