Sequence Analysis of theatpOperon ofClostridium acetobutylicumDSM 792 Enioding theF0F1ATP Synthase

Externbrink, Thorsten; Hujer, Sandra; Winzer, Klaus; Dürre, Peter

doi:10.3109/10425170009033977

Cited by 5 publications

(8 citation statements)

References 19 publications

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“…+ and À in ''Gene included in microarray'' indicate relative transcript levels detected in microarray experiments: ++, very high expression; +, moderately high expression; À moderately low expression; ÀÀ very low expression. When the transcript levels of a gene was examined in both microarray experiments, the relative expression levels for both are indicated and separated by comma e Gene is included on microarray and transcript expression profile examined in wild type 824 strain compared to SKO1 strain [42] f Exp, exponential phase, Tran, Transition, Stat, stationary phase g +/À is up-/downregulated, respectively; NC is no change, thus approximately constitutive levels of the overall protein accumulation trend in C. acetobutylicum 824 (pIMP1) h References for known operons: [6,12,28] i Number in front of 0A or R0A indicates the number of non-overlapping boxes present, if no number is indicated, a single box is present j Each abundance refers to one variant of the protein identified k Based on gene clustering with other genes of similar function in the same orientation l May be demonstrated or putative operon structure …”

Section: Resultsmentioning

confidence: 99%

“…Tpi from various clostridia have different heat stabilities [13], and the Tpi from the three examined clostridia all had the same MW of 53 kDa as dimers, but with different pIs [41]. ATP synthase catalyzes the formation of ATP from ADP and inorganic phosphate using a proton or sodium ion gradient [12]. AtpD abundance decreases during exponential phase and is undetectable by late solventogenesis.…”

Section: Downregulated Proteins Observedmentioning

confidence: 99%

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Proteome analysis and comparison of Clostridium acetobutylicum ATCC 824 and Spo0A strain variants

Sullivan

Bennett

2005

J IND MICROBIOL BIOTECHNOL

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The proteomic profiles of several Clostridium acetobutylicum strains were compared by two-dimensional gel electrophoresis and mass spectroscopy. The proteomic profile of C. acetobutylicum wild type strain ATCC 824 with and without a commonly used control plasmid and with a spo0A overexpression plasmid pMSPOA was compared. A total of 2,081 protein spots were analyzed; 23 proteins were chosen to be identified of which 18 were unique and 5 were proteins located in more than one location. The proteins identified were classified into heat shock stress response, acid and solvent formation, and transcription and translation proteins. Spo0A was identified and its protein expression was confirmed to be absent in the spo0A knockout SKO1 strain as expected, as was the protein Adc, which is known to be regulated by Spo0A. The expression of six proteins was not detected in strain SKO1 indicating these proteins require Spo0A. Spo0A overexpression affected the abundance of proteins involved in glycolysis, translation, heat shock stress response, and energy production. Two features were identified: five of the 23 proteins identified were located in more than one position and clusters of protein spots resembled fingers of a straightened hand. Normally a protein localizes to only one spot on the gel; localization of a protein to more than one spot is indicative of post-translational modifications, suggesting that such modification of proteins may be a more prevalent mechanism in C. acetobutylicum than previously thought. The clusters of protein spots resembling fingers of a straightened hand were in the acidic high molecular weight areas. Two such protein spots were identified as variants of the same protein, GroEL.

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Section: Resultsmentioning

confidence: 99%

Section: Downregulated Proteins Observedmentioning

confidence: 99%

Proteome analysis and comparison of Clostridium acetobutylicum ATCC 824 and Spo0A strain variants

Sullivan

Bennett

2005

J IND MICROBIOL BIOTECHNOL

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“…Northern blotting and primer extension analysis were performed as described recently [13,14]. Hybridization probes for Northern experiments were synthesized by PCR [13,14] using primer pairs adhEl (5'-GCCCTTAAGAGTGGTGCCCC-3') Figure 1 shows the autoradiographs of second-dimensional gels after separation of radioactively labeled cellfree extracts from acidogenic and solventogenic C. aceto-…”

Section: Rna Analysesmentioning

confidence: 99%

Changes in protein synthesis and identification of proteins specifically induced during solventogenesis in Clostridium acetobutylicum

et al. 2002

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At the end of the exponential growth phase the Gram-positive bacterium Clostridium acetobutylicum performs a metabolic switch from classical sugar fermentation accompanied by the production of acetate and butyrate to reinternalization and oxidation of these acids to acetone and butanol. Protein synthesis in acidogenic and solventogenic C. acetobutylicum cells was compared by two-dimensional gel electrophoresis of radioactively labeled proteins. The results show that the switch from acid to solvent production is accompanied by dramatic changes in the protein pattern. During solventogenesis, the synthesis of 52 proteins out of 130 analyzed was increased more than twofold, the synthesis of 34 proteins decreased to less than half as compared with synthesis in acidogenic cells. The changes in protein synthesis were generally reflected by changes in the abundance of the respective proteins, as determined from quantitative analysis of silver-stained second-dimensional gels. Nine proteins induced during solventogenesis were identified by N-terminal microsequencing. One of these proteins, the acetoacetate decarboxylase, is directly involved in solventogenesis. Other proteins synthesized in higher amounts during solventogenesis were three general stress proteins (DnaK, GroEL, Hsp 18), two enzymes involved in serine biosynthesis (serine aminotransferase and 3-phosphoglycerate dehydrogenase) as well as a seryl-tRNA synthetase. mRNA analysis provided evidence that the latter three are encoded by genes organized in an operon and are transcriptionally induced at the onset of solventogenesis. The proteins acetoacetate decarboxylase and Hsp 18 occurred in two variants, indicating possible covalent modification of these proteins.

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“…The reason for this biphasic fermentation is closely related to the energy and redox metabolism in C. acetobutylicum (Figure 1) (Jang et al, 2014a;Lütke-Eversloh, 2014). In these bacteria, ATP is primarily produced from glucose through glycolysis (Externbrink et al, 2000). During the initial growth phase in C. acetobutylicum, additional ATP is produced through substrate-level phosphorylation, which is coupled to the production of acetate and butyrate (Externbrink et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

“…In these bacteria, ATP is primarily produced from glucose through glycolysis (Externbrink et al, 2000). During the initial growth phase in C. acetobutylicum, additional ATP is produced through substrate-level phosphorylation, which is coupled to the production of acetate and butyrate (Externbrink et al, 2000). At that time, to regenerate NAD + , NADH could be oxidized via not only two enzymes 3-hydroxybutyryl-CoA dehydrogenase (HBD) and butyryl-CoA dehydrogenase (BCD) responsible for butyrate formation, but also hydrogenase (HYD) coupled with ferredoxin oxidoreductase (PFOR; Figure 1) (Du et al, 2021;Jiang et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Clostridium acetobutylicum atpG-Knockdown Mutants Increase Extracellular pH in Batch Cultures

Jang

Seong

Kwon

et al. 2021

Front. Bioeng. Biotechnol.

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ATPase, a key enzyme involved in energy metabolism, has not yet been well studied in Clostridium acetobutylicum. Here, we knocked down the atpG gene encoding the ATPase gamma subunit in C. acetobutylicum ATCC 824 using a mobile group II intron system and analyzed the physiological characteristics of the atpG gene knockdown mutant, 824-2866KD. Properties investigated included cell growth, glucose consumption, production of major metabolites, and extracellular pH. Interestingly, in 2-L batch fermentations, 824-2866KD showed no significant difference in metabolite biosynthesis or cell growth compared with the parent ATCC 824. However, the pH value in 824-2866KD cultures at the late stage of the solventogenic phase was abnormally high (pH 6.12), compared with that obtained routinely in the culture of ATCC 824 (pH 5.74). This phenomenon was also observed in batch cultures of another C. acetobutylicum, BEKW-2866KD, an atpG-knockdown and pta-buk double-knockout mutant. The findings reported in this study suggested that ATPase is relatively minor than acid-forming pathway in ATP metabolism in C. acetobutylicum.

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Sequence Analysis of theatpOperon ofClostridium acetobutylicumDSM 792 Enioding theF₀F₁ATP Synthase

Cited by 5 publications

References 19 publications

Proteome analysis and comparison of Clostridium acetobutylicum ATCC 824 and Spo0A strain variants

Proteome analysis and comparison of Clostridium acetobutylicum ATCC 824 and Spo0A strain variants

Changes in protein synthesis and identification of proteins specifically induced during solventogenesis in Clostridium acetobutylicum

Clostridium acetobutylicum atpG-Knockdown Mutants Increase Extracellular pH in Batch Cultures

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