Boyd ES

References (1)

Title : Sourcing thermotolerant poly(ethylene terephthalate) hydrolase scaffolds from natural diversity - Erickson_2022_Nat.Commun_13_7850
Author(s) : Erickson E , Gado JE , Avilan L , Bratti F , Brizendine RK , Cox PA , Gill R , Graham R , Kim DJ , Konig G , Michener WE , Poudel S , Ramirez KJ , Shakespeare TJ , Zahn M , Boyd ES , Payne CM , Dubois JL , Pickford AR , Beckham GT , McGeehan JE
Ref : Nat Commun , 13 :7850 , 2022
Abstract : Enzymatic deconstruction of poly(ethylene terephthalate) (PET) is under intense investigation, given the ability of hydrolase enzymes to depolymerize PET to its constituent monomers near the polymer glass transition temperature. To date, reported PET hydrolases have been sourced from a relatively narrow sequence space. Here, we identify additional PET-active biocatalysts from natural diversity by using bioinformatics and machine learning to mine 74 putative thermotolerant PET hydrolases. We successfully express, purify, and assay 51 enzymes from seven distinct phylogenetic groups; observing PET hydrolysis activity on amorphous PET film from 37 enzymes in reactions spanning pH from 4.5-9.0 and temperatures from 30-70 degreesC. We conduct PET hydrolysis time-course reactions with the best-performing enzymes, where we observe differences in substrate selectivity as function of PET morphology. We employed X-ray crystallography and AlphaFold to examine the enzyme architectures of all 74 candidates, revealing protein folds and accessory domains not previously associated with PET deconstruction. Overall, this study expands the number and diversity of thermotolerant scaffolds for enzymatic PET deconstruction.
ESTHER : Erickson_2022_Nat.Commun_13_7850
PubMedSearch : Erickson_2022_Nat.Commun_13_7850
PubMedID: 36543766
Gene_locus related to this paper: 9arch-PETcan211 , 9cren-PETcan204 , 9cyan-305pEE028 , 9bact-102Pee006 , 9chlr-7QJM202 , 9bact-a0a656d8b6 , 9actn-a0a1t4kk94 , 9burk-PET11 , 9bact-c3ryl0 , thecs-711Erick , 9actn-RII04304 , 9actn-h6wx58 , thecd-d1a9g5 , thecd-d1a2h1 , 9acto-d4q9n1 , 9acto-f7ix06 , 9gamm-a0a3l8bw54 , 9actn-a0a0n0my27 , 9burk-a0a1e4lw26 , 9actn-Alr407 , 9gamm-a0a3l8bdt3 , 9gamm-a0a2k9lit3 , 9bact-g9by57 , bacsu-pnbae , thefu-q6a0i4 , 9actn-a0a0n0ney5 , 9pseu-a0a1i6nu60 , thefu-q6a0i3 , 9actn-a0a147kjy8 , 9actn-e9upm2