PpEst is a novel PBAT degrading polyesterase identified by proteomic screening of Pseudomonas pseudoalcaligenes

Abstract

A novel esterase, PpEst, that hydrolyses the co-aromatic-aliphatic polyester poly(1,4-butylene adipate-co-terephthalate) (PBAT) was identified by proteomic screening of the Pseudomonas pseudoalcaligenes secretome. PpEst was induced by the presence of PBAT in the growth media and had predicted arylesterase (EC 3.1.1.2) activity. PpEst showed polyesterase activity on both whole and milled PBAT film releasing terephthalic acid and 4-(4-hydroxybutoxycarbonyl)benzoic acid while end product inhibition by 4-(4-hydroxybutoxycarbonyl)benzoic acid was observed. Modelling of an aromatic polyester mimicking oligomer into the PpEst active site indicated that the binding pocket could be big enough to accommodate large polymers. This is the first report of a PBAT degrading enzyme being identified by proteomic screening and shows that this approach can contribute to the discovery of new polymer hydrolysing enzymes. Moreover, these results indicate that arylesterases could be an interesting enzyme class for identifications of polyesterases.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-016-7992-8) contains supplementary material, which is available to authorized users.

Shotgun proteomics reveals putative polyesterases in the secretome of the rock-inhabiting fungus Knufia chersonesos

“…The monomer Ta represented the most abundant compound detected at all experimental conditions and was proven not to be metabolised by the organism. This suggests the involvement of the fungus extracellular enzymes in the cleavage of aromatic ester bonds present in PBAT chains 17,45 , which are reportedly less prone to hydrolysis than the ester bond with aliphatic monomer (adipic acid) as also described for P. pseudoalcaligenes polyesterase PpEst 15 . Previous studies on single enzyme hydrolysis process of milled PBAT reported that BTaB and BTa were more abundant than the monomer, as well as that an increase of Ta concentration is given by higher pH than that selected in the current study 7 .…”

“…Microbial activities against PBAT have been shown in bacteria and to a lesser extent in fungi from soil [13][14][15][16][17] , by esterolytic hydrolases 7,15,18,19 . Typically, the investigation of PBAT degrading enzymes involves functional screenings and protein isolation through chromatographic purification 14,16,20,21 .…”

“…Alternatively, in silico screenings of OMICS databases are performed prior to heterologous expression of the target enzymes 12 . With only a few exceptions 15 , the majority of the studies however lack a comprehensive investigation at the proteome level of the possible effects of the polymer on induction of enzyme production by microorganisms. Degradation of polyesters www.nature.com/scientificreports www.nature.com/scientificreports/ is largely influenced by microbial activity in the environment and more directly by the asset of secreted hydrolytic enzymes.…”

The “Plastisphere” of Biodegradable Plastics Is Characterized by Specific Microbial Taxa of Alpine and Arctic Soils

“…Members of these orders were also found among the isolated strains obtained from the incubated plastics. Especially Pseudomonas is well known for its broad metabolic capabilities, and various strains of the genus are associated with the degradation of PLA, PBAT, PE, and many other plastics (Wallace et al, 2017;Wilkes and Aristilde, 2017;Bubpachat et al, 2018). Burkholderiales possess a broad range of enzymes able to degrade polyaromatic hydrocarbons (PAHs), which include important soil pollutants (Pérez-Pantoja et al, 2012).…”

Agar plate‐based screening methods for the identification of polyester hydrolysis by Pseudomonas species

“…By using a hidden Markov Model‐based search strategy to screen sequence data sets, Danso and co‐authors showed that a surprisingly large variety of potential polyesterases is still to be discovered, in particular in bacteria which are currently not considered as a prime source for cutinases (Danso et al ., ). Pseudomonas species may constitute an example; in the context of polymer hydrolysis, they appeared as a source for enzymes hydrolyzing polyurethane (Wilkes and Aristilde, ) for a long time, but some very recent reports by the Guebitz group indicated also polyesterase activity in Pseudomonads (Haernvall et al ., ,; Wallace et al ., ). The here reported confirmation of polyesterase activity of bacteria from the P. pertucinogena lineage, that was already suggested by sequence homology searches (Bollinger et al ., ), underlines the biotechnological potential of this group of bacteria.…”