TY  - CONF
AU  - Nenadović, Marija
AU  - Pantelić, Brana
AU  - Lazić, Jelena
AU  - Maslak, Veselin
AU  - Nikodinović-Runić, Jasmina
AU  - Milovanović, Jelena
PY  - 2023
UR  - https://afea.eventsair.com/10th-conference-of-mikrobiokosmos/abstract-book
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2237
AB  - PHAs are naturally made microbial polyesters that
have been commercialized as biodegradable
plastics. However, it has been shown that these
materials are not so easily biodegraded in natural
environments [1]. PHA depolymerases are key PHA
degrading enzymes and their identification and
characterization is of great interest and importance.
Currently, screening is done on polymeric
substrates using techniques such as clear zone
assays on agar or weight loss measurements.
Results obtained using these different methods
cannot be directly compared, since they depend
highly on the polymer used, PHA granules
preparation and assay conditions [2].
In order to design a more specific test for the
determination of PHA depolymerase activity, we
synthesized 3-hyoxyalkanoate monomers (3-HA
monomer) and 3-hyoxyalkanoic acid dimers (3-HA
dimer) and their respective p-nitrophenyl esters,
allowing for spectrophotometric determination of
their activity [3]. Compounds were characterized
using N and FTIR. Para-nitrophenyl labeled
substrates were then used in the enzymatic activity
assay with the benchmark polyhyoxyoctanoate
(PHO) depolymerase from Pseudomonas
fluorescens GK13 expressed in Escherichia coli
CodonPlus-RIPL hosts. This activity was compared
to recombinantly expressed leaf-branch compost
cutinase (LCC cutinase) and
polyethyleneterephtalate (PET) hyolyzing esterase
from Ideonella sakaiensis (IsPETase). Our initial
results revealed increased specificity of PHO
depolymerase towards newly synthetized
substrates, suggesting their suitability for specific
screens and isolation of new mcl-PHA
depolymerases, as well as in high throughput
screening assays designed for guiding their
directed evolution.
C3  - 10th Conference of Mikrobiokosmos
T1  - Medium chain length polyhyoxyalkanoates (mcl-PHA) model compounds for the discovery of novel PHA depolymerases
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2237
ER  - 

@conference{
author = "Nenadović, Marija and Pantelić, Brana and Lazić, Jelena and Maslak, Veselin and Nikodinović-Runić, Jasmina and Milovanović, Jelena",
year = "2023",
abstract = "PHAs are naturally made microbial polyesters that
have been commercialized as biodegradable
plastics. However, it has been shown that these
materials are not so easily biodegraded in natural
environments [1]. PHA depolymerases are key PHA
degrading enzymes and their identification and
characterization is of great interest and importance.
Currently, screening is done on polymeric
substrates using techniques such as clear zone
assays on agar or weight loss measurements.
Results obtained using these different methods
cannot be directly compared, since they depend
highly on the polymer used, PHA granules
preparation and assay conditions [2].
In order to design a more specific test for the
determination of PHA depolymerase activity, we
synthesized 3-hyoxyalkanoate monomers (3-HA
monomer) and 3-hyoxyalkanoic acid dimers (3-HA
dimer) and their respective p-nitrophenyl esters,
allowing for spectrophotometric determination of
their activity [3]. Compounds were characterized
using N and FTIR. Para-nitrophenyl labeled
substrates were then used in the enzymatic activity
assay with the benchmark polyhyoxyoctanoate
(PHO) depolymerase from Pseudomonas
fluorescens GK13 expressed in Escherichia coli
CodonPlus-RIPL hosts. This activity was compared
to recombinantly expressed leaf-branch compost
cutinase (LCC cutinase) and
polyethyleneterephtalate (PET) hyolyzing esterase
from Ideonella sakaiensis (IsPETase). Our initial
results revealed increased specificity of PHO
depolymerase towards newly synthetized
substrates, suggesting their suitability for specific
screens and isolation of new mcl-PHA
depolymerases, as well as in high throughput
screening assays designed for guiding their
directed evolution.",
journal = "10th Conference of Mikrobiokosmos",
title = "Medium chain length polyhyoxyalkanoates (mcl-PHA) model compounds for the discovery of novel PHA depolymerases",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2237"
}

Nenadović, M., Pantelić, B., Lazić, J., Maslak, V., Nikodinović-Runić, J.,& Milovanović, J.. (2023). Medium chain length polyhyoxyalkanoates (mcl-PHA) model compounds for the discovery of novel PHA depolymerases. in 10th Conference of Mikrobiokosmos.
https://hdl.handle.net/21.15107/rcub_imagine_2237

Nenadović M, Pantelić B, Lazić J, Maslak V, Nikodinović-Runić J, Milovanović J. Medium chain length polyhyoxyalkanoates (mcl-PHA) model compounds for the discovery of novel PHA depolymerases. in 10th Conference of Mikrobiokosmos. 2023;.
https://hdl.handle.net/21.15107/rcub_imagine_2237 .

Nenadović, Marija, Pantelić, Brana, Lazić, Jelena, Maslak, Veselin, Nikodinović-Runić, Jasmina, Milovanović, Jelena, "Medium chain length polyhyoxyalkanoates (mcl-PHA) model compounds for the discovery of novel PHA depolymerases" in 10th Conference of Mikrobiokosmos (2023),
https://hdl.handle.net/21.15107/rcub_imagine_2237 .