scholarly journals Effect of lysine succinylation on the regulation of 2-oxoglutarate dehydrogenase inhibitor, OdhI, involved in glutamate production in Corynebacterium glutamicum

2017 ◽  
Vol 81 (11) ◽  
pp. 2130-2138 ◽  
Author(s):  
Ayano Komine-Abe ◽  
Megumi Nagano-Shoji ◽  
Shosei Kubo ◽  
Hisashi Kawasaki ◽  
Minoru Yoshida ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Lysine Succinylation ◽  
Glutamate Production ◽  
Oxoglutarate Dehydrogenase

Characterization of lysine acetylation in the peripheral subunit-binding domain of the E2 subunit of the pyruvate dehydrogenase-2-oxoglutarate dehydrogenase hybrid complex from Corynebacterium glutamicum

2020 ◽  
Vol 85 (4) ◽  
pp. 874-881
Author(s):  
Ayano Komine-Abe ◽  
Naoko Kondo ◽  
Shosei Kubo ◽  
Hisashi Kawasaki ◽  
Makoto Nishiyama ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Pyruvate Dehydrogenase ◽  
Critical Role ◽  
Binding Domain ◽  
Lysine Acetylation ◽  
Hybrid Complex ◽  
Glutamate Production ◽  
Accumulation Kinetic ◽  
Oxoglutarate Dehydrogenase

ABSTRACT In Corynebacterium glutamicum, pyruvate dehydrogenase (PDH) and 2-oxoglutarate dehydrogenase (ODH) form a unique hybrid complex in which CgE1p and CgE1o are associated with the CgE2–CgE3 subcomplex. We analyzed the role of a lysine acetylation site in the peripheral subunit-binding domain of CgE2 in PDH and ODH functions. Acetylation-mimic substitution at Lys391 of CgE2 severely reduced the interaction of CgE2 with CgE1p and CgE3, but not with CgE1o, indicating the critical role of this residue in the assembly of CgE1p and CgE3 into the complex. It also suggested that Lys391 acetylation inhibited the binding of CgE1p and CgE3 to CgE2, thereby affecting PDH and ODH activities. Interestingly, the CgE2-K391R variant strain showed increased l-glutamate production and reduced pyruvate accumulation. Kinetic analysis suggested that the increased affinity of the K391R variant toward pyruvate might be advantageous for l-glutamate production.

scholarly journals Altered Metabolic Flux due to Deletion of odhA causes l-Glutamate Overproduction in Corynebacterium glutamicum

2006 ◽  
Vol 73 (4) ◽  
pp. 1308-1319 ◽  
Author(s):  
Yoko Asakura ◽  
Eiichiro Kimura ◽  
Yoshihiro Usuda ◽  
Yoshio Kawahara ◽  
Kazuhiko Matsui ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Corynebacterium Glutamicum ◽  
Metabolic Flux ◽  
Dry Weight ◽  
Wild Type ◽  
Glutamate Production ◽  
Glutamate Dehydrogenase Activity ◽  
Tween 40 ◽  
Oxoglutarate Dehydrogenase ◽  
The Relationship

ABSTRACT l-Glutamate overproduction in Corynebacterium glutamicum, a biotin auxotroph, is induced by biotin limitation or by treatment with certain fatty acid ester surfactants or with penicillin. We have analyzed the relationship between the inductions, 2-oxoglutarate dehydrogenase complex (ODHC) activity, and l-glutamate production. Here we show that a strain deleted for odhA and completely lacking ODHC activity produces l-glutamate as efficiently as the induced wild type (27.8 mmol/g [dry weight] of cells for the ohdA deletion strain compared with only 1.0 mmol/g [dry weight] of cells for the uninduced wild type). This level of production is achieved without any induction or alteration in the fatty acid composition of the cells, showing that l-glutamate overproduction can be caused by the change in metabolic flux alone. Interestingly, the l-glutamate productivity of the odhA-deleted strain is increased about 10% by each of the l-glutamate-producing inductions, showing that the change in metabolic flux resulting from the odhA deletion and the inductions have additive effects on l-glutamate overproduction. Tween 40 was indicated to induce drastic metabolic change leading to l-glutamate overproduction in the odhA-deleted strain. Furthermore, optimizing the metabolic flux from 2-oxoglutarate to l-glutamate by tuning glutamate dehydrogenase activity increased the l-glutamate production of the odhA-deleted strain.

Enhancing the supply of oxaloacetate for l-glutamate production by pyc overexpression in different Corynebacterium glutamicum

2013 ◽  
Vol 35 (6) ◽  
pp. 943-950 ◽  
Author(s):  
Xuan Guo ◽  
Jing Wang ◽  
Xixian Xie ◽  
Qingyang Xu ◽  
Chenglin Zhang ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Glutamate Production

scholarly journals The Iron Deficiency Response of Corynebacterium glutamicum and a Link to Thiamine Biosynthesis

2020 ◽  
Vol 86 (10) ◽  
Author(s):  
Andreas Küberl ◽  
Aliye Mengus-Kaya ◽  
Tino Polen ◽  
Michael Bott
Keyword(s):  
Corynebacterium Glutamicum ◽  
Iron Homeostasis ◽  
Essential Element ◽  
Iron Limitation ◽  
Thiamine Pyrophosphate ◽  
Exponential Growth Phase ◽  
Iron Starvation ◽  
Content Type ◽  
Oxoglutarate Dehydrogenase ◽  
Thiamine Biosynthesis

ABSTRACT The response to iron limitation of the Gram-positive soil bacterium Corynebacterium glutamicum was analyzed with respect to secreted metabolites, the transcriptome, and the proteome. During growth in glucose minimal medium, iron limitation caused a shift from lactate to pyruvate as the major secreted organic acid complemented by l-alanine and 2-oxoglutarate. Transcriptome and proteome analyses revealed that a pronounced iron starvation response governed by the transcriptional regulators DtxR and RipA was detectable in the late, but not in the early, exponential-growth phase. A link between iron starvation and thiamine pyrophosphate (TPP) biosynthesis was uncovered by the strong upregulation of thiC. As phosphomethylpyrimidine synthase (ThiC) contains an iron-sulfur cluster, limiting activities of the TPP-dependent pyruvate–2-oxoglutarate dehydrogenase supercomplex probably cause the excretion of pyruvate and 2-oxoglutarate. In line with this explanation, thiamine supplementation could strongly diminish the secretion of these acids. The upregulation of thiC and other genes involved in thiamine biosynthesis and transport is presumably due to TPP riboswitches present at the 5′ end of the corresponding operons. The results obtained in this study provide new insights into iron homeostasis in C. glutamicum and demonstrate that the metabolic consequences of iron limitation can be due to the iron dependency of coenzyme biosynthesis. IMPORTANCE Iron is an essential element for most organisms but causes problems due to poor solubility under oxic conditions and due to toxicity by catalyzing the formation of reactive oxygen species (ROS). Therefore, bacteria have evolved complex regulatory networks for iron homeostasis aiming at a sufficient iron supply while minimizing ROS formation. In our study, the responses of the actinobacterium Corynebacterium glutamicum to iron limitation were analyzed, resulting in a detailed view on the processes involved in iron homeostasis in this model organism. In particular, we provide evidence that iron limitation causes TPP deficiency, presumably due to insufficient activity of the iron-dependent phosphomethylpyrimidine synthase (ThiC). TPP deficiency was deduced from the upregulation of genes controlled by a TPP riboswitch and secretion of metabolites caused by insufficient activity of the TPP-dependent enzymes pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase. To our knowledge, the link between iron starvation and thiamine synthesis has not been elaborated previously.

scholarly journals Characterization of a Corynebacterium glutamicum Lactate Utilization Operon Induced during Temperature-Triggered Glutamate Production

2005 ◽  
Vol 71 (10) ◽  
pp. 5920-5928 ◽  
Author(s):  
Corinna Stansen ◽  
Davin Uy ◽  
Stephane Delaunay ◽  
Lothar Eggeling ◽  
Jean-Louis Goergen ◽  
...  
Keyword(s):  
Lactate Dehydrogenase ◽  
Carbon Source ◽  
Corynebacterium Glutamicum ◽  
Specific Activity ◽  
Temperature Shift ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Reverse Transcription Pcr ◽  
Glutamate Production ◽  
Lactate Utilization

ABSTRACT Gene expression changes of glutamate-producing Corynebacterium glutamicum were identified in transcriptome comparisons by DNA microarray analysis. During glutamate production induced by a temperature shift, C. glutamicum strain 2262 showed significantly higher mRNA levels of the NCgl2816 and NCgl2817 genes than its non-glutamate-producing derivative 2262NP. Reverse transcription-PCR analysis showed that the two genes together constitute an operon. NCgl2816 putatively codes for a lactate permease, while NCgl2817 was demonstrated to encode quinone-dependent l-lactate dehydrogenase, which was named LldD. C. glutamicum LldD displayed Michaelis-Menten kinetics for the substrate l-lactate with a Km of about 0.51 mM. The specific activity of LldD was about 10-fold higher during growth on l-lactate or on an l-lactate-glucose mixture than during growth on glucose, d-lactate, or pyruvate, while the specific activity of quinone-dependent d-lactate dehydrogenase differed little with the carbon source. RNA levels of NCgl2816 and lldD were about 18-fold higher during growth on l-lactate than on pyruvate. Disruption of the NCgl2816-lldD operon resulted in loss of the ability to utilize l-lactate as the sole carbon source. Expression of lldD restored l-lactate utilization, indicating that the function of the permease gene NCgl2816 is dispensable, while LldD is essential, for growth of C. glutamicum on l-lactate.

An improved temperature-triggered process for glutamate production with Corynebacterium glutamicum

1999 ◽  
Vol 25 (8-9) ◽  
pp. 762-768 ◽  
Author(s):  
S. Delaunay ◽  
P. Gourdon ◽  
P. Lapujade ◽  
E. Mailly ◽  
E. Oriol ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Glutamate Production

scholarly journals The E2 Domain of OdhA of Corynebacterium glutamicum Has Succinyltransferase Activity Dependent on Lipoyl Residues of the Acetyltransferase AceF

2010 ◽  
Vol 192 (19) ◽  
pp. 5203-5211 ◽  
Author(s):  
Melanie Hoffelder ◽  
Katharina Raasch ◽  
Jan van Ooyen ◽  
Lothar Eggeling
Keyword(s):  
Corynebacterium Glutamicum ◽  
Pyruvate Dehydrogenase ◽  
The Other ◽  
Central Metabolism ◽  
Mutant Proteins ◽  
Oxoglutarate Dehydrogenase ◽  
The Individual ◽  
Activity Dependent ◽  
Dihydrolipoyl Dehydrogenase ◽  
Mutant Cells

ABSTRACT Oxoglutarate dehydrogenase (ODH) and pyruvate dehydrogenase (PDH) complexes catalyze key reactions in central metabolism, and in Corynebacterium glutamicum there is indication of an unusual supercomplex consisting of AceE (E1), AceF (E2), and Lpd (E3) together with OdhA. OdhA is a fusion protein of additional E1 and E2 domains, and odhA orthologs are present in all Corynebacterineae, including, for instance, Mycobacterium tuberculosis. Here we show that deletion of any of the individual domains of OdhA in C. glutamicum resulted in loss of ODH activity, whereas PDH was still functional. On the other hand, deletion of AceF disabled both PDH activity and ODH activity as well, although isolated AceF protein had solely transacetylase activity and no transsuccinylase activity. Surprisingly, the isolated OdhA protein was inactive with 2-oxoglutarate as the substrate, but it gained transsuccinylase activity upon addition of dihydrolipoamide. Further enzymatic analysis of mutant proteins and mutant cells revealed that OdhA specifically catalyzes the E1 and E2 reaction to convert 2-oxoglutarate to succinyl-coenzyme A (CoA) but fully relies on the lipoyl residues provided by AceF involved in the reactions to convert pyruvate to acetyl-CoA. It therefore appears that in the putative supercomplex in C. glutamicum, in addition to dihydrolipoyl dehydrogenase E3, lipoyl domains are also shared, thus confirming the unique evolutionary position of bacteria such as C. glutamicum and M. tuberculosis.

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