Huguenin-Dezot N

References (3)

Title : Discovery and Genetic Code Expansion of a Polyethylene Terephthalate (PET) Hydrolase from the Human Saliva Metagenome for the Degradation and Bio-Functionalization of PET - Eiamthong_2022_Angew.Chem.Int.Ed.Engl_15_e202203061
Author(s) : Eiamthong B , Meesawat P , Wongsatit T , Jitdee J , Sangsri R , Patchsung M , Aphicho K , Suraritdechachai S , Huguenin-Dezot N , Tang S , Suginta W , Paosawatyanyong B , Babu MM , Chin JW , Pakotiprapha D , Bhanthumnavin W , Uttamapinant C
Ref : Angew Chem Int Ed Engl , :e202203061 , 2022
Abstract : We report a bioinformatic workflow and subsequent discovery of a new polyethylene terephthalate (PET) hydrolase, which we named MG8, from the human saliva metagenome. MG8 has robust PET plastic degradation activities under different temperature and salinity conditions, outperforming several naturally occurring and engineered hydrolases in degrading PET. Moreover, we genetically encoded 2,3-diaminopropionic acid (DAP) in place of the catalytic serine residue of MG8, thereby converting a PET hydrolase into a covalent binder for bio-functionalization of PET. We show that MG8(DAP), in conjunction with a split green fluorescent protein system, can be used to attach protein cargos to PET as well as other polyester plastics. The discovery of a highly active PET hydrolase from the human metagenome-currently an underexplored resource for industrial enzyme discovery-as well as the repurposing of such an enzyme into a plastic functionalization tool, should facilitate ongoing efforts to degrade and maximize reusability of PET.
ESTHER : Eiamthong_2022_Angew.Chem.Int.Ed.Engl_15_e202203061
PubMedSearch : Eiamthong_2022_Angew.Chem.Int.Ed.Engl_15_e202203061
PubMedID: 35656865
Gene_locus related to this paper: 9gamm-PETaseMG1 , 9gamm-PETaseMG2 , 9gamm-PETaseMG3 , 9gamm-PETaseMG4 , 9gamm-PETaseMG5 , 9gamm-PETaseMG6 , 9gamm-PETaseMG7 , 9gamm-PETaseMG8 , 9pseu-PETaseMG9 , 9actn-PETaseMG10

Title : Mechanism-based traps enable protease and hydrolase substrate discovery - Tang_2022_Nature_602_701
Author(s) : Tang S , Beattie AT , Kafkova L , Petris G , Huguenin-Dezot N , Fiedler M , Freeman M , Chin JW
Ref : Nature , 602 :701 , 2022
Abstract : Hydrolase enzymes, including proteases, are encoded by 2-3% of the genes in the human genome and 14% of these enzymes are active drug targets(1). However, the activities and substrate specificities of many proteases-especially those embedded in membranes-and other hydrolases remain unknown. Here we report a strategy for creating mechanism-based, light-activated protease and hydrolase substrate traps in complex mixtures and live mammalian cells. The traps capture substrates of hydrolases, which normally use a serine or cysteine nucleophile. Replacing the catalytic nucleophile with genetically encoded 2,3-diaminopropionic acid allows the first step reaction to form an acyl-enzyme intermediate in which a substrate fragment is covalently linked to the enzyme through a stable amide bond(2); this enables stringent purification and identification of substrates. We identify new substrates for proteases, including an intramembrane mammalian rhomboid protease RHBDL4 (refs. (3,4)). We demonstrate that RHBDL4 can shed luminal fragments of endoplasmic reticulum-resident type I transmembrane proteins to the extracellular space, as well as promoting non-canonical secretion of endogenous soluble endoplasmic reticulum-resident chaperones. We also discover that the putative serine hydrolase retinoblastoma binding protein 9 (ref. (5)) is an aminopeptidase with a preference for removing aromatic amino acids in human cells. Our results exemplify a powerful paradigm for identifying the substrates and activities of hydrolase enzymes.
ESTHER : Tang_2022_Nature_602_701
PubMedSearch : Tang_2022_Nature_602_701
PubMedID: 35173328
Gene_locus related to this paper: human-RBBP9

Title : Trapping biosynthetic acyl-enzyme intermediates with encoded 2,3-diaminopropionic acid - Huguenin-Dezot_2019_Nature_565_112
Author(s) : Huguenin-Dezot N , Alonzo DA , Heberlig GW , Mahesh M , Nguyen DP , Dornan MH , Boddy CN , Schmeing TM , Chin JW
Ref : Nature , 565 :112 , 2019
Abstract : Many enzymes catalyse reactions that proceed through covalent acyl-enzyme (ester or thioester) intermediates(1). These enzymes include serine hydrolases(2,3) (encoded by one per cent of human genes, and including serine proteases and thioesterases), cysteine proteases (including caspases), and many components of the ubiquitination machinery(4,5). Their important acyl-enzyme intermediates are unstable, commonly having half-lives of minutes to hours(6). In some cases, acyl-enzyme complexes can be stabilized using substrate analogues or active-site mutations but, although these approaches can provide valuable insight(7-10), they often result in complexes that are substantially non-native. Here we develop a strategy for incorporating 2,3-diaminopropionic acid (DAP) into recombinant proteins, via expansion of the genetic code(11). We show that replacing catalytic cysteine or serine residues of enzymes with DAP permits their first-step reaction with native substrates, allowing the efficient capture of acyl-enzyme complexes that are linked through a stable amide bond. For one of these enzymes, the thioesterase domain of valinomycin synthetase(12), we elucidate the biosynthetic pathway by which it progressively oligomerizes tetradepsipeptidyl substrates to a dodecadepsipeptidyl intermediate, which it then cyclizes to produce valinomycin. By trapping the first and last acyl-thioesterase intermediates in the catalytic cycle as DAP conjugates, we provide structural insight into how conformational changes in thioesterase domains of such nonribosomal peptide synthetases control the oligomerization and cyclization of linear substrates. The encoding of DAP will facilitate the characterization of diverse acyl-enzyme complexes, and may be extended to capturing the native substrates of transiently acylated proteins of unknown function.
ESTHER : Huguenin-Dezot_2019_Nature_565_112
PubMedSearch : Huguenin-Dezot_2019_Nature_565_112
PubMedID: 30542153
Gene_locus related to this paper: 9acto-q1psf3