Singh_2017_Protein.Eng.Des.Sel_30_559

Reference

Title : Role of an N-terminal extension in stability and catalytic activity of a hyperthermostable alpha\/beta hydrolase fold esterase - Singh_2017_Protein.Eng.Des.Sel_30_559
Author(s) : Singh MK , Shivakumaraswamy S , Gummadi SN , Manoj N
Ref : Protein Engineering Des Sel , 30 :559 , 2017
Abstract :

The carbohydrate esterase family 7 (CE7) enzymes catalyze the deacetylation of acetyl esters of a broad range of alcohols and is unique in its activity towards cephalosporin C. The CE7 fold contains a conserved N-terminal extension that distinguishes it from the canonical alpha/beta hydrolase fold. The hexameric quaternary structure indicates that the N-terminus may affect activity and specificity by controlling access of substrates to the buried active sites via an entrance tunnel. In this context, we characterized the catalytic parameters, conformation and thermal stability of two truncation variants lacking four and ten residues of the N-terminal region of the hyperthermostable Thermotoga maritima CE7 acetyl esterase (TmAcE). The truncations did not affect the secondary structure or the fold but modulated the oligomerization dynamics. A modest increase was observed in substrate specificity for acetylated xylose compared with acetylated glucose. A drastic reduction of ~30-40 degrees C in the optimum temperature for activity of the variants indicated lower thermal stability. The loss of hyperthermostability appears to be an indirect effect associated with an increase in the conformational flexibility of an otherwise rigid neighboring loop containing a catalytic triad residue. The results suggest that the N-terminal extension was evolutionarily selected to preserve the stability of the enzyme.

PubMedSearch : Singh_2017_Protein.Eng.Des.Sel_30_559
PubMedID: 28967962
Gene_locus related to this paper: thema-TM0077

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Citations formats

Singh MK, Shivakumaraswamy S, Gummadi SN, Manoj N (2017)
Role of an N-terminal extension in stability and catalytic activity of a hyperthermostable alpha\/beta hydrolase fold esterase
Protein Engineering Des Sel 30 :559

Singh MK, Shivakumaraswamy S, Gummadi SN, Manoj N (2017)
Protein Engineering Des Sel 30 :559