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A chemoenzymatic approach to the synthesis of enantiomerically pure (S)-3-hydroxy-γ-butyrolactone


Optically pure (S)-3-hydroxy-γ-butyrolactone, an important chiral building block in the pharmaceutical industry, was synthesized from l-malic acid by combining a selective hydrogenation and a lipase-catalyzed hydrolysis. Lipase from Candida rugosa was found to be the most efficient enzyme for the hydrolysis of (S)-β-benzoyloxy-γ-butyrolactone. The use of organic solvent-aqueous two-phase system was employed to extract benzoic acid generated from enzymatic hydrolysis of the substrate. Tert-butyl methyl ether as an organic solvent was effective to extract the reaction product, benzoic acid, and stably maintained the enzyme activity of Lipase OF immobilized on polymeric supports Amberlite® XAD-7. The immobilization made the recovery of the product simpler and prevented the formation of the emulsion. The pH adjustment was unnecessary with the immobilized Lipase OF. The scale-up of the enzymatic hydrolysis of S-BBL at 1,850-kg scale was carried out without problems to give 728.5kg of S-HGB at 80% isolated yield. The scale-up results are similar to those of bench scale reactions. Racemic (R,S)-β-benzoyloxy-γ-butyrolactone was prepared from d-, l-malic acid and was found to be hydrolyzed nonselectively by the enzyme.

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  1. Barbayianni E, Fotakopoulou I, Schmidt M, Constantinou-Kokotou V, Bornscheuer UT, Kokotos G (2005) Enzymatic removal of carboxyl protecting groups. 2. Cleavage of the benzyl and methyl moieties. J Org Chem 7:0:8730–8733

  2. Bevilaqua JV, Pinto JC, Limab LM, Barreiro EJ, Alves TLM, Freire DMG (2004) Enzymatic hydrolysis by immobilized lipase applied to a new prototype anti-asthma drug. Biochem Eng J 21:103–110

  3. Bornscheuer UT, Kazlauskas RJ (2006) Hydrolases in organic synthesis: regio- and stereoselective biotransformations, 2nd edn. Weinheim, Wiley-VCH

  4. Chikusa Y, Hirayama Y, Ikunaka M, Inoue T, Kamiyama S, Moriwaki M, Nishimoto Y, Nomoto F, Ogawa K, Ohno T, Otsuka K, Sakota AK, Shirasaka N, Uzura A, Uzura K (2003) There’s no industrial biocatalyst like hydrolase: development of scalable enantioselective processes using hydrolytic enzymes. Org Proc Res Dev 7:289–296

  5. Danda H, Maehara A, Umemura T (1991) Preparation of (4S)-4-hydroxy-3-methyl-2-(2′-propynyl)-2-cyclopentenone by combination of enzymatic hydrolysis and chemical transformation. Tetrahedron Lett 32:5119–5122

  6. Fernández-Lafuente R, Rodríguez V, Mateo C, Penzol G, Hernández-Justiz O, Irazoqui G, Villarino A, Ovsejevi K, Batista F, Guisán JM (1999) Stabilization of multimeric enzymes via immobilization and post-immobilization techniques. J Mol Catal B Enzym 7:181–189

  7. Fishman A, Eroshov M, Dee-Noor SS, van Mil J, Cogan U, Effenberger R (2001) A two step enzymatic process for large-scale production of (S)-and (R)-ethyl-3-hydroxybutyrate. Biotechnol Bioeng 74:256–263

  8. Fox ME, Jackson M, Lennon IC, McCague R, Parratt JS (2002) An enantioconvergent synthesis of (R)-4-acryloxy-1-butyne-3-ols for prosptanoid side chains. Adv Synth Catal 344:50–56

  9. Hollingworth RI, Wang G (2000) Toward a carbohydrate-based chemistry: progress in the development of general-purpose chiral synthons from carbohydrates. Chem Rev 100:4267–4282

  10. Hu S, Kelly S, Lee S, Tao J, Flahive E (2006) Efficient chemoenzymatic syntheis of pelitrexol via enzymatic differentiation of a remote stereocenter. Org Lett 8:1653–1655

  11. Jaeger K-E, Reetz MT (1998) Microbial lipases form versatile tools for biotechnology. Trends Biotechnol 16:396–403

  12. Kadereit D, Waldmann H (2001) Enzymatic protecting group techniques. Chem Rev 101:3367–3396

  13. Kataoka M, Hirakata M, Sakamoto K, Yamada H, Shimizu S (1996) Optical resolution of racemic pantoic acid through microbial stereoselective lactonization in an organic solvent/water two-phase system. Enzyme Microb Technol 19:307–310

  14. Kataoka M, Honda K, Shimizu S (2000) 3,4-Dihydrocoumarin hydrolase with haloperoxidase activity from Acinetobacter calcoaceticus F46. Eur J Biochem 267:3–10

  15. Khalameyzer V, Fischer I, Bornscheuer UT, Altenbuchner J (1999) Screening, nucleotide sequence, and biochemical characterization of an esterase from Pseudomonas fluorescens with high activity towards lactones. Appl Environ Microbiol 65:477–482

  16. Klibanov AM (1997) Why are enzymes less active in organic solvents than in water? Trends Biotechnol 15:97–101

  17. Kwak BS (2003) Development of chiral pharmaceutical fine chemicals through technology fusion. Chimica Oggi Jan–Feb:23–26

  18. Ladner W, Rettenmaier H, Zipperer B, Hansen H (1998) Process for the preparation of (L)-2-chloropropionic acid and its salts using lipase from Pseudomonas. US patent 5,753,495 (issued to BASF)

  19. Limanto J, Shafiee A, Devine PN, Upadhyay V, Desmond RA, Foster BR, Gauthier DR Jr, Reamer RA, Volante, RP (2005) An efficient chemoenzymatic approach to (S)- g-fluoroleucine ethyl ester. J Org Chem 70:2372–2375

  20. Muller M (2005) Chemoenzymatic synthesis of building blocks for statin side chains. Angew Chem Int Ed 44:362–365

  21. Nakagawa A, Idogaki H, Kato K, Shinmyo A, Suzuki T (2006) Improvement on production of (R)-4-chloro-3-hydroxybutyrate and (S)-3-hydroxy-g-butyrolactone with recombinant Escherichia coli cells. J Biosci Bioeng 101:97–103

  22. Palomo JM, Munoz G, Fernandez-Lorente G, Mateo C, Fernandez-Lafuente R, Guisan LM (2002) Interfacial adsorption of lipases on very hydrophobic support (octadecyl–Sepabeads): immobilization, hyperactivation and stabilization of the open form of lipases. J Mol Catal B Enzym 19–20:279–286

  23. Panke S, Wubbolts M (2002) The production of fine chemicals by biotransformations. Curr Opin Biotechnol 13:548–556

  24. Park OJ, Lee SH, Park TY, Chung WG, Lee SW (2006) Development of a scalable process for a key intermediate of (R)-Metalaxyl by enzymatic kinetic resolution. Org Proc Res Dev 10:588–591

  25. Sakamoto K, Honda K, Wada K, Kita S, Tsuzaki K, Nose H, Kataoka M, Shimizu S (2005) Practical resolution system for dl-pantoyl lactone using the lactonase from Fusarium oxysporum. J Biotechnol 118:99–106

  26. Schmidt M, Barbayianni E, Fotakopoulou I, Hohne M, Constantinou-Kokotou V, Bornscheuer UT, Kokotos G (2005) Enzymatic removal of carboxyl protecting groups. 1. Cleavage of the tert-butyl moiety. J Org Chem 7:0:3737–3740

  27. Segura RL, Palomo JM, Mateo C, Cortes A, Terreni M, Fernandez-Lafuente R, Guisan JM (2004) Different properties of the lipases contained in porcine pancreatic lipase extracts as enantioselective biocatalysts. Biotechnol Prog 20:825–829

  28. Sorgedrager MJ, Malpique R, van Rantwijk F, Sheldon RA (2004) Lipase catalysed resolution of nitro aldol adducts. Tetrahedron Asym 15:1295–1299

  29. Suzuki T, Idogaki H, Kasai N (1999) Dual production of highly pure methyl (R)-4-chloro-3-hydroxybutyrate and (S)-3-hydroxy-g-butyrolactone with Enterobacter sp. Enzyme Microb Technol 24:13–20

  30. Uh HS, Lee MJ (2002) Preparation of β-substituted γ-butyrolactone. KP 2002-0073751

  31. Veit T (2004) Biocatalysis for the production of cosmetic ingredients. Eng Life Sci 4:508–511

  32. Wang G, Hollingworth RI (2000) A simple three-step method for preparing homochiral 5-trityloxymethyl-2-oxazolidinones from optically active 3-hydroxy-γ-butyrolactones. Tetrahedron Asym 11:4429–4432

  33. Wells A (2006) What is in a biocatalyst? Org Proc Res Dev 10:678–681

  34. Yazbeck DR, Tao J, Martinez CA, Kline BJ, Hu S (2003) Automated enzyme screening methods for the preparation of enantiopure pharmaceutical intermediates. Adv Synth Catal 345:524–532

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We thank Miss. H.-J. Park for experimental help and Dr. K.-H. Cho and Dr. S.-W. Lee for their continued interest and support in this work.

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Correspondence to Oh-Jin Park.

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Lee, S., Park, O. & Uh, H. A chemoenzymatic approach to the synthesis of enantiomerically pure (S)-3-hydroxy-γ-butyrolactone. Appl Microbiol Biotechnol 79, 355–362 (2008). https://doi.org/10.1007/s00253-008-1439-9

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  • Lipases
  • Hydrolysis
  • (S)-3-hydroxy-γ-butyrolactone
  • Protection and deprotection
  • Two-phase
  • Candida rugosa