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The effect of polyphosphate kinase gene deletion on polyhydroxyalkanoate accumulation and carbon metabolism in Pseudomonas putida KT2440

Authorized Users Only
2013
Authors
Casey, William T.
Nikodinović-Runić, Jasmina
Fonseca Garcia, Pilar
Guzik, Maciej
McGrath, John W.
Quinn, John P.
Cagney, Gerard
Auxiliadora Prieto, Maria
O'Connor, Kevin E.
Article (Published version)
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Abstract
The primary enzyme involved in polyphosphate (polyP) synthesis, polyP kinase (ppk), has been deleted in Pseudomonas putidaKT2440. This has resulted in a threefold to sixfold reduction in polyhydroxyalkanoate (PHA) accumulation compared with the wild type under conditions of nitrogen limitation, with either temperature or oxidative (H2O2) stress, when grown on glucose. The accumulation of PHA by ppk mutant was the same as the wild type under nitrogen-limiting growth conditions. There was no difference in polyP levels between wild-type and ppk strains under all growth conditions tested. In the ppk mutant proteome, polyP kinase (PPK) was undetectable, but up-regulation of the polyp-associated proteins polyP adenosine triphosphate (ATP)/nicotinamide adenine dinucleotide (NAD) kinase (PpnK), a putative polyP adenosine monophosphate (AMP) phosphotransferase (PP_1752), and exopolyphosphatase was observed. ppk strain exhibited significantly retarded growth with glycerol as carbon and energy so...urce (42h of lag period compared with 24h in wild-type strain) but similar growth to the wild-type strain with glucose. Analysis of gene transcription revealed downregulation of glycerol kinase and the glycerol facilitator respectively. Glycerol kinase protein expression was also downregulated in the ppk mutant. The deletion of ppk did not affect motility but reduced biofilm formation. Thus, the knockout of the ppk gene has resulted in a number of phenotypic changes to the mutant without affecting polyP accumulation.

Source:
Environmental Microbiology Reports, 2013, 5, 5, 740-746
Publisher:
  • Wiley-Blackwell, Hoboken
Note:
  • Peer-reviewed manuscript: http://cherry.chem.bg.ac.rs/handle/123456789/3553
  • Supplementary material: http://cherry.chem.bg.ac.rs/handle/123456789/3554

DOI: 10.1111/1758-2229.12076

ISSN: 1758-2229

PubMed: 24115625

WoS: 000325142700014

Scopus: 2-s2.0-84884973014
[ Google Scholar ]
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URI
http://cherry.chem.bg.ac.rs/handle/123456789/1408
Collections
  • Publikacije
Institution
Inovacioni centar
TY  - JOUR
AU  - Casey, William T.
AU  - Nikodinović-Runić, Jasmina
AU  - Fonseca Garcia, Pilar
AU  - Guzik, Maciej
AU  - McGrath, John W.
AU  - Quinn, John P.
AU  - Cagney, Gerard
AU  - Auxiliadora Prieto, Maria
AU  - O'Connor, Kevin E.
PY  - 2013
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/1408
AB  - The primary enzyme involved in polyphosphate (polyP) synthesis, polyP kinase (ppk), has been deleted in Pseudomonas putidaKT2440. This has resulted in a threefold to sixfold reduction in polyhydroxyalkanoate (PHA) accumulation compared with the wild type under conditions of nitrogen limitation, with either temperature or oxidative (H2O2) stress, when grown on glucose. The accumulation of PHA by ppk mutant was the same as the wild type under nitrogen-limiting growth conditions. There was no difference in polyP levels between wild-type and ppk strains under all growth conditions tested. In the ppk mutant proteome, polyP kinase (PPK) was undetectable, but up-regulation of the polyp-associated proteins polyP adenosine triphosphate (ATP)/nicotinamide adenine dinucleotide (NAD) kinase (PpnK), a putative polyP adenosine monophosphate (AMP) phosphotransferase (PP_1752), and exopolyphosphatase was observed. ppk strain exhibited significantly retarded growth with glycerol as carbon and energy source (42h of lag period compared with 24h in wild-type strain) but similar growth to the wild-type strain with glucose. Analysis of gene transcription revealed downregulation of glycerol kinase and the glycerol facilitator respectively. Glycerol kinase protein expression was also downregulated in the ppk mutant. The deletion of ppk did not affect motility but reduced biofilm formation. Thus, the knockout of the ppk gene has resulted in a number of phenotypic changes to the mutant without affecting polyP accumulation.
PB  - Wiley-Blackwell, Hoboken
T2  - Environmental Microbiology Reports
T1  - The effect of polyphosphate kinase gene deletion on polyhydroxyalkanoate accumulation and carbon metabolism in Pseudomonas putida KT2440
VL  - 5
IS  - 5
SP  - 740
EP  - 746
DO  - 10.1111/1758-2229.12076
ER  - 
@article{
author = "Casey, William T. and Nikodinović-Runić, Jasmina and Fonseca Garcia, Pilar and Guzik, Maciej and McGrath, John W. and Quinn, John P. and Cagney, Gerard and Auxiliadora Prieto, Maria and O'Connor, Kevin E.",
year = "2013",
url = "http://cherry.chem.bg.ac.rs/handle/123456789/1408",
abstract = "The primary enzyme involved in polyphosphate (polyP) synthesis, polyP kinase (ppk), has been deleted in Pseudomonas putidaKT2440. This has resulted in a threefold to sixfold reduction in polyhydroxyalkanoate (PHA) accumulation compared with the wild type under conditions of nitrogen limitation, with either temperature or oxidative (H2O2) stress, when grown on glucose. The accumulation of PHA by ppk mutant was the same as the wild type under nitrogen-limiting growth conditions. There was no difference in polyP levels between wild-type and ppk strains under all growth conditions tested. In the ppk mutant proteome, polyP kinase (PPK) was undetectable, but up-regulation of the polyp-associated proteins polyP adenosine triphosphate (ATP)/nicotinamide adenine dinucleotide (NAD) kinase (PpnK), a putative polyP adenosine monophosphate (AMP) phosphotransferase (PP_1752), and exopolyphosphatase was observed. ppk strain exhibited significantly retarded growth with glycerol as carbon and energy source (42h of lag period compared with 24h in wild-type strain) but similar growth to the wild-type strain with glucose. Analysis of gene transcription revealed downregulation of glycerol kinase and the glycerol facilitator respectively. Glycerol kinase protein expression was also downregulated in the ppk mutant. The deletion of ppk did not affect motility but reduced biofilm formation. Thus, the knockout of the ppk gene has resulted in a number of phenotypic changes to the mutant without affecting polyP accumulation.",
publisher = "Wiley-Blackwell, Hoboken",
journal = "Environmental Microbiology Reports",
title = "The effect of polyphosphate kinase gene deletion on polyhydroxyalkanoate accumulation and carbon metabolism in Pseudomonas putida KT2440",
volume = "5",
number = "5",
pages = "740-746",
doi = "10.1111/1758-2229.12076"
}
Casey WT, Nikodinović-Runić J, Fonseca Garcia P, Guzik M, McGrath JW, Quinn JP, Cagney G, Auxiliadora Prieto M, O'Connor KE. The effect of polyphosphate kinase gene deletion on polyhydroxyalkanoate accumulation and carbon metabolism in Pseudomonas putida KT2440. Environmental Microbiology Reports. 2013;5(5):740-746
Casey, W. T., Nikodinović-Runić, J., Fonseca Garcia, P., Guzik, M., McGrath, J. W., Quinn, J. P., Cagney, G., Auxiliadora Prieto, M.,& O'Connor, K. E. (2013). The effect of polyphosphate kinase gene deletion on polyhydroxyalkanoate accumulation and carbon metabolism in Pseudomonas putida KT2440.
Environmental Microbiology ReportsWiley-Blackwell, Hoboken., 5(5), 740-746.
https://doi.org/10.1111/1758-2229.12076
Casey William T., Nikodinović-Runić Jasmina, Fonseca Garcia Pilar, Guzik Maciej, McGrath John W., Quinn John P., Cagney Gerard, Auxiliadora Prieto Maria, O'Connor Kevin E., "The effect of polyphosphate kinase gene deletion on polyhydroxyalkanoate accumulation and carbon metabolism in Pseudomonas putida KT2440" 5, no. 5 (2013):740-746,
https://doi.org/10.1111/1758-2229.12076 .

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