Targeted Genome Mining Facilitates the Discovery of a Promiscuous, Hyperthermostable Amidase from Thermovenabulum gondwanense with Notable Nylon-Degrading Capacity

Plastics are ubiquitous in our ecosystems, and microplastic accumulation in the environment is an emerging global health concern. Since available recycling technologies are not economically competitive with primary plastic production, global use is expected to reach 1231 megatons by 2060, with 493 megatons leeching into the environment each year. To identify new nylon-recycling biotechnologies, targeted genome mining was used to identify thermostable enzymes capable of degrading polyamides. Here, we describe the characterization of a novel protein sourced from Thermovenabulum gondwanense: TvgC. TvgC is extremely stable, exhibiting a melting temperature of 93 °C and no detectable losses in hydrolytic activity after one week at 60 °C. While nylonases primarily process nylon-6, TvgC catalysed the degradation of both nylon-6 and nylon-6,6 films, exhibiting marked preference for nylon-6,6, which is considered more difficult to degrade. Finally, conversion experiments demonstrate that TvgC achieves a 1.2 wt% conversion of nylon-6 film, outcompeting the most highly engineered nylonases. This novel hyperthermostable protein represents an excellent starting point for future engineering of increasingly efficient nylonases.