Molecular rules for chemo- and regio-selectivity of Candida antarctica lipase B in peptide acylation reactions

Ferrari, Florent; Paris, Cédric; Maigret, Bernard; Bidouil, Christelle; Delaunay, Stéphane; Humeau, Catherine; Chevalot, Isabelle

doi:10.1016/j.molcatb.2013.12.007

Cited by 12 publications

(12 citation statements)

References 53 publications

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“…The second one provided information on the proximity between the hydrogen atom of the group becoming acylated and the His224:Nε (distance2). These distance criteria have already been successfully used to explain and even predict the selectivity of acylation reactions (De Oliveira et al, 2009;Ferrari et al, 2014). When the lysine was in the α orientation, the distance 1 was lower than 4Å for 8 out of 26 poses, vs. 7 poses out of 15 in the case of the ε orientation.…”

Section: Flexible Docking Resultsmentioning

confidence: 99%

Molecular rules for selectivity in lipase-catalysed acylation of lysine

Dettori

Jelsch

Guiavarc’h

et al. 2018

Process Biochemistry

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The selectivity of L-lysine acylation by lauric acid catalysed by Candida antarctica lipase B (CALB) was investigated combining experimental and theoretical methodologies. Experiments showed the near-exclusive acylation of lysine ε-amino group; only traces of product resulting from the acylation of lysine α-amino group were observed fleetingly. Molecular modelling simulations were performed aiming to understand the molecular rules for selectivity. Flexible docking simulations combined with structural investigations into lysine/CALB binding modes also suggested the preferential acylation of lysine ε-amino group without, however, excluding the acylation of the lysine α-amino group. Electrostatic interaction energy between lysine and the residues covering the catalytic cavity was calculated in order to understand the discrimination between the two lysine amino groups. The results suggests that the proximity of the carboxylate group hinders the binding of the substrate in configurations enabling the Nacylation. Key interactions with the polar region covering the catalytic triad were identified and a plausible explanation for selectivity was proposed.

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Section: Flexible Docking Resultsmentioning

confidence: 99%

Molecular rules for selectivity in lipase-catalysed acylation of lysine

Dettori

Jelsch

Guiavarc’h

et al. 2018

Process Biochemistry

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“…2). In previous study, CALB was shown to catalyze SYK acylation mainly on the lysine side chain [30]. These results suggested the selective N-acylation on α position for aminoacylases.…”

Section: Research For An Aminoacylase Activity In a Crude Extract Of mentioning

confidence: 72%

“…3). Indeed, analysis of the product highlighted the formation of one mono-acylated molecule and the MS 2 profile showed the presence of the fragments [(KW) This specificity differs from the one of CALB as a previous study showed that CALB catalyzed LQKW acylation mainly on the lysine side chain [30]. It has also to be noticed that no acylation of peptides on N-terminal position occurred with Sm-PVA from S. mobaraensis.…”

Section: Research For An Aminoacylase Activity In a Crude Extract Of mentioning

confidence: 92%

“…Aminoacylase synthetic activities of S. ambofaciens crude extract regarding the acylation of lysine by oleic acid was also investigated. A mass spectrometry based method has previously been developed and validated to determine enzyme regio-selectivity from fragments provided by LC-MS-MS [6,30]. A mass spectrometry based method has previously been developed and validated to determine enzyme regio-selectivity from fragments provided by LC-MS-MS [6,30].…”

Section: Research For An Aminoacylase Activity In a Crude Extract Of mentioning

confidence: 99%

“…The regio-selectivity of the reaction catalyzed by S. ambofaciens crude extract was compared to that obtained with the commonly used CALB [6,7,29]. A mass spectrometry based method has previously been developed and validated to determine enzyme regio-selectivity from fragments provided by LC-MS-MS [6,30]. (Fig.…”

Section: Research For An Aminoacylase Activity In a Crude Extract Of mentioning

confidence: 99%

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An aminoacylase activity from Streptomyces ambofaciens catalyzes the acylation of lysine on α‐position and peptides on N‐terminal position

Dettori

Ferrari

Framboisier

et al. 2018

Engineering in Life Sciences

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The presence of aminoacylase activities was investigated in a crude extract of Streptomyces ambofaciens ATCC23877. First activities catalyzing the hydrolysis of N‐α or ε‐acetyl‐L‐lysine were identified. Furthermore, the acylation of lysine and different peptides was studied and compared with results obtained with lipase B of Candida antarctica (CALB). Different regioselectivities were demonstrated for the two classes of enzymes. CALB was able to catalyze acylation only on the ε‐position whereas the crude extract from S. ambofaciens possessed the rare ability to catalyze the N‐acylation on the α‐position of the lysine or of the amino‐acid in N‐terminal position of peptides. Two genes, SAM23877_1485 and SAM23877_1734, were identified in the genome of Streptomyces ambofaciens ATCC23877 whose products show similarities with the previously identified aminoacylases from Streptomyces mobaraensis. The proteins encoded by these two genes were responsible for the major aminoacylase hydrolytic activities. Furthermore, we show that the hydrolysis of N‐α‐acetyl‐L‐lysine could be attributed to the product of SAM23877_1734 gene.

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