Catalysis

Pseudomonas

Bacteria

Biology

Identification (biology)

Materials science

Chromatography

Biodegradation

Cutin

Polyhydroxyalkanoates

Biochemistry

Botany

Polyester

Polymer

Genetics

Combinatorial chemistry

Agar

Organic chemistry

Biocatalysis

Polycaprolactone

Microbiology

Hydrolysis

Composite material

Species identification

Chemistry

Ionic liquid

Agar plate

Polyethylene terephthalate

Microbial Biotechnology

Hydrolases acting on polyesters like cutin, polycaprolactone or polyethylene terephthalate (PET) are of interest for several biotechnological applications like waste treatment, biocatalysis and sustainable polymer modifications. Recent studies suggest that a large variety of such enzymes are still to be identified and explored in a variety of microorganisms, including bacteria of the genus Pseudomonas. For activity-based screening, methods have been established using agar plates which contain nanoparticles of polycaprolactone or PET prepared by solvent precipitation and evaporation. In this protocol article, we describe a straightforward agar plate-based method using emulsifiable artificial polyesters as substrates, namely Impranil® DLN and liquid polycaprolactone diol (PLD). Thereby, the currently quite narrow set of screening substrates is expanded. We also suggest optional pre-screening with short-chain and middle-chain-length triglycerides as substrates to identify enzymes with lipolytic activity to be further tested for polyesterase activity. We applied these assays to experimentally demonstrate polyesterase activity in bacteria from the P. pertucinogena lineage originating from contaminated soils and diverse marine habitats.

Agar plate‐based screening methods for the identification of polyester hydrolysis by <i>Pseudomonas</i> species

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