@article{demonceaux2023enzymatic,

title = {Enzymatic synthesis, characterization and molecular docking of a new functionalized polyphenol: Resveratrol-3, 4’-⍺-diglucoside},

author = {Marie Demonceaux and Marine Goux and Lucia Emanueli Schimith and Michele Goulart Dos Santos and Johann Hendrickx and Bernard Offmann and Corinne André-Miral},

url = {https://www.sciencedirect.com/science/article/pii/S2211715623001959},

doi = {10.1016/j.rechem.2023.100956},

year  = {2023},

date = {2023-05-16},

urldate = {2023-05-16},

journal = {Results in Chemistry},

pages = {100956},

publisher = {Elsevier},

abstract = {Transglucosylation of resveratrol by the Q345F variant of sucrose phosphorylase from Bifidobacterium adolescentis (BaSP) was extensively studied during the last decade. Indeed, Q345F is able to catalyze the synthesis of resveratrol-3-O-⍺-D-glucoside (RES-3) with yield up to 97% using a cost-effective glucosyl donor, sucrose (Kraus et al., Chemical Communications, 53(90), 12182–12184 (2017)). Despite the fact that two further products were detectable in low amounts after glucoside synthesis, they were never identified. Here, we isolated and fully characterized one of those two minor products: resveratrol-3,4′-O-⍺-D-diglucoside (RES-3,4′). This original compound had never been described before. Using bioinformatics models, we successfully explained the formation of this diglucosylated product. Indeed, with RES-3 as acceptor substrate, Q345F is able to transfer a glucosyl moiety in position 4′-OH, what had been reported as impossible in the literature. The low yield observed is due to the steric hindrance into the catalytic site between RES-3 and residues Tyr132 and Tyr344. Nevertheless, the substrate orientation in the active site is favored by stabilizing interactions. Ring A of RES-3 bearing the diol moiety is stabilized by hydrogen bonds with residues Asp50, Arg135, Asn347 and Arg399. Hydroxyl group OH-4′ shares hydrogen bonds with the catalytic residues Asp192 and Glu232. Multiple hydrophobic contacts complete the stabilization of the substrate to favor the glucosylation at position 4′. Understanding of the mechanisms allowing the glucosylation at position 4′ of resveratrol will help the development of enzymatic tools to target and control the enzymatic synthesis of original ⍺-glucosylated polyphenols with high added value and better biodisponibility.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Goux2023.04.11.536264,

title = {Sucrose phosphorylase from Alteromonas mediterranea: structural insight into the regioselective α-glucosylation of (+)-catechin},

author = {Marine Goux and Marie Demonceaux and Johann Hendrickx and Claude Solleux and Emilie Lormeau and Folmer Fredslund and David Tezé and Bernard Offmann and Corinne André-Miral},

url = {https://www.biorxiv.org/content/early/2023/04/11/2023.04.11.536264},

doi = {10.1101/2023.04.11.536264},

year  = {2023},

date = {2023-04-11},

urldate = {2023-04-11},

journal = {bioRxiv},

publisher = {Cold Spring Harbor Laboratory},

abstract = {Flavonoids glycosylation at different positions is paramount to solubility and modulation of bioactivities. Sucrose phosphorylases, through transglycosylation reactions, are interesting enzymes that can transfer glucose from sucrose, the donor substrate, onto polyphenols to form glycoconjugates. Here, we report for the first time the structural and enzymatic properties of sucrose phosphorylase from the marine bacteria Alteromonas mediterranea (AmSP). We characterized and investigated the transglucosylation capacity of two new variants of the enzyme on (+)-catechin and their propensity to catalyse its regioselective glucosylation. AmSP-Q353F and AmSP-P140D were shown to catalyse the regiospecific glucosylation of (+)-catechin using sucrose as donor substrate. While AmSP-WT was devoid of synthetic activity, each of its two single mutant provided high yields of specific regioisomers: 89% of (+)-catechin-4'-O-α-D-glucopyranoside (CAT-4’) for AmSP-P140D and 92% of (+)-catechin-3'-O-α-D-glucopyranoside (CAT-3’) for AmSP-Q353F. The novel compound CAT-4’ was fully characterized by NMR and mass spectrometry. We used molecular docking simulations on structural models of the glucosyl-enzyme intermediate to explain this regioselectivity. We showed that AmSP-P140D preferentially binds (+)-catechin in a mode that favours glucosylation on its hydroxyl group in position 4’ (OH-4’) while the binding mode of the flavonoid in AmSP-Q353F favoured glucosylation on its hydroxyl group in position 3’ (OH-3’).Competing Interest StatementThe authors have declared no competing interest.},

keywords = {},

pubstate = {forthcoming},

tppubtype = {article}

}

@article{demonceaux2023regioselective,

title = {Regioselective glucosylation of (+)-catechin using a new variant of sucrose phosphorylase from Bifidobacterium adolescentis},

author = {Marie Demonceaux and Marine Goux and Johann Hendrickx and Claude Solleux and Frédéric Cadet and Émilie Lormeau and Bernard Offmann and Corinne André-Miral},

doi = {10.1039/D3OB00191A},

year  = {2023},

date = {2023-02-22},

urldate = {2023-02-22},

journal = {Organic & Biomolecular Chemistry},

volume = {21},

number = {11},

pages = {2307--2311},

publisher = {Royal Society of Chemistry},

abstract = {Mutation Q345F in sucrose phosphorylase from Bifidobacterium adolescentis (BaSP) has shown to allow efficient (+)-catechin glucosylation yielding a regioisomeric mixture: (+)-catechin-3′-O-α-D-glucopyranoside, (+)-catechin-5-O-α-D-glucopyranoside and (+)-catechin-3′,5-O-α-D-diglucopyranoside with a ratio of 51 : 25 : 24. Here, we efficiently increased the control of (+)-catechin glucosylation regioselectivity with a new variant Q345F/P134D. The same products were obtained with a ratio of 82 : 9 : 9. Thanks to bioinformatics models, we successfully explained the glucosylation favoured at the OH-3′ position due to the mutation P134D.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Goux2016,

title = {In vivo phosphorylation of a peptide tag for protein purification},

author = {Marine Goux and Amina Fateh and Alain Defontaine and Mathieu Cinier and Charles Tellier},

url = {https://doi.org/10.1007/s10529-016-2040-4},

doi = {10.1007/s10529-016-2040-4},

issn = {1573-6776},

year  = {2016},

date = {2016-01-01},

journal = {Biotechnology Letters},

volume = {38},

number = {5},

pages = {767--772},

abstract = {To design a new system for the in vivo phosphorylation of proteins in Escherichia coli using the co-expression of the α-subunit of casein kinase II (CKIIα) and a target protein, (Nanofitin) fused with a phosphorylatable tag.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}