TY  - JOUR
AU  - Molitor, Rebecka
AU  - Bollinger, Alexander
AU  - Kubicki, Sonja
AU  - Loeschcke, Anita
AU  - Jaeger, Karl‐Erich
AU  - Thies, Stephan
TI  - Agar plate‐based screening methods for the identification of polyester hydrolysis by Pseudomonas species
JO  - Microbial biotechnology
VL  - 13
IS  - 1
SN  - 1751-7915
CY  - Oxford
PB  - Wiley-Blackwell
M1  - FZJ-2019-05372
SP  - 274-284
PY  - 2020
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:31016871
UR  - <Go to ISI:>//WOS:000596580200024
DO  - DOI:10.1111/1751-7915.13418
UR  - https://juser.fz-juelich.de/record/866201
ER  -