Diversity of sugar acceptor of glycosyltransferase 1 from Bacillus cereus and its application for glucoside synthesis

Abstract

Glycosyltransferase 1 from Bacillus cereus (BcGT1) catalyzes the transfer of a glucosyl moiety from uridine diphosphate glucose (UDP-glucose) to various acceptors; it was expressed and characterized. The specificity of acceptors was found to be broad: more than 20 compounds classified into O-, S-, and N-linkage glucosides can be prepared with BcGT1 catalysis. Based on this work, we conclude that the corresponding acceptors of these compounds must possess the following features: (1) the acceptors must contain at least one aromatic or fused-aromatic or heteroaromatic ring; (2) the reactive hydroxyl or sulfhydryl or amino group can attach either on the aromatic ring or on its aliphatic side chain; and (3) the acceptors can be a primary, secondary, or even a tertiary amine. Four representative acceptors-fluorescein methyl ester, 17-β-estradiol, 7-mercapto-4-methylcoumarin, and 6-benzylaminopurine-were chosen as a candidate acceptor for O-, S-, and N-glucosidation, respectively. These enzymatic products were purified and the structures were confirmed with mass and NMR spectra. As all isolated glucosides are β-anomers, BcGT1 is confirmed to be an inverting enzyme. This study not only demonstrates the substrate promiscuity of BcGT1 but also showed the great application prospect of this enzyme in bioconversion of valuable bioactive molecules.

Thioglycoligase derived from fungal GH3 β-xylosidase is a multi-glycoligase with broad acceptor tolerance

“…These enzymes have traditionally sparked attention for being the biocatalysts responsible for synthesizing most of the natural glyconjugates. In the last decade, efforts have been made to increase their acceptor range, successfully reporting glycosyltransferases able to form O-, C-, S-, and/or N-glycosidic bonds [72][73][74][75] . However, the application of glycosyltransferases is still hampered by the high cost of the activated sugars required as glycosylation donors.…”

Selective β-Mono-Glycosylation of a C15-Hydroxylated Metabolite of the Agricultural Herbicide Cinmethylin Using Leloir Glycosyltransferases

“…The following constructs were used considering the literature on the respective enzyme. Bc GT1 (origin: B. cereus ; GenBank accession number: KT821092; no tag); 37 OleD wildtype (origin: S. antibioticus ; GenBank accession number: DQ195536.2; also referred to as UGT102A2; N -terminal His tag); 38 , 39 OleD triple variant ASP (A242V/S132F/P167T; N -terminal His tag); 38 , 39 UGT71E5 (origin: C. tinctorius ; GenBank accession number: KX610759.1; N -terminal His tag); 32 UGT1A9 (origin: Homo sapiens; GenBank accession number: AF056188.1; N -terminal maltose binding protein; C-terminal His tag); 46 arbutin synthase (origin: R. serpentina ; GenBank accession number: CAC35167.1; N -terminal Strep tag); 33 , 35 UGT71A15 (origin: M. domestica ; GenBank accession number: DQ103712; N -terminal Strep tag), 34 and UGT708A6 (origin: Z. mays ; GenBank accession number: ACF81582.1; N -terminal Strep tag) 33 , 47 were obtained as described recently.…”

Insight on Mercapto-Coumarins: Synthesis and Reactivity

“…It is also indicated as a suitable matrix for the analysis of small molecular compounds [ 40 ]. The other described incorporation of this highly valuable scaffold is as a reporter molecule [ 41 ], reporters for thiol interactions at the nanoparticle surface [ 42 ], fluorescent probe [ 43 , 44 , 45 ], sugar acceptor [ 46 ], Raman reporter [ 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 ], an excellent substrate for fluorescence spectroscopy [ 57 ], photodimerizable and healable reactant [ 58 ], fluorescent dye [ 59 , 60 ], and probe molecule on gold-coated silicon nanowires [ 61 ].…”