PUL ID

PUL0396

PubMed

21478317, Appl Environ Microbiol. 2011 Jun;77(11):3582-90. doi: 10.1128/AEM.00218-11. Epub 2011 Apr 8.

Characterization method

gene deletion mutant and growth assay

Genomic accession number

AM260479.1

Nucelotide position range

327968-338299

Substrate

N-acetylglucosamine

Loci

H16_A0310-H16_A0316

Species

Cupriavidus necator/106590

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

h16_A0310 H16_A0310 CAJ91461.1 0 - 723 (-) AM260479.1:327968-328691 -
nagF H16_A0311 CAJ91462.1 1115 - 3668 (+) AM260479.1:329083-331636 2.7.1.69
nagE H16_A0312 CAJ91463.1 3725 - 5435 (+) AM260479.1:331693-333403 2.7.1.69
nagC H16_A0313 CAJ91464.1 5551 - 6616 (+) AM260479.1:333519-334584 -
nagA H16_A0314 CAJ91465.1 6658 - 7762 (+) AM260479.1:334626-335730 3.5.1.25
nagB H16_A0315 CAJ91466.1 7754 - 8786 (+) AM260479.1:335722-336754 3.5.99.6
zwf1 H16_A0316 CAJ91467.1 8844 - 10332 (+) AM260479.1:336812-338300 1.1.1.49

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

h16_A0310 1 - 723 (-) TF: DBD-Pfam|GntR,DBD-SUPERFAMILY|0037767 No
nagF 1116 - 3668 (+) TC: gnl|TC-DB|Q9HXN5|4.A.1.1.15 Yes
nagE 3726 - 5435 (+) TC: gnl|TC-DB|Q9HXN4|4.A.1.1.15 Yes
nagC 5552 - 6616 (+) TC: gnl|TC-DB|C0LZS0|1.B.1.4.2 Yes
nagA 6659 - 7762 (+) CAZyme: CE9 Yes
nagB 7755 - 8786 (+) STP: STP|SIS,STP|SIS No
zwf1 8845 - 10332 (+) CDS No

PUL ID

PUL0396

PubMed

21478317, Appl Environ Microbiol. 2011 Jun;77(11):3582-90. doi: 10.1128/AEM.00218-11. Epub 2011 Apr 8.

Title

Effects of homologous phosphoenolpyruvate-carbohydrate phosphotransferase system proteins on carbohydrate uptake and poly(3-Hydroxybutyrate) accumulation in Ralstonia eutropha H16.

Author

Kaddor C, Steinbuchel A

Abstract

Seven gene loci encoding putative proteins of the phosphoenolpyruvate-carbohydrate phosphotransferase system (PEP-PTS) were identified in the genome of Ralstonia eutropha H16 by in silico analysis. Except the N-acetylglucosamine-specific PEP-PTS, an additional complete PEP-PTS is lacking in strain H16. Based on these findings, we generated single and multiple deletion mutants defective mainly in the PEP-PTS genes to investigate their influence on carbon source utilization, growth behavior, and poly(3-hydroxybutyrate) (PHB) accumulation. As supposed, the H16 DeltafrcACB and H16 DeltanagFEC mutants exhibited no growth when cultivated on fructose and N-acetylglucosamine, respectively. Furthermore, a transposon mutant with a ptsM-ptsH insertion site did not grow on both carbon sources. The observed phenotype was not complemented, suggesting that it results from an interaction of genes or a polar effect caused by the Tn5::mob insertion. ptsM, ptsH, and ptsI single, double, and triple mutants stored much less PHB than the wild type (about 10 to 39% [wt/wt] of cell dry weight) and caused reduced PHB production in mutants lacking the H16_A2203, H16_A0384, frcACB, or nagFEC genes. In contrast, mutant H16 DeltaH16_A0384 accumulated 11.5% (wt/wt) more PHB than the wild type when grown on gluconate and suppressed partially the negative effect of the ptsMHI deletion on PHB synthesis. Based on our experimental data, we discussed whether the PEP-PTS homologous proteins in R. eutropha H16 are exclusively involved in the complex sugar transport system or whether they are also involved in cellular regulatory functions of carbon and PHB metabolism.