| Title : Protein engineering of stable IsPETase for PET plastic degradation by Premuse - Meng_2021_Int.J.Biol.Macromol_180_667 |
| Author(s) : Meng X , Yang L , Liu H , Li Q , Xu G , Zhang Y , Guan F , Zhang W , Wu N , Tian J |
| Ref : Int J Biol Macromol , 180 :667 , 2021 |
| Abstract : Poly(ethylene terephthalate) (PET) is used widely by human beings, but is very difficult to degrade. Up to now, the PET degradation effect of PETase from Ideonella sakaiensis 201-F6 (IsPETase) variants with low stability and activity was not ideal. In this study, a mutation design tool, Premuse, was developed to integrate the sequence alignment and quantitative selection of the preferred mutations based on natural sequence evolution. Ten single point mutants were selected from 1486 homologous sequences using Premuse, and then two mutations (W159H and F229Y) with improved stability were screened from them. The derived double point mutant, W159H/F229Y, exhibited a strikingly enhanced enzymatic performance. Its T(m) and catalytic efficiency values (k(cat)/K(m)) respectively increased by 10.4 degreesC and 2.0-fold using p-NPP as the substrate compared with wild type. The degradation activity for amorphous PET was increased by almost 40-fold in comparison with wild type at 40 degreesC in 24 h. Additionally, the variant could catalyze biodegradation of PET bottle preform at a mean rate of 23.4 mg(PET)/h/mg(enzyme). This study allowed us to design the mutation more efficiently, and provides a tool for achieving biodegradation of PET pollution under mild natural environments. |
| ESTHER : Meng_2021_Int.J.Biol.Macromol_180_667 |
| PubMedSearch : Meng_2021_Int.J.Biol.Macromol_180_667 |
| PubMedID: 33753197 |
| Gene_locus related to this paper: idesa-peth |
| Gene_locus related to this paper: idesa-peth |
Meng X, Yang L, Liu H, Li Q, Xu G, Zhang Y, Guan F, Zhang W, Wu N, Tian J (2021)
Protein engineering of stable IsPETase for PET plastic degradation by Premuse
Int J Biol Macromol
180 :667
Meng X, Yang L, Liu H, Li Q, Xu G, Zhang Y, Guan F, Zhang W, Wu N, Tian J (2021)
Int J Biol Macromol
180 :667