期刊
FRONTIERS IN MICROBIOLOGY
卷 11, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fmicb.2020.00114
关键词
Pseudomonas aestusnigri; marine bacteria; polyester degradation; polyethylene terephthalate; PET; crystal structure
类别
资金
- European Union's Horizon 2020 Research and Innovation Program (Blue Growth: Unlocking the potential of Seas and Oceans) through the Project INMARE [634486]
- Ministry of Culture and Science of the German State of North Rhine-Westphalia [313/323-400-00213]
- Ministry of Science, Innovation and Universities [BIO2017-85522-R]
- ERDF
- Deutsche Forschungsgemeinschaft (DFG) [417919780]
- Agencia Estatal de Investigacion (AEI)
Biodegradation of synthetic polymers, in particular polyethylene terephthalate (PET), is of great importance, since environmental pollution with PET and other plastics has become a severe global problem. Here, we report on the polyester degrading ability of a novel carboxylic ester hydrolase identified in the genome of the marine hydrocarbonoclastic bacterium Pseudomonas aestusnigri VGXO14(T). The enzyme, designated PE-H, belongs to the type IIa family of PET hydrolytic enzymes as indicated by amino acid sequence homology. It was produced in Escherichia coli, purified and its crystal structure was solved at 1.09 angstrom resolution representing the first structure of a type IIa PET hydrolytic enzyme. The structure shows a typical alpha/beta-hydrolase fold and high structural homology to known polyester hydrolases. PET hydrolysis was detected at 30 degrees C with amorphous PET film (PETa), but not with PET film from a commercial PET bottle (PETb). A rational mutagenesis study to improve the PET degrading potential of PE-H yielded variant PE-H (Y250S) which showed improved activity, ultimately also allowing the hydrolysis of PETb. The crystal structure of this variant solved at 1.35 angstrom resolution allowed to rationalize the improvement of enzymatic activity. A PET oligomer binding model was proposed by molecular docking computations. Our results indicate a significant potential of the marine bacterium P. aestusnigri for PET degradation.
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