Identification and characterization of an acyl-CoA dehydrogenase from Pseudomonas putida KT2440 that shows preference towards medium to long chain length fatty acids
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Guzik, MaciejNarančić, Tanja
Ilić-Tomić, Tatjana
Vojnović, Sandra
Kenny, Shane T.
Casey, William T.
Duane, Gearoid F.
Casey, Eoin
Woods, Trevor
Babu, Ramesh P.
Nikodinović-Runić, Jasmina
O'Connor, Kevin E.
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Diverse and elaborate pathways for nutrient utilization, as well as mechanisms to combat unfavourable nutrient conditions make Pseudomonas putida KT2440 a versatile micro-organism able to occupy a range of ecological niches. The fatty acid degradation pathway of P. putida is complex and correlated with biopolymer medium chain length polyhydroxyalkanoate (mcl-PHA) biosynthesis. Little is known about the second step of fatty acid degradation (beta-oxidation) in this strain. In silico analysis of its genome sequence revealed 21 putative acyl-CoA dehydrogenases (ACADs), four of which were functionally characterized through mutagenesis studies. Four mutants with insertionally inactivated ACADs (PP_1893, PP_2039, PP_2048 and PP_2437) grew and accumulated mcl-PHA on a range of fatty acids as the sole source of carbon and energy. Their ability to grow and accumulate biopolymer was differentially negatively affected on various fatty acids, in comparison to the wild-type strain. Inactive PP_2437... exhibited a pattern of reduced growth and PHA accumulation when fatty acids with lengths of 10 to 14 carbon chains were used as substrates. Recombinant expression and biochemical characterization of the purified protein allowed functional annotation in P. putida KT2440 as an ACAD showing clear preference for dodecanoyl-CoA ester as a substrate and optimum activity at 30 degrees C and pH 6.5-7.
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Microbiology, SGM / Society for General Microbiology, 2014, 160, 1760-1771Publisher:
- Soc General Microbiology, Reading
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- Free full text: https://dx.doi.org/10.1099/mic.0.078758-0
DOI: 10.1099/mic.0.078758-0
ISSN: 1350-0872
PubMed: 24794972
WoS: 000341678900019
Scopus: 2-s2.0-84905270409
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Inovacioni centar / Innovation CentreTY - JOUR AU - Guzik, Maciej AU - Narančić, Tanja AU - Ilić-Tomić, Tatjana AU - Vojnović, Sandra AU - Kenny, Shane T. AU - Casey, William T. AU - Duane, Gearoid F. AU - Casey, Eoin AU - Woods, Trevor AU - Babu, Ramesh P. AU - Nikodinović-Runić, Jasmina AU - O'Connor, Kevin E. PY - 2014 UR - https://cherry.chem.bg.ac.rs/handle/123456789/1846 AB - Diverse and elaborate pathways for nutrient utilization, as well as mechanisms to combat unfavourable nutrient conditions make Pseudomonas putida KT2440 a versatile micro-organism able to occupy a range of ecological niches. The fatty acid degradation pathway of P. putida is complex and correlated with biopolymer medium chain length polyhydroxyalkanoate (mcl-PHA) biosynthesis. Little is known about the second step of fatty acid degradation (beta-oxidation) in this strain. In silico analysis of its genome sequence revealed 21 putative acyl-CoA dehydrogenases (ACADs), four of which were functionally characterized through mutagenesis studies. Four mutants with insertionally inactivated ACADs (PP_1893, PP_2039, PP_2048 and PP_2437) grew and accumulated mcl-PHA on a range of fatty acids as the sole source of carbon and energy. Their ability to grow and accumulate biopolymer was differentially negatively affected on various fatty acids, in comparison to the wild-type strain. Inactive PP_2437 exhibited a pattern of reduced growth and PHA accumulation when fatty acids with lengths of 10 to 14 carbon chains were used as substrates. Recombinant expression and biochemical characterization of the purified protein allowed functional annotation in P. putida KT2440 as an ACAD showing clear preference for dodecanoyl-CoA ester as a substrate and optimum activity at 30 degrees C and pH 6.5-7. PB - Soc General Microbiology, Reading T2 - Microbiology, SGM / Society for General Microbiology T1 - Identification and characterization of an acyl-CoA dehydrogenase from Pseudomonas putida KT2440 that shows preference towards medium to long chain length fatty acids VL - 160 SP - 1760 EP - 1771 DO - 10.1099/mic.0.078758-0 ER -
@article{ author = "Guzik, Maciej and Narančić, Tanja and Ilić-Tomić, Tatjana and Vojnović, Sandra and Kenny, Shane T. and Casey, William T. and Duane, Gearoid F. and Casey, Eoin and Woods, Trevor and Babu, Ramesh P. and Nikodinović-Runić, Jasmina and O'Connor, Kevin E.", year = "2014", abstract = "Diverse and elaborate pathways for nutrient utilization, as well as mechanisms to combat unfavourable nutrient conditions make Pseudomonas putida KT2440 a versatile micro-organism able to occupy a range of ecological niches. The fatty acid degradation pathway of P. putida is complex and correlated with biopolymer medium chain length polyhydroxyalkanoate (mcl-PHA) biosynthesis. Little is known about the second step of fatty acid degradation (beta-oxidation) in this strain. In silico analysis of its genome sequence revealed 21 putative acyl-CoA dehydrogenases (ACADs), four of which were functionally characterized through mutagenesis studies. Four mutants with insertionally inactivated ACADs (PP_1893, PP_2039, PP_2048 and PP_2437) grew and accumulated mcl-PHA on a range of fatty acids as the sole source of carbon and energy. Their ability to grow and accumulate biopolymer was differentially negatively affected on various fatty acids, in comparison to the wild-type strain. Inactive PP_2437 exhibited a pattern of reduced growth and PHA accumulation when fatty acids with lengths of 10 to 14 carbon chains were used as substrates. Recombinant expression and biochemical characterization of the purified protein allowed functional annotation in P. putida KT2440 as an ACAD showing clear preference for dodecanoyl-CoA ester as a substrate and optimum activity at 30 degrees C and pH 6.5-7.", publisher = "Soc General Microbiology, Reading", journal = "Microbiology, SGM / Society for General Microbiology", title = "Identification and characterization of an acyl-CoA dehydrogenase from Pseudomonas putida KT2440 that shows preference towards medium to long chain length fatty acids", volume = "160", pages = "1760-1771", doi = "10.1099/mic.0.078758-0" }
Guzik, M., Narančić, T., Ilić-Tomić, T., Vojnović, S., Kenny, S. T., Casey, W. T., Duane, G. F., Casey, E., Woods, T., Babu, R. P., Nikodinović-Runić, J.,& O'Connor, K. E.. (2014). Identification and characterization of an acyl-CoA dehydrogenase from Pseudomonas putida KT2440 that shows preference towards medium to long chain length fatty acids. in Microbiology, SGM / Society for General Microbiology Soc General Microbiology, Reading., 160, 1760-1771. https://doi.org/10.1099/mic.0.078758-0
Guzik M, Narančić T, Ilić-Tomić T, Vojnović S, Kenny ST, Casey WT, Duane GF, Casey E, Woods T, Babu RP, Nikodinović-Runić J, O'Connor KE. Identification and characterization of an acyl-CoA dehydrogenase from Pseudomonas putida KT2440 that shows preference towards medium to long chain length fatty acids. in Microbiology, SGM / Society for General Microbiology. 2014;160:1760-1771. doi:10.1099/mic.0.078758-0 .
Guzik, Maciej, Narančić, Tanja, Ilić-Tomić, Tatjana, Vojnović, Sandra, Kenny, Shane T., Casey, William T., Duane, Gearoid F., Casey, Eoin, Woods, Trevor, Babu, Ramesh P., Nikodinović-Runić, Jasmina, O'Connor, Kevin E., "Identification and characterization of an acyl-CoA dehydrogenase from Pseudomonas putida KT2440 that shows preference towards medium to long chain length fatty acids" in Microbiology, SGM / Society for General Microbiology, 160 (2014):1760-1771, https://doi.org/10.1099/mic.0.078758-0 . .