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

Autor: Molitor, Rebecka, Bollinger, Alexander, Kubicki, Sonja, Loeschcke, Anita, Jaeger, Karl‐Erich, Thies, Stephan
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Zdroj: Microbial Biotechnology
Microbial Biotechnology, Vol 13, Iss 1, Pp 274-284 (2020)
Microbial biotechnology 13(1), 274-284 (2020). doi:10.1111/1751-7915.13418
ISSN: 1751-7915
DOI: 10.1111/1751-7915.13418
Popis: Summary 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.
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. In this protocol article, we describe a straightforward agar plate based method using emulsifiable artificial polyesters as substrates. We applied these assays to experimentally demonstrate polyesterase activity in bacteria from the Pseudomonas pertucinogena lineage originating from contaminated soils and diverse marine habitats.
Databáze: OpenAIRE
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