Modulation of Immobilized Lipase Enantioselectivity via Chemical Amination

The aspartic and glutamic carboxylic groups of the surface of three different immobilized lipases (those from Candida antarctica (form B) (CAL-B), Thermomyces lanuginose (TLL) and Pseudomonas fluorescens (PFL) have been modified with ethylenediamine (after activation of the carboxylic function with carbodiimide). The exchange of groups with a negative charge for positively charged groups permitted us to greatly alter the activity, specificity and stereoselectivity of the lipase. Thus, in some cases, the activity of the lipases increased after the chemical modification while in other cases the activity was strongly reduced. Moreover, the effect of the experimental conditions on the activity was also strongly altered. Remarkably, the enantioselectivity of the enzymes in the hydrolysis of different mandelic acid derivatives was strongly modulated. For example, amination of the CNBr-CAL-B preparation greatly increased the enantioselectivity of the enzyme in the hydrolysis of (±)-2-hydroxyphenylacetic acid methyl ester, from an E value of 2 without modification up to E>100, affording (R)-mandelic acid in high purity (ee>99 % at 50 % conversion) at pH 7 and 4 °C. Thus, the chemical modification of lipases seems to be a very powerful tool to improve their performance as enantioselective biocatalysts.

[1]  K. Dettner,et al.  Chemical and enzymatic synthesis of buprestin A and B—bitter acylglucosides from Australian jewel beetles (Coleoptera: Buprestidae) , 2006 .

[2]  R. Fernández-Lafuente,et al.  Chemical modification of protein surfaces to improve their reversible enzyme immobilization on ionic exchangers. , 2006, Biomacromolecules.

[3]  J. Cooney,et al.  Direct acylation of flavonoid glycosides with phenolic acids catalysed by Candida antarctica lipase B (Novozym 435 , 2006 .

[4]  A. Petit,et al.  Preparation of various enantiomerically pure (benzotriazol-1-yl)- and (benzotriazol-2-yl)-alkan-2-ols , 2006 .

[5]  A. Chaubey,et al.  Enantioselectivity modulation through immobilization of Arthrobacter sp. lipase: Kinetic resolution of fluoxetine intermediate , 2006 .

[6]  V. Wray,et al.  Production and modification of bioactive biosurfactants , 2006 .

[7]  C. Ortiz,et al.  Glutaraldehyde cross-linking of lipases adsorbed on aminated supports in the presence of detergents leads to improved performance. , 2006, Biomacromolecules.

[8]  R. Fernández-Lafuente,et al.  Purification and identification of different lipases contained in PPL commercial extracts: A minor contaminant is the main responsible of most esterasic activity , 2006 .

[9]  R. Fernández-Lafuente,et al.  Effect of lipase–lipase interactions in the activity, stability and specificity of a lipase from Alcaligenes sp. , 2006 .

[10]  Y. Kita,et al.  A dynamic kinetic resolution of allyl alcohols by the combined use of lipases and [VO(OSiPh3)3]. , 2006, Angewandte Chemie.

[11]  S. M. Glueck,et al.  Enantio-complementary deracemization of (±)-2-hydroxy-4-phenylbutanoic acid and (±)-3-phenyllactic acid using lipase-catalyzed kinetic resolution combined with biocatalytic racemization , 2006 .

[12]  M. Ueda,et al.  Enhancement of Activity of Lipase‐Displaying Yeast Cells and Their Application to Optical Resolution of (R,S)‐1‐Benzyloxy‐3‐Chloro‐2‐Propyl Monosuccinate , 2006, Biotechnology progress.

[13]  C. Ortiz,et al.  Purification of different lipases from Aspergillus niger by using a highly selective adsorption on hydrophobic supports. , 2005, Biotechnology and bioengineering.

[14]  U. Bornscheuer,et al.  Optimization of lipase-catalyzed glucose fatty acid ester synthesis in a two-phase system containing ionic liquids and t-BuOH , 2005 .

[15]  R. Volante,et al.  An efficient enzyme-catalyzed kinetic resolution: large-scale preparation of an enantiomerically pure indole-ethyl ester derivative, a key component for the synthesis of a prostaglandin D2 receptor antagonist, an anti-allergic rhinitis drug candidate , 2005 .

[16]  O. Abián,et al.  Some special features of glyoxyl supports to immobilize proteins , 2005 .

[17]  J. Karlsson,et al.  Enzymatic kinetic resolution of 1-(3-furyl)-3-buten-1-ol , 2005 .

[18]  C. Ortiz,et al.  Lipase–lipase interactions as a new tool to immobilize and modulate the lipase properties , 2005 .

[19]  D. Reyes-Duarte,et al.  Synthesis of sugar esters in solvent mixtures by lipases from Thermomyces lanuginosus and Candida antarctica B, and their antimicrobial properties , 2005 .

[20]  R. Fernández-Lafuente,et al.  Synthesis of enantiomerically pure glycidol via a fully enantioselective lipase-catalyzed resolution , 2005 .

[21]  A. Ghanem,et al.  One-pot synthesis and chiral analysis of cyclopropane derivatives. , 2005, Chirality.

[22]  R. Fernández-Lafuente,et al.  Purification, Immobilization, and Stabilization of a Lipase from Bacillus thermocatenulatus by Interfacial Adsorption on Hydrophobic Supports , 2008, Biotechnology progress.

[23]  S. Lutz.  Engineering lipase B from Candida antarctica , 2004 .

[24]  N. Krieger,et al.  Hydrolysis and synthesis reactions catalysed by Thermomyces lanuginosa lipase in the AOT/Isooctane reversed micellar system , 2004 .

[25]  C. Ortiz,et al.  Use of immobilized lipases for lipase purification via specific lipase-lipase interactions. , 2004, Journal of chromatography. A.

[26]  C. Ching,et al.  Enhanced activity and enantioselectivity of Candida rugosa lipase immobilized on macroporous adsorptive resins for ibuprofen resolution , 2004, Biotechnology Letters.

[27]  R. Fernández-Lafuente,et al.  Resolution of paroxetine precursor using different lipases: Influence of the reaction conditions on the enantioselectivity of lipases , 2004 .

[28]  C. Keith,et al.  Highly enantioselective kinetic resolution of primary alcohols of the type Ph-X-CH(CH3)-CH2OH by Pseudomonas cepacia lipase: effect of acyl chain length and solvent , 2003 .

[29]  R. Fernández-Lafuente,et al.  Self-assembly of Pseudomonas fluorescens lipase into bimolecular aggregates dramatically affects functional properties. , 2003, Biotechnology and bioengineering.

[30]  R. Fernández-Lafuente,et al.  General trend of lipase to self-assemble giving bimolecular aggregates greatly modifies the enzyme functionality. , 2003, Biomacromolecules.

[31]  C. Ortiz,et al.  Modulation of the enantioselectivity of lipases via controlled immobilization and medium engineering: hydrolytic resolution of mandelic acid esters , 2002 .

[32]  J. Bäckvall,et al.  Efficient Lipase‐Catalyzed Kinetic Resolution and Dynamic Kinetic Resolution of β‐Hydroxy Nitriles. Correction of Absolute Configuration and Transformation to Chiral β‐Hydroxy Acids and γ‐Amino Alcohols , 2002 .

[33]  R. Fernández-Lafuente,et al.  Modulation of the enantioselectivity of Candida antarctica B lipase via conformational engineering. Kinetic resolution of (±)-α-hydroxy-phenylacetic acid derivatives , 2002 .

[34]  Satwinder Singh,et al.  Kinetic resolution of heteroaryl β-hydroxy sulfides catalyzed by Humicola lanuginosa lipase , 2002 .

[35]  S. Riva.  Biocatalytic modification of natural products. , 2001, Current opinion in chemical biology.

[36]  D. Lawson,et al.  Structural origins of the interfacial activation in Thermomyces (Humicola) lanuginosa lipase. , 2000, Biochemistry.

[37]  J. Vatèle,et al.  TOTAL SYNTHESIS OF ANTITUMOR GONIOTHALAMUS STYRYLLACTONES , 1999 .

[38]  Y. Khmelnitsky,et al.  Biocatalytic combinatorial synthesis. , 1999, Bioorganic & medicinal chemistry.

[39]  D. Clark,et al.  Combinatorial biocatalysis: taking the lead from nature. , 1999, Current opinion in biotechnology.

[40]  J. Vatèle,et al.  First total synthesis of (−)-8-epi-9-deoxygoniopypyrone , 1998 .

[41]  D. Clark,et al.  Combinatorial biocatalysis: a natural approach to drug discovery. , 1998, Trends in biotechnology.

[42]  R. Fernández-Lafuente,et al.  'Interfacial affinity chromatography' of lipases: separation of different fractions by selective adsorption on supports activated with hydrophobic groups. , 1998, Biochimica et biophysica acta.

[43]  Bastida,et al.  A single step purification, immobilization, and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports , 1998, Biotechnology and bioengineering.

[44]  R. Fernández-Lafuente,et al.  Immobilization of lipases by selective adsorption on hydrophobic supports. , 1998, Chemistry and physics of lipids.

[45]  J. Volle,et al.  Easy access to an enantiopure precursor of (+)-goniodiol , 1996 .

[46]  M. Kinoshita,et al.  Factors influencing enantioselectivity of lipase-catalyzed hydrolysis , 1996 .

[47]  George M. Whitesides,et al.  Enzymes in Synthetic Organic Chemistry , 1994 .

[48]  T A Jones,et al.  The sequence, crystal structure determination and refinement of two crystal forms of lipase B from Candida antarctica. , 1994, Structure.

[49]  D. Lawson,et al.  Catalysis at the interface: the anatomy of a conformational change in a triglyceride lipase. , 1992, Biochemistry.

[50]  L. Norskov,et al.  A serine protease triad forms the catalytic centre of a triacylglycerol lipase , 1990, Nature.

[51]  J. Blumbach,et al.  Recent Developments in the Field of β‐Lactam Antibiotics , 1985 .

[52]  D. Koshland,et al.  Reaction of tyrosine residues in proteins with carbodiimide reagents. , 1968, Biochimica et biophysica acta.

[53]  D E Koshland,et al.  A method for the quantitative modification and estimation of carboxylic acid groups in proteins. , 1967, The Journal of biological chemistry.

[54]  L. Sarda,et al.  Action de la lipase pancréatique sur les esters en émulsion , 1958 .