Pennacchio_2015_Extremophiles_19_1001

Reference

Title : Enlarging the substrate portfolio of the thermophilic esterase EST2 from Alicyclobacillus acidocaldarius - Pennacchio_2015_Extremophiles_19_1001
Author(s) : Pennacchio A , Mandrich L , Manco G , Trincone A
Ref : Extremophiles , 19 :1001 , 2015
Abstract : The enzymatic regioselective hydrolysis of (a) acetylated mono- to tetrasaccharides of different nature, (b) of acetylated aryl glycosides and (c) of different acetylated nucleosides was studied enlarging the portfolio of substrates that can be employed by the thermophilic esterase EST2 from Alicyclobacillus acidocaldarius. The reactions were optimised to the extent that the amount of enzyme needed was lowered of two orders of magnitude with respect to the previously reported reactions, namely from 4000 to 40 U of enzyme per reaction. New additional solvents were screened and dramatic changes in regioselectivity were observed depending on the amount and type of solvent used. For example, in the presence of 10 % DMF, only two alpha-D-glucose products 6-OH and 4,6-OH (in a 76:24 ratio) were detected, whereas with 25 % DMF, at least four products of similar amount were observed. This versatility adds specific value to the biocatalyst making possible the design of biocatalytic reactions with different hydrophobic ester substrates. As an additional remarkable example, EST2 catalysed with a good yield and high regioselectivity the hydrolysis of p-nitrophenyl beta-D-xylopyranoside triacetate producing only the monoacetylated derivative with acetyl group in 3-O-position, in 2 min. The results with nucleosides as substrates are particularly interesting. The peracetates of 3',5'-di-O-acetylthymidine are converted almost quantitatively (95 %) to the monoacetylated derivative possessing free secondary OH; this regioselectivity is complementary to hydrolysis/alcoholysis reactions catalysed by CAL-B lipase or to other microbial hydrolytic biocatalysts, generally giving products with free primary OH groups. A docking analysis was undertaken with all analysed substrates suggesting a structural interpretation of the results. In most of cases, the best pose of the selected substrate was in line with the observed regioselectivity.
ESTHER : Pennacchio_2015_Extremophiles_19_1001
PubMedSearch : Pennacchio_2015_Extremophiles_19_1001
PubMedID: 26216109
Gene_locus related to this paper: aliac-est2

Related information

Gene_locus related to this paper: aliac-est2

Citations formats

Pennacchio A, Mandrich L, Manco G, Trincone A (2015)
Enlarging the substrate portfolio of the thermophilic esterase EST2 from Alicyclobacillus acidocaldarius
Extremophiles 19 :1001

Pennacchio A, Mandrich L, Manco G, Trincone A (2015)
Extremophiles 19 :1001