Optimisation of stabilised Carboxylesterase NP for enantioselective hydrolysis of naproxen methyl ester

Steenkamp, Lucia H; Brady, Dean

doi:10.1016/j.procbio.2008.09.001

Cited by 20 publications

(15 citation statements)

References 26 publications

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“…The synthesis of pharmaceutical intermediates is increasingly becoming the function of biocatalytic processes due to their high specificity (Pollard and Woodley, 2007); multi-enzyme reactions are becoming more viable (Findrik and Vasić-Rački, 2007); and enzymes are being shown capable of converting poorly soluble substrates (Wolff et al, 1997;Steenkamp and Brady, 2008).…”

Section: Effect Of Combined Factors On Reaction Performancementioning

confidence: 99%

Defining a process operating window for the synthesis of 5-methyluridine by transglycosylation of guanosine and thymine

Gordon

Visser

Brady

et al. 2011

Journal of Biotechnology

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Section: Effect Of Combined Factors On Reaction Performancementioning

confidence: 99%

Defining a process operating window for the synthesis of 5-methyluridine by transglycosylation of guanosine and thymine

Gordon

Visser

Brady

et al. 2011

Journal of Biotechnology

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“…Bacteria belonging to the genus Bacillus are known producers of stereoselective carboxylesterases . Lipolytic and nonlipolytic esterases from Bacillus subtilis , Bacillus coagulans , Bacillus amyloliquefaciens , Bacillus stearothermophilus , and generic Bacillus sp. have been identified as excellent biocatalysts for the stereoselective hydrolysis of chiral and prochiral esters.…”

Section: Introductionmentioning

confidence: 99%

A stereospecific carboxyl esterase from Bacillus coagulans hosting nonlipase activity within a lipase‐like fold

et al. 2018

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“…NP has a product ee of 96% based on the substrate S-naproxenmethylester ( Fig. 1) and 99% on S-naproxenethylester (E N 200) [3,4]. In contrast, the enzyme's enantioselectivity towards 1,2-O-isopropylideneglycerol esters (IPG esters) ( Fig.…”

Section: Introductionmentioning

confidence: 99%

Crystal structures of two Bacillus carboxylesterases with different enantioselectivities

Rozeboom

Godinho

Nardini

et al. 2014

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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Naproxen esterase (NP) from Bacillus subtilis Thai I-8 is a carboxylesterase that catalyzes the enantioselective hydrolysis of naproxenmethylester to produce S-naproxen (E N 200). It is a homolog of CesA (98% sequence identity) and CesB (64% identity), both produced by B. subtilis strain 168. CesB can be used for the enantioselective hydrolysis of 1,2-O-isopropylideneglycerol (solketal) esters (E N 200 for IPG-caprylate). Crystal structures of NP and CesB, determined to a resolution of 1.75 Å and 2.04 Å, respectively, showed that both proteins have a canonical α/β hydrolase fold with an extra N-terminal helix stabilizing the cap subdomain. The active site in both enzymes is located in a deep hydrophobic groove and includes the catalytic triad residues Ser130, His274, and Glu245. A product analog, presumably 2-(2-hydroxyethoxy)acetic acid, was bound in the NP active site. The enzymes have different enantioselectivities, which previously were shown to result from only a few amino acid substitutions in the cap domain. Modeling of a substrate in the active site of NP allowed explaining the different enantioselectivities. In addition, Ala156 may be a determinant of enantioselectivity as well, since its side chain appears to interfere with the binding of certain R-enantiomers in the active site of NP. However, the exchange route for substrate and product between the active site and the solvent is not obvious from the structures. Flexibility of the cap domain might facilitate such exchange. Interestingly, both carboxylesterases show higher structural similarity to meta-cleavage compound (MCP) hydrolases than to other α/β hydrolase fold esterases.

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Optimisation of stabilised Carboxylesterase NP for enantioselective hydrolysis of naproxen methyl ester

Cited by 20 publications

References 26 publications

Defining a process operating window for the synthesis of 5-methyluridine by transglycosylation of guanosine and thymine

Defining a process operating window for the synthesis of 5-methyluridine by transglycosylation of guanosine and thymine

A stereospecific carboxyl esterase from Bacillus coagulans hosting nonlipase activity within a lipase‐like fold

Crystal structures of two Bacillus carboxylesterases with different enantioselectivities

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