Perez-Garcia_2021_Methods.Enzymol_648_137

Plastics are extensively used in our daily life, but they are also a major pollutant of our biosphere accumulating in both the ocean and the land. In the recent years, few enzymes and microorganisms have been discovered with the ability to degrade even fewer synthetic polymers. Nevertheless, more active species and enzymes need to be discovered and described in order to gain more knowledge about protein adaptation to the degradation of not-naturally-occurring polymers. Within this chapter, we focus on efficient methods to identify novel polyethylene terephthalate-degrading enzymes (PETases) from culturable and non-culturable microorganisms by a combination of sequence- and function-based screening. This protocol can be adapted to discover other plastic hydrolases and in general for other enzymes, for which not many characterized specimens are yet available.