Institute for Research Initiatives, Nara Institute of Science and Technology & Division of Biological Science, Nara Institute of Science and Technology, Ikoma, Nara, Japan.
Kyoto Institute of Technology, Kyoto, Japan.
Methods Enzymol. 2021;648:187-205. doi: 10.1016/bs.mie.2020.12.007. Epub 2021 Jan 9.
Few reports have described the biological degradation or utilization of poly(ethylene terephthalate) (PET) to support microbial growth. We screened environmental samples from a PET bottle recycling site and identified the microbial consortium no. 46, which degraded amorphous PET at ambient temperature; thereafter, we isolated the resident Ideonella sakaiensis 201-F6 strain responsible for the degradation. We further identified two hydrolytic enzymes from I. sakaiensis, PET hydrolase (PETase) and mono(2-hydroxyethyl) terephthalate hydrolase (MHETase), which synergistically converted PET into its monomeric building blocks. Here, we provide original methods of microbial screening and isolation of PET degrading microbe(s). These novel approaches can be adapted for exploring microorganisms that degrade PET and other plastics. Furthermore, our enzyme assay protocols to characterize PETase and MHETase can be applied to evaluate new enzymes that target PET and its hydrolysates.
鲜有报道描述过聚对苯二甲酸乙二醇酯（PET）的生物降解或利用情况，以支持微生物的生长。我们从一个 PET 瓶回收点筛选了环境样本，并鉴定出了能够在环境温度下降解无定形 PET 的微生物混合菌群 46 号，随后我们分离出了负责降解的定居菌解凯氏菌 201-F6。我们进一步从解凯氏菌中鉴定出两种水解酶，即 PET 水解酶（PETase）和单（2-羟乙基）对苯二甲酸水解酶（MHETase），它们协同作用将 PET 转化为其单体构件。在此，我们提供了微生物筛选和分离 PET 降解菌的原始方法。这些新方法可以用于探索能够降解 PET 和其他塑料的微生物。此外，我们用于表征 PETase 和 MHETase 的酶分析方案可用于评估针对 PET 及其水解产物的新酶。
