Establishing highthroughput screening of engineered polyhydroxyoctanoate (PHO) depolymerase variants using novel PHO model compounds
Апстракт
Polyhydroxyoctanoate (PHO) is a biocompatible microbially produced polyester with elastomeric properties currently limited by its high production cost and poor
biodegradability in open environments. Enzymatic recycling of postconsumer PHO offers a biocyclable route to PHO utilization and poses a solution from both
ecological and economical aspects. Enzyme engineering enables industrial efficiency in polymer degradation. Therefore, we have created a mutant library for PHO
depolymerase from Pseudomonas fluorescens GK13 (PfPHOase) using errorprone PCR. To advance the speed of identification of PfPHOase variants with improved
PHO degradation rates highthroughput screening was performed spectroscopically with inhouse synthesized pnitrophenyl esters of 3hydroxyalkanoate monomer (3
HA monomer) and 3hydroxyalkanoic acid dimer (3HA dimer)1. Further, the degradation of PHO polymer for bestperforming PfPHOase variants was assessed on
commonly employed emulsified PHOagarose plate...s. The obtained results indicate the positive correlation between the degradation of novel PHO model compounds
with degradation of PHO polymer. Novel model compounds have been successfully employed for the identification of beneficial mutations for the advancement of
enzymatic PHO degradation.
Извор:
48th FEBS Congress, 2024, P-20-005Издавач:
- Federation of European Biochemical Societies
Финансирање / пројекти:
- European Union's Horizon 2020 research and inovation programme under grant agreement No 870292.
Колекције
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - CONF AU - Nenadović, Marija AU - Milovanović, Jelena AU - Maslak, Veselin AU - Nikodinović-Runić, Jasmina PY - 2024 UR - https://2024.febscongress.org/ UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2423 AB - Polyhydroxyoctanoate (PHO) is a biocompatible microbially produced polyester with elastomeric properties currently limited by its high production cost and poor biodegradability in open environments. Enzymatic recycling of postconsumer PHO offers a biocyclable route to PHO utilization and poses a solution from both ecological and economical aspects. Enzyme engineering enables industrial efficiency in polymer degradation. Therefore, we have created a mutant library for PHO depolymerase from Pseudomonas fluorescens GK13 (PfPHOase) using errorprone PCR. To advance the speed of identification of PfPHOase variants with improved PHO degradation rates highthroughput screening was performed spectroscopically with inhouse synthesized pnitrophenyl esters of 3hydroxyalkanoate monomer (3 HA monomer) and 3hydroxyalkanoic acid dimer (3HA dimer)1. Further, the degradation of PHO polymer for bestperforming PfPHOase variants was assessed on commonly employed emulsified PHOagarose plates. The obtained results indicate the positive correlation between the degradation of novel PHO model compounds with degradation of PHO polymer. Novel model compounds have been successfully employed for the identification of beneficial mutations for the advancement of enzymatic PHO degradation. PB - Federation of European Biochemical Societies C3 - 48th FEBS Congress T1 - Establishing highthroughput screening of engineered polyhydroxyoctanoate (PHO) depolymerase variants using novel PHO model compounds IS - P-20-005 UR - https://hdl.handle.net/21.15107/rcub_imagine_2423 ER -
@conference{ author = "Nenadović, Marija and Milovanović, Jelena and Maslak, Veselin and Nikodinović-Runić, Jasmina", year = "2024", abstract = "Polyhydroxyoctanoate (PHO) is a biocompatible microbially produced polyester with elastomeric properties currently limited by its high production cost and poor biodegradability in open environments. Enzymatic recycling of postconsumer PHO offers a biocyclable route to PHO utilization and poses a solution from both ecological and economical aspects. Enzyme engineering enables industrial efficiency in polymer degradation. Therefore, we have created a mutant library for PHO depolymerase from Pseudomonas fluorescens GK13 (PfPHOase) using errorprone PCR. To advance the speed of identification of PfPHOase variants with improved PHO degradation rates highthroughput screening was performed spectroscopically with inhouse synthesized pnitrophenyl esters of 3hydroxyalkanoate monomer (3 HA monomer) and 3hydroxyalkanoic acid dimer (3HA dimer)1. Further, the degradation of PHO polymer for bestperforming PfPHOase variants was assessed on commonly employed emulsified PHOagarose plates. The obtained results indicate the positive correlation between the degradation of novel PHO model compounds with degradation of PHO polymer. Novel model compounds have been successfully employed for the identification of beneficial mutations for the advancement of enzymatic PHO degradation.", publisher = "Federation of European Biochemical Societies", journal = "48th FEBS Congress", title = "Establishing highthroughput screening of engineered polyhydroxyoctanoate (PHO) depolymerase variants using novel PHO model compounds", number = "P-20-005", url = "https://hdl.handle.net/21.15107/rcub_imagine_2423" }
Nenadović, M., Milovanović, J., Maslak, V.,& Nikodinović-Runić, J.. (2024). Establishing highthroughput screening of engineered polyhydroxyoctanoate (PHO) depolymerase variants using novel PHO model compounds. in 48th FEBS Congress Federation of European Biochemical Societies.(P-20-005). https://hdl.handle.net/21.15107/rcub_imagine_2423
Nenadović M, Milovanović J, Maslak V, Nikodinović-Runić J. Establishing highthroughput screening of engineered polyhydroxyoctanoate (PHO) depolymerase variants using novel PHO model compounds. in 48th FEBS Congress. 2024;(P-20-005). https://hdl.handle.net/21.15107/rcub_imagine_2423 .
Nenadović, Marija, Milovanović, Jelena, Maslak, Veselin, Nikodinović-Runić, Jasmina, "Establishing highthroughput screening of engineered polyhydroxyoctanoate (PHO) depolymerase variants using novel PHO model compounds" in 48th FEBS Congress, no. P-20-005 (2024), https://hdl.handle.net/21.15107/rcub_imagine_2423 .