"Some enzymes work great in laboratory experiments for a few hours, but they lose their activity very quickly and the substrate is not completely degraded," says HZB expert Gert Weber. This is not a problem in the test tube in the laboratory, but it is when used in a large bioreactor. Together with the biotech company Carbios, Uwe Bornscheuer and Gert Weber show how new enzymes for PET degradation can be better compared with each other. "In order to allow upscaling later, many parameters must be within a narrow range even in laboratory experiments. The starting material must be precisely defined and the test protocols must be more standardised in order to better assess the performance of the enzymes and their application on an industrial scale," explains Bornscheuer. The researchers have therefore developed a standardised PET hydrolysis protocol that defines reaction conditions relevant for hydrolysis on a larger scale. In particular, two PET materials were used, firstly a defined PET film and secondly PET granulate from waste bottles, as used by Carbios on a technical scale. They used these materials to test four recently discovered PET-decomposing enzymes: LCC-ICCG, FAST-PETase, HotPETase and PES-H1L92F/Q94Y.
When experimenting under this protocol, they found that two of these enzymes, FAST-PETase and HotPETase, were less suitable for large-scale use. This is mainly due to their relatively low depolymerisation rates. PES-H1L92F/Q94Y performed better. The fourth candidate, LCC-ICCG, outperformed the other enzymes by far: LCC-ICCG converts 98% of PET into the monomeric products terephthalic acid (TPA) and ethylene glycol (EG) in 24 hours. "In addition, we were able to reduce the amount of enzyme required for LCC-ICCG by a factor of 3 and the reaction temperature from 72 to 68 °C, which makes the use of this enzyme more economical," says Bornscheuer.