An analysis of the changes in soluble hydrogenase and global gene expression in Cupriavidus necator (Ralstonia eutropha) H16 grown in heterotrophic diauxic batch culture

We have cloned, overexpressed, purified, and characterized a 2-ketogluconate kinase (2-dehydrogluconokinase, EC 2.7.1.13) from Cupriavidus necator (Ralstonia eutropha) H16. Exploration of its substrate specificity revealed that three ketoacids (2-keto-3-deoxy-d-gluconate, 2-keto-d-gulonate, and 2-keto-3-deoxy-d-gulonate) with structures close to the natural substrate (2-keto-d-gluconate) were successfully phosphorylated at an efficiency lower than or comparable to 2-ketogluconate, as depicted by the measured kinetic constant values. Eleven aldo and keto monosaccharides of different chain lengths and stereochemistries were also assayed but not found to be substrates. 2-ketogluconate-6-phosphate was synthesized at a preparative scale and was fully characterized for the first time.

Download Full-text

Versatile and stable vectors for efficient gene expression in Ralstonia eutropha H16

New Biotechnology ◽

10.1016/j.nbt.2014.05.1776 ◽

2014 ◽

Vol 31 ◽

pp. S72

Author(s):

Steffen Gruber ◽

Jeremias Hagen ◽

Helmut Schwab ◽

Petra Koefinger

Keyword(s):

Gene Expression ◽

Ralstonia Eutropha ◽

Efficient Gene ◽

Ralstonia Eutropha H16 ◽

Stable Vectors

Download Full-text

Versatile and stable vectors for efficient gene expression in Ralstonia eutropha H16

Journal of Biotechnology ◽

10.1016/j.jbiotec.2014.06.030 ◽

2014 ◽

Vol 186 ◽

pp. 74-82 ◽

Cited By ~ 23

Author(s):

Steffen Gruber ◽

Jeremias Hagen ◽

Helmut Schwab ◽

Petra Koefinger

Keyword(s):

Gene Expression ◽

Ralstonia Eutropha ◽

Efficient Gene ◽

Ralstonia Eutropha H16 ◽

Stable Vectors

Download Full-text

Reprint of “Versatile and stable vectors for efficient gene expression in Ralstonia eutropha H16”

Journal of Biotechnology ◽

10.1016/j.jbiotec.2014.09.023 ◽

2014 ◽

Vol 192 ◽

pp. 410-418 ◽

Cited By ~ 3

Author(s):

Steffen Gruber ◽

Jeremias Hagen ◽

Helmut Schwab ◽

Petra Koefinger

Keyword(s):

Gene Expression ◽

Ralstonia Eutropha ◽

Efficient Gene ◽

Ralstonia Eutropha H16 ◽

Stable Vectors

Download Full-text

Elucidation of β-Oxidation Pathways in Ralstonia eutropha H16 by Examination of Global Gene Expression

Journal of Bacteriology ◽

10.1128/jb.00493-10 ◽

2010 ◽

Vol 192 (20) ◽

pp. 5454-5464 ◽

Cited By ~ 74

Author(s):

Christopher J. Brigham ◽

Charles F. Budde ◽

Jason W. Holder ◽

Qiandong Zeng ◽

Alison E. Mahan ◽

...

Keyword(s):

Gene Expression ◽

Fatty Acid ◽

Deletion Mutant ◽

Ralstonia Eutropha ◽

Isocitrate Lyase ◽

Palm Kernel ◽

Global Gene Expression ◽

Malate Synthase ◽

Growth Defect ◽

Plant Oils

ABSTRACT Ralstonia eutropha H16 is capable of growth and polyhydroxyalkanoate production on plant oils and fatty acids. However, little is known about the triacylglycerol and fatty acid degradation pathways of this bacterium. We compare whole-cell gene expression levels of R. eutropha H16 during growth and polyhydroxyalkanoate production on trioleate and fructose. Trioleate is a triacylglycerol that serves as a model for plant oils. Among the genes of note, two potential fatty acid β-oxidation operons and two putative lipase genes were shown to be upregulated in trioleate cultures. The genes of the glyoxylate bypass also exhibit increased expression during growth on trioleate. We observed that single β-oxidation operon deletion mutants of R. eutropha could grow using palm oil or crude palm kernel oil as the sole carbon source, regardless of which operon was present in the genome, but a double mutant was unable to grow under these conditions. A lipase deletion mutant did not exhibit a growth defect in emulsified oil cultures but did exhibit a phenotype in cultures containing nonemulsified oil. Mutants of the glyoxylate shunt gene for isocitrate lyase were able to grow in the presence of oils, while a malate synthase (aceB) deletion mutant grew more slowly than wild type. Gene expression under polyhydroxyalkanoate storage conditions was also examined. Many findings of this analysis confirm results from previous studies by our group and others. This work represents the first examination of global gene expression involving triacylglycerol and fatty acid catabolism genes in R. eutropha.

Download Full-text

NO-dependent transcriptional activation of gene expression in Ralstonia eutropha H16

Biochemical Society Transactions ◽

10.1042/bst0340182 ◽

2006 ◽

Vol 34 (1) ◽

pp. 182-184 ◽

Cited By ~ 6

Author(s):

R. Cramm ◽

A. Büsch ◽

K. Strube

Keyword(s):

Gene Expression ◽

Transcriptional Activation ◽

Ralstonia Eutropha ◽

Promoter Activation ◽

Iron Protein ◽

Signal Perception ◽

Redox Active ◽

Negative Effect ◽

Ralstonia Eutropha H16 ◽

Haem Iron

The σ54-dependent transcriptional regulator NorR of Ralstonia eutropha H16 activates gene expression in response to nitric oxide (NO). The N-terminal domain of NorR is thought to be involved in signal perception. A C112S exchange within this domain abolished promoter activation by the mutated protein, indicating that Cys112 is essential for the signalling mechanism of NorR. The DNA region recognized by NorR contains three copies of a conserved element termed the NorR-box. Alteration of bases within any of the NorR-boxes resulted in a significant decrease in promoter activation. Therefore all three boxes have to be recognized by NorR to activate its target promoter. NorR controls expression of an operon that encodes a redox-active non-haem-iron protein NorA and an NO reductase NorB. NorA exerts a negative effect on signal-dependent promoter activation by NorR. Optical spectroscopy of purified NorA indicates that the reduced protein can react with NO to form a ferrous nitrosyl adduct. Hence, NO binding by NorA opens up the possibility that NorA and NorR compete for NO in the cytoplasm.

Download Full-text

Transcriptional profiling of iPSC-derived neurons with reciprocal monogenic CNV in 3p26.3 region

Nauchno-prakticheskii zhurnal «Medicinskaia genetika» ◽

10.25557/2073-7998.2020.03.10-11 ◽

2020 ◽

pp. 10-11

Author(s):

М.Е. Лопаткина ◽

В.С. Фишман ◽

М.М. Гридина ◽

Н.А. Скрябин ◽

Т.В. Никитина ◽

...

Keyword(s):

Gene Expression ◽

Copy Number ◽

System Development ◽

Transcriptional Profiling ◽

Global Gene Expression ◽

Nervous System Development ◽

Number Variation ◽

The Central Nervous System ◽

Cntn6 Gene ◽

Copy Number Changes

Проведен анализ генной экспрессии в нейронах, дифференцированных из индуцированных плюрипотентных стволовых клеток пациентов с идиопатическими интеллектуальными нарушениями и реципрокными хромосомными мутациями в регионе 3p26.3, затрагивающими единственный ген CNTN6. Для нейронов с различным типом хромосомных аберраций была показана глобальная дисрегуляция генной экспрессии. В нейронах с вариациями числа копий гена CNTN6 была снижена экспрессия генов, продукты которых вовлечены в процессы развития центральной нервной системы. The gene expression analysis of iPSC-derived neurons, obtained from patients with idiopathic intellectual disability and reciprocal microdeletion and microduplication in 3p26.3 region affecting the single CNTN6 gene was performed. The global gene expression dysregulation was demonstrated for cells with CNTN6 copy number variation. Gene expression in neurons with CNTN6 copy number changes was downregulated for genes, whose products are involved in the central nervous system development.

Download Full-text