Improvements of enzyme activity and enantioselectivity in lipase-catalyzed alcoholysis of (R,S)-azolides

Abstract With Candida antarctica lipase B (CALB)-catalyzed alcoholysis of (R,S)-naproxenyl 1,2,4-triazolide at the optimal conditions (i.e. anhydrous MTBE as the solvent, and methanol as the acyl acceptor at 45 °C) as the model system, the enzyme enantioselectivity in terms of VR/VS = 105.8 and specific activity for the fast-reacting (R)-azolide VR/(Et) = 0.979 mmol/(h g) were greatly improved in comparison with VR/VS = 8.0 and VR/(Et) = 0.113 mmol/(h g) of using (R,S)-naproxenyl 2,2,2-trifluoroethyl ester as the substrate. The resolution strategy was successfully extended to other (R,S)-profenyl 1,2,4-triazolides and lipases from Candida rugosa (Lipase MY) and Carica papaya (CPL) having opposite enantioselectivity to CALB. Moreover, the kinetic constants were estimated, compared with those obtained via hydrolysis, and employed for modeling time-course conversions of (R,S)-naproxenyl 1,2,4-triazolide in anhydrous MTBE. The advantages of easy substrate preparation, high enzyme reactivity and enantioselectivity, as well as easy product separation from the remaining substrate via reactive extraction demonstrate merits of using (R,S)-azolides but not the corresponding esters for the alcoholytic resolution.

[1]  U. Bornscheuer,et al.  Hydrolases in Organic Synthesis: Regio- and Stereoselective Biotransformations , 1999 .

[2]  S. Tsai,et al.  Improvements of enzyme activity and enantioselectivity via combined substrate engineering and covalent immobilization , 2008, Biotechnology and bioengineering.

[3]  V. Gotor‐Fernández,et al.  Enzymatic Aminolysis and Ammonolysis Processes in the Preparation of Chiral Nitrogenated Compounds , 2006 .

[4]  I. Alfonso,et al.  Asymmetric organic synthesis with enzymes , 2008 .

[5]  K. Hult,et al.  Understanding water effect on Candida antarctica lipase B activity and enantioselectivity towards secondary alcohols , 2009 .

[6]  E. P. Hudson,et al.  Biocatalysis in semi-aqueous and nearly anhydrous conditions. , 2005, Current opinion in biotechnology.

[7]  R. Chěnevert,et al.  Stereoselective Enzymatic Acylations (Transesterifications) , 2006 .

[8]  Bernhard Hauer,et al.  Industrial methods for the production of optically active intermediates. , 2004, Angewandte Chemie.

[9]  U. Hanefeld.  Reagents for (ir)reversible enzymatic acylations. , 2003, Organic & biomolecular chemistry.

[10]  S. Tsai,et al.  Effects of hydrolysis and esterification side-reactions on the kinetic resolution of enzyme-catalyzed irreversible transesterification in organic solvents , 2000 .

[11]  I. Ng,et al.  Implication of substrate-assisted catalysis on improving lipase activity or enantioselectivity in organic solvents. , 2006, Biochimica et biophysica acta.

[12]  B. G. Davis.  Chemical modification of biocatalysts. , 2003, Current opinion in biotechnology.

[13]  H. Staab,et al.  Azolides In Organic Synthesis And Biochemistry , 1998 .

[14]  A. Patti,et al.  Resolution of racemic flurbiprofen by lipase-mediated esterification in organic solvent , 1995 .

[15]  F. Theil.  Enhancement of Selectivity and Reactivity of Lipases by Additives , 2000 .

[16]  U. Bornscheuer,et al.  Lipase-Catalyzed Resolution of Ibuprofen , 2000 .

[17]  S. Tsai,et al.  Resolution of non-protein amino acids via Carica papaya lipase-catalyzed enantioselective transester , 2005 .

[18]  Sajja Hari Krishna,et al.  Optimizing lipases and related enzymes for efficient application. , 2002, Trends in biotechnology.

[19]  M. Reetz.  Directed Evolution of Enantioselective Enzymes as Catalysts for Organic Synthesis , 2006 .

[20]  E. Santaniello,et al.  Lipase-Catalyzed Deacylation by Alcoholysis: A Selective, Useful Transesterification Reaction , 2006 .

[21]  M. Fahim,et al.  Liquid–liquid equilibria of water + MTBE + reformate , 2001 .

[22]  Donald Hilvert,et al.  Minimalist active-site redesign: teaching old enzymes new tricks. , 2007, Angewandte Chemie.

[23]  S. Tsai,et al.  (R,S)‐Azolides as Novel Substrates for Lipase‐Catalyzed Hydrolytic Resolution in Organic Solvents , 2009 .

[24]  S. Tsai,et al.  Enzymatic resolution of (RS)-2-arylpropionic acid thioesters by Candida rugosa lipase-catalyzed thiotransesterification or hydrolysis in organic solvents , 1998 .