- Reshaping the active pocket of esterase Est816 for resolution of economically important racemates
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Bacterial esterases are potential biocatalysts for the production of optically pure compounds. However, the substrate promiscuity and chiral selectivity of esterases usually have a negative correlation, which limits their commercial value. Herein, an efficient and versatile esterase (Est816) was identified as a promising catalyst for the hydrolysis of a wide range of economically important substrates with low enantioselectivity. We rationally designed several variants with up to 11-fold increased catalytic efficiency towards ethyl 2-arylpropionates, mostly retaining the initial substrate scope and enantioselectivity. These variants provided a dramatic increase in efficiency for biocatalytic applications. Based on the best variant Est816-M1, several variants with higher or inverted enantioselectivity were designed through careful analysis of the structural information and molecular docking. Two stereoselectively complementary mutants, Est816-M3 and Est816-M4, successfully overcame and even reversed the low enantioselectivity, and several 2-arylpropionic acid derivatives with highEvalues were obtained. Our results offer potential industrial biocatalysts for the preparation of structurally diverse chiral carboxylic acids and further lay the foundation for improving the catalytic efficiency and enantioselectivity of esterases.
- Fan, Xinjiong,Fu, Yao,Liu, Xiaolong,Zhao, Meng
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p. 6126 - 6133
(2021/09/28)
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- Efficient resolution of profen ethyl ester racemates by engineered Yarrowia lipolytica Lip2p lipase
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Enzyme-catalyzed enantiomer discrimination is still a great challenge for the development of industrial pharmaceutical processes. For the resolution of ibuprofen, naproxen and ketoprofen racemates, three major anti-inflammatory drugs, only lipases from Candida rugosa present a high selectivity if solvent and surfactant use is discarded. However, their catalytic activities are too low. In the present work, we demonstrate that the lipase Lip2p from the yeast Yarrowia lipolytica has a higher catalytic activity than C. rugosa lipases to hydrolyze the ethyl esters of ibuprofen, naproxen and ketoprofen, but its selectivity is not sufficient [E?=?52 (S); 11 (S) and 1.5 (R) respectively]. The enantioselectivity was further improved by site-directed mutagenesis, targeted at the substrate binding site and guided by molecular modelling studies. By investigating the binding modes of the (R)- and (S)-enantiomers in the active site, two amino acid residues located in the hydrophobic substrate binding site of the lipase, namely residues 232 and 235, were identified as crucial for enantiomer discrimination and enzyme activity. The (S) enantioselectivity of Lip2p towards ethyl ibuprofen esters was rendered infinite (E???300) by replacing V232 by an A or C residue. Substitution of V235 by C, M, S, or T amino acids led to a great increase in the (S)-enantioselectivity (E???300) towards naproxen ethyl ester. Finally, the variant V232F enabled the efficient kinetic resolution of ethyl ketoprofen ester enantiomers [(R)-enantiopreference; E???300]. In addition to the increase in selectivity, a remarkable increase in velocity by 2.6, 2.7 and 2.5?times, respectively, was found for ibuprofen, naproxen and ketoprofen ethyl esters.
- Gérard, Doriane,Guéroult, Marc,Casas-Godoy, Leticia,Condoret, Jean-Stéphane,André, Isabelle,Marty, Alain,Duquesne, Sophie
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p. 433 - 441
(2017/03/24)
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- Fabrication of organogels achieved by prodrug-based organogelators of ketoprofen
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The treatment strategy of curing diseases using prodrugs of an anti-inflammatory drug is widespread. In the present study, we report on the synthesis of prodrugs of ketoprofen, consisting of a derivatization of ketoprofen and long hydrocarbon chain of fat
- Mahire, Rahul R.,Agrawal, Deepika S.,Patil, Devanand K.,More, Dhananjay H.
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p. 33286 - 33291
(2014/08/18)
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- Comparison of the enzymatic activity of commercial and semipurified lipase of Candida cylindracea in the hydrolysis of the esters of (R,S) 2-aryl propionic acids
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A semipurified lipase of Candida cylindradea (LS) - easily obtained from commercial crude lipase (LC) - is used in the enantioselective hydrolysis of (R,S) 2-arylpropionates. The semipurification treatment diminishes the lipase activity more than the esterase activity. The addition of lactose (24 h) increases both activities. LS is more active than LC - at the same amount of protein - in the hydrolysis of (R,S) 2-aryl propionates. This semipurification showed a remarkable improvement in yield in the enantioespecific hydrolysis of these esters.
- Hernaiz, Maria J.,Sanchez-Montero, Jose M.,Sinisterra, Jose V.
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p. 10749 - 10760
(2007/10/02)
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- A NEW METHOD OF SYNTHESIS OF α-ARYLPROPIONIC ACIDS INVOLVING COPPER(I) HOMOGENEOUS CATALYSIS
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The synthesis of α-arylpropionic acids involving, as first step, the facile coupling af an aryl bromide and diethyl malonate in the presence of Cu(I) bromide is described.The limits of application are discussed.
- Ugo, Renato,Nardi, Paola,Psaro, Rinaldo,Roberto, Dominique
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p. 511 - 514
(2007/10/02)
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- Aminoazole derivatives and their production and use
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A compound of the formula: STR1 wherein A is the group of the formula: wherein Ar1 is a phenyl or thienyl group which may be optionally substituted with at least one of the same or different halogen atom; Ar2 is a phenylene or thienylene group which may be optionally substituted with at least one of the same or different halogen atom; D is a divalent radical selected from the group consisting of >C=N--OR4 [wherein R4 is a hydrogen atom or lower alkyl group], >C=O, STR2 >CHOH, >NH radical, or single bond, STR3 wherein R5 is a lower alkoxy or a phenyl group which may be optionally substituted with at least one of the same or different halogen atom; E is a methine group or a nitrogen atom; F is a vinylene group or an oxygen atom, STR4 wherein R6 is a lower alkoxy group; R7 is a lower alkyl group; R8 is a benzoyl group which may be optionally substituted with at least one of the same or different halogen atom, B is a divalent azole group; R1 is a hydrogen atom or a lower alkyl group; R2 is a hydrogen atom, lower alkyl, aryl-lower alkyl, or the group of the formula: wherein R9 is a hydrogen atom, lower alkyl, halo-lower alkyl, amino-lower alkyl, aryl or aryl-lower alkyl group or the group of the formula: STR5 wherein R10 is a hydrogen atom or lower alkoxy group; R11 is a hydrogen atom, lower alkyl, lower alkenyl, lower cycloalkyl, aryl-lower alkyl, aryl or aroyl group; or the group of the formula: --NR10 R11 is a 5-, 6- or 7-membered saturated heterocyclic ring; or the group of the formula: wherein R12 is a lower alkyl or polyhalo-lower alkyl group; G is a divalent group selected from the group consisting of >C=O, >C=S, >(C=O)2 or >SO2 radical; or the group of the formula: --NR1 R2 is a 5-, 6- or 7-membered saturated heterocyclic ring: R3 is a hydrogen atom or lower alkyl group, or its acid addition salts, which is useful for immunomodulator.
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- A NOVEL SYNTHESIS OF KETOPROFEN, IMPORTANT NON-STEROIDAL ANTIINFLAMMATORY AGENT
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The α-propionic acid moiety in ketoprofen is attached through Wittig Horner reaction of ethylphosphono propionate on 3-benzyl cyclohexanone, prepared by G. reaction of benzylmagnesium chloride with 2-cyclohexenone (in presence of cuprous chloride).
- Mitra, R. B.,Joshi, Vijaya S.
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p. 2259 - 2266
(2007/10/02)
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