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Enzyme selection, optimization, and production toward biodegradation of post-consumer poly(ethylene terephthalate) at scale.
Soong, Ya-Hue Valerie; Abid, Umer; Chang, Allen C; Ayafor, Christian; Patel, Akanksha; Qin, Jiansong; Xu, Jin; Lawton, Carl; Wong, Hsi-Wu; Sobkowicz, Margaret J; Xie, Dongming.
Afiliación
  • Soong YV; Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Abid U; Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Chang AC; Department of Plastics Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Ayafor C; Energy Engineering Program, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Patel A; Department of Plastics Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Qin J; Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Xu J; Department of Chemistry, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Lawton C; Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Wong HW; Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Sobkowicz MJ; Department of Plastics Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
  • Xie D; Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, Massachusetts, USA.
Biotechnol J ; 18(12): e2300119, 2023 Dec.
Article en En | MEDLINE | ID: mdl-37594123
ABSTRACT
Poly(ethylene terephthalate) (PET) is one of the world's most widely used polyester plastics. Due to its chemical stability, PET is extremely difficult to hydrolyze in a natural environment. Recent discoveries in new polyester hydrolases and breakthroughs in enzyme engineering strategies have inspired enormous research on biorecycling of PET. This study summarizes our research efforts toward large-scale, efficient, and economical biodegradation of post-consumer waste PET, including PET hydrolase selection and optimization, high-yield enzyme production, and high-capacity enzymatic degradation of post-consumer waste PET. First, genes encoding PETase and MHETase from Ideonella sakaiensis and the ICCG variant of leaf-branch compost cutinase (LCCICCG ) were codon-optimized and expressed in Escherichia coli BL21(DE3) for high-yield production. To further lower the enzyme production cost, a pelB leader sequence was fused to LCCICCG so that the enzyme can be secreted into the medium to facilitate recovery. To help bind the enzyme on the hydrophobic surface of PET, a substrate-binding module in a polyhydroxyalkanoate depolymerase from Alcaligenes faecalis (PBM) was fused to the C-terminus of LCCICCG . The resulting four different LCCICCG variants (LCC, PelB-LCC, LCC-PBM, and PelB-LCC-PBM), together with PETase and MHETase, were compared for PET degradation efficiency. A fed-batch fermentation process was developed to produce the target enzymes up to 1.2 g L-1 . Finally, the best enzyme, PelB-LCC, was selected and used for the efficient degradation of 200 g L-1 recycled PET in a well-controlled, stirred-tank reactor. The results will help develop an economical and scalable biorecycling process toward a circular PET economy.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ácidos Ftálicos / Tereftalatos Polietilenos Idioma: En Revista: Biotechnol J Asunto de la revista: BIOTECNOLOGIA Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ácidos Ftálicos / Tereftalatos Polietilenos Idioma: En Revista: Biotechnol J Asunto de la revista: BIOTECNOLOGIA Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos