Advertisement

Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Regioselective carboxylation of 1,3-dihydroxybenzene by 2,6-dihydroxybenzoate decarboxylase of Pandoraea sp. 12B-2

Abstract

We found a bacterium, Pandoraea sp. 12B-2, of which whole cells catalyzed not only the decarboxylation of 2,6-dihydroxybenzoate but also the regioselective carboxylation of 1,3-dihydroxybenzene to 2,6-dihydroxybenzoate. The whole cells of Pandoraea sp. 12B-2 also catalyzed the regioselective carboxylation of phenol and 1,2-dihydroxybenzene to 4-hydroxybenzoate and 2,3-dihydroxybenzoate, respectively. The molar conversion ratio of the carboxylation reaction depended on the concentration of KHCO3 in the reaction mixture. Only 5 or 48 % of 1,3-dihydroxybenzene added was converted into 2,6-dihydroxybenzoate in the presence of 0.1 M or 3 M KHCO3, respectively. The addition of acetone to the reaction mixture increased the initial rate of the carboxylation reaction, but the final molar conversion yield reached almost the same value. When the efficient production of 2,6-dihydroxybenzoate was optimized using the whole cells of Pandoraea sp. 12B-2, the productivity of 2,6-dihydroxybenzoate topped out at 1.43 M, which was the highest value so far reported. No formation of any other products was observed after the carboxylation reaction.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Scheme 1

References

  1. Anderson JJ, Dagley S (1981) Catabolism of tryptophan, anthranilate, and 2,3-dihydroxybenzoate in Trichosporon cutaneum. J Bacteriol 146:291–297

  2. Brackmann R, Fuchs G (1993) Enzymes of anaerobic metabolism of phenolic compounds 4-hydroxybenzoil-CoA reductase (dehydroxylating) from a denitrifying Pseudomonas species. Eur J Biochem 213:563–571

  3. He Z, Wiegel J (1995) Purification and characterization of an oxygen-sensitive reversible 4-hydroxybenzoate decarboxylase from Clostridium hydroxybenzoicum. Eur J Biochem 229:77–82

  4. He Z, Wiegel J (1996) Purification and characterization of an oxygen-sensitive 3,4-dihydroxybenzoate decarboxylase from Clostridium hydroxybenzoicum. J Bacteriol 178:3539–3543

  5. Ishii Y, Narimatsu Y, Iwasaki Y, Arai N, Kino K, Kirimura K (2004) Reversible and nonoxidative γ-resorcinolic acid decarboxylase: characterization and gene cloning of a novel enzyme catalyzing carboxylation of resorcinol, 1,3-dihydroxybenzene, from Rhizobium radiobacter. Biochem Biophys Res Commun 324:611–620

  6. Kamath AV, Dasgupta D, Vaidyanathan CS (1987) Enzyme-catalysed non-oxidative decarboxylation of aromatic acids. I. Purification and spectroscopic properties of 2,3-dihydroxybenzoic acid decarboxylase from Aspergillus niger. Biochem Biophys Res Commun 145:586–595

  7. Matsuda T, Ohashi Y, Harada T, Yanagihara Y, Nagasawa T, Nakamura K (2001) Conversion of pyrrole to pyrrole-2-carboxylate by cells of Bacillus megaterium in supercritical CO2. Chem Commun (Camb) 21:2194–2195

  8. Nakamatsu T, Nishida Y, Kometani N (1993) Method for producing 2,6-dihydroxybenzoic acid. Patent EP 0552912 A2

  9. Omura H, Wiser M, Nagasawa T (1998) Pyrrole-2-carboxylate decarboxylase from Bacillus megaterium PYR2910, an organic-acid-requiring enzyme. Eur J Biochem 253:480–484

  10. Sagawa M, Okai M, Mizutani B, Takaba H, Ue M (2000) Display device. Patent JP 12323016 A

  11. Santha R, Rao NA, Vaidyanathan CS (1996) Identification of the active-site peptide of 2,3-dihydroxybenzoic acid decarboxylase from Aspergillus oryzae. Biochim Biophys Acta 1293:191–200

  12. Toshimoto M (1996) Heat-developable diazo copying material. Patent JP 08272035 A

  13. Uchida A, Ogawa M, Yoshida T, Nagasawa T (2003) Quinolinate dehydrogenase and 6-hydroxyquinolinate decarboxylase involved in the conversion of quinolinic acid to 6-hydroxynicotinic acid by Alcaligenes sp. UK21. Arch Microbiol 180:81–87

  14. Wieser M, Fujii N, Yoshida T, Nagasawa T (1998) Carbon dioxide fixation by reversible pyrrole-2-carboxylate decarboxylase from Bacillus megaterium PYR2910. Eur J Biochem 257:495–499

  15. Yoshida T, Fujita K, Nagasawa T (2002) Novel reversible indole-3-carboxylate decarboxylase catalyzing nonoxidative decarboxylation. Biosci Biotechnol Biochem 66:2388–2394

  16. Yoshida T, Hayakawa Y, Matsui T, Nagasawa T (2004a) Purification and characterization of 2,6-dihydroxybenzoate decarboxylase reversibly catalyzing nonoxidative decarboxylation. Arch Microbiol 181:391–397

  17. Yoshida M, Fukuhara N, Oikawa T (2004b) Thermophilic, reversible γ-resorcylate decarboxylase from Rhizobium sp. strain MTP-10005: purification, molecular characterization, and expression. J Bacteriol 186:6855–6863

  18. Zhang X, Wiegel J (1994) Reversible conversion of 4-hydroxybenzoate and phenol by Clostridium hydroxybenzoicum. Appl Environ Microbiol 60:4182–4185

Download references

Acknowledgements

This study was carried out as part of The Project for Development of a Technological Infrastructure for Industrial Bioprocesses on R&D of New Industrial Science and Technology Frontiers by Ministry of Economy, Trade & Industry (METI), and entrusted by the New Energy and Industrial Technology Development Organization (NEDO).

Author information

Correspondence to Toru Nagasawa.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Matsui, T., Yoshida, T., Yoshimura, T. et al. Regioselective carboxylation of 1,3-dihydroxybenzene by 2,6-dihydroxybenzoate decarboxylase of Pandoraea sp. 12B-2. Appl Microbiol Biotechnol 73, 95–102 (2006). https://doi.org/10.1007/s00253-006-0437-z

Download citation

Keywords

  • KHCO3
  • Decarboxylase Activity
  • Diisopropyl Ether
  • Standard Reaction Mixture
  • Molar Conversion