Medium chain length polyhyoxyalkanoates (mcl-PHA) model compounds for the discovery of novel PHA depolymerases
Аутори
Nenadović, MarijaPantelić, Brana
Lazić, Jelena
Maslak, Veselin
Nikodinović-Runić, Jasmina
Milovanović, Jelena
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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.
Извор:
10th Conference of Mikrobiokosmos, 2023Финансирање / пројекти:
- European Union's Horizon 2020 research and innovation program under grant agreement No. 870292
Напомена:
- 10th International Conference of MIKROBIOKOSMOS, Larissa from 30 Novewmber to 2 December 2023.
URI
https://afea.eventsair.com/10th-conference-of-mikrobiokosmos/abstract-bookhttps://imagine.imgge.bg.ac.rs/handle/123456789/2237
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - 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 .