The Calvin cycle enzyme pentose-5-phosphate 3-epimerase is encoded within the cfx operons of the chemoautotroph Alcaligenes eutrophus

Kusian, Bernhard; Yoo, Je-Geun; Bednarski, R; Bowien, Botho

doi:10.1128/jb.174.22.7337-7344.1992

Cited by 50 publications

(44 citation statements)

References 53 publications

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“…A more likely explanation for the surprising fact that X flavus G7 is able to grow on succinate is that the mutant phosphoglycerate kinase enzyme retains some residual activity in vivo which is lost during preparation of the cell extract. A characterization of E. (6,14,17,25,32,35,42,48). Since the cbb operon promoter is active only during autotrophic growth, additional genes encoding the latter enzymes are present outside the cbb operon and are expressed during heterotrophic growth (31,48 (45), plays a role in the regulation of the pgk gene.…”

Section: Methodsmentioning

confidence: 99%

The Calvin cycle enzyme phosphoglycerate kinase of Xanthobacter flavus required for autotrophic CO2 fixation is not encoded by the cbb operon

Meijer

1994

J Bacteriol

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During autotrophic growth of Xanthobacterflavus, energy derived from the oxidation of hydrogen methanol or formate is used to drive the assimilation of CO2 via the Calvin cycle. The genes encoding the Calvin cycle enzymes are organized in the cbb operon, which is expressed only during autotrophic growth. Although it has been established that the transcriptional activator CbbR is required for the expression of the cbb operon, it is unclear whether CbbR is the only factor contributing to the regulation of the cbb operon. This paper describes the isolation of X. flavus mutants which were affected in the regulation of the cbb operon. One of the mutant strains was subject to an enhanced repression of the cbb operon promoter by the gluconeogenic substrate succinate and in addition failed to grow autotrophically. (9,32,33). A second fructosebisphosphatase gene required for heterotrophic growth is constitutively expressed (31,33).Transcription of the cbb operon is dependent on a LysRtype activator encoded by cbbR which binds to a low-and high-affinity site in the cbb operon promoter (45). Physiological studies have shown that high-level expression of the cbb operon is dependent on the absence of multicarbon substrates and the presence of methanol, formate, or hydrogen (9, 33). Although it has been established that CbbR is important in the transduction of these signals to the transcription apparatus (45), it is unclear how this is accomplished and whether CbbR is the only factor involved. To gain further insight into the molecular mechanism for the regulation of the cbb operon, a method that allowed the isolation of X flavus mutants with altered regulation of the cbb operon promoter was developed.The cbb operon promoter is not active during heterotrophic * Phone: (31)50-632150. Electronic mail address: meyerwg@biol.rug.nl. growth of X. flavus on succinate; however, when X flavus harbors multiple copies of the cbb operon promoter a low level of cbb operon promoter activity is observed during growth on succinate (26,30,32). This property was exploited to isolate X flavus mutants that lacked cbb operon promoter activity under these conditions and in addition failed to grow autotrophically. This paper describes the characterization of one of these mutants, X flavus G7, which was shown to be defective in phosphoglycerate kinase. This enzyme, which catalyzes the phosphorylation of 3-phosphoglycerate to 1,3-diphosphoglycerate, is required both for glycolysis and gluconeogenesis and for CO2 fixation via the Calvin cycle. In this paper, evidence that in contrast with the fructosebisphosphatase isoenzymes, X flavus employs only a single phosphoglycerate kinase that is responsible for both these functions is presented. MATERIALS AND METHODSBacterial strains and plasmids. The bacterial strains and plasmids used in this study are listed in Table 1.Media and growth conditions. Escherichia coli strains were grown on Luria-Bertani medium at 37°C (41). X flavus strains were grown on yeast extract (0.8% [wt/vol]) or in minimal media (27)...

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Section: Methodsmentioning

confidence: 99%

The Calvin cycle enzyme phosphoglycerate kinase of Xanthobacter flavus required for autotrophic CO2 fixation is not encoded by the cbb operon

Meijer

1994

J Bacteriol

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“…The deduced amino acid sequence of epimerase from potato is 57.4% identical with an epimerase from the photosynthetic purple bacterium Alcaligenes eutrophus [5]. Moreover, the search in EMBL and SwissProt databases for homologous proteins revealed two proteins from E. coli and a partial cDNA from Arabidopsis thaliana (Fig.…”

Section: Resultsmentioning

confidence: 88%

“…3. Alignment of the deduced amino acid sequences of potato epimerase with the deduced amino acid sequences of an epimerase from Ah'aligenes euthropus [5], and two epimerases from E coli [6,7]. Identical residues are indicated by stars, dots mark homologous amino acid substitutions,…”

Section: Resultsmentioning

confidence: 99%

“…Amino acid sequences deduced from eDNA sequences were reported for an epimerase encoded by the CO2-fixing operon @v of the purple bacterium Alcaligenes eutrophus [5] and two epimerase proteins from E. coli [6,7]. Purified epimerases from erythrocytes [2], calf liver [3], and E. eutrophus [5] The novel sequence reported here has been deposited with the EMBL database and is available under accession number Z50098.…”

Section: Introductionmentioning

confidence: 99%

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Chloroplast pentose‐5‐phosphate 3‐epimerase from potato: cloning, cDNA sequence, and tissue‐specific enzyme accumulation

1995

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A cDNA clone encoding the chloroplast enzyme pentose-5-phosphate 3-epimerase (EC 5.1.3.1) in potato (Solanum tuberosum) was isolated and sequenced. The deduced sequence of 235 amino acids is similar to protein sequences of bacterial epimerases. Northern blot analysis showed the highest level of epimerase mRNA expression in potato leaves, whereas it was low in roots, tubers, and stems. Epimerase protein is mulated only in plant tissues possessing chloroplasts, i.e. in land to a lesser extent in stem. In contrast, transketolase, a sequential enzyme of epimerase in the reductive and oxidative pentose phosphate cycle, is accumulated in all plant tissues.

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“…The remaining cbb genes, cbbE and cbbR, were sequenced entirely, on both strands, and like the preceding three genes, cbbR was promptly identified by the high degree of sequence similarity to cbbR genes from C. vinosum (48), A. eutrophus (54), and R. sphaeroides (18). The cbbE gene, on the other hand, coding for pentose-5-phosphate 3-epimerase, was not identified until a similar sequence in A. eutrophus was identified by Kusian et al (30). Thus, all the sequence upstream of cbbM on the XhoI fragment was assigned a coding function.…”

Section: G V L P H R M I E M S S N E T I K Qmentioning

confidence: 99%

Complementation analysis and regulation of CO2 fixation gene expression in a ribulose 1,5-bisphosphate carboxylase-oxygenase deletion strain of Rhodospirillum rubrum

Falcone

Tabita

1993

J Bacteriol

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A ribulose 1,5-bisphosphate carboxylase-oxygenase (RubisCO) deletion strain ofRhodospirillum rubrum that was incapable of photolithoautotrophic growth was constructed. Photoheterotrophic growth, however, was possible for the R. rubrum RubisCO deletion strain when oxidized carbon compounds such as malate were supplied. The R. rubrum RubisCO-deficient strain was not complemented to photolithoautotrophic growth by various R. rubrum DNA fragments that contain the gene encoding RubisCO, cbbM. When the R. rubrum cbbM deletion strain harbored plasmids containing R. rubrum DNA inserts with at least 2.0 kb preceding the translational start site of the cbbM gene, RubisCO activity and RubisCO antigen were detected. Lack of RubisCO expression was therefore not the cause for the failure to complement the cbbM mutant strain. Interestingly, DNA fragments encoding either of two complete Calvin-Benson-Bassham C02 fixation (cbb) gene operons from Rhodobacter sphaeroides were able to complement the R. rubrum RubisCO deletion strain to photolithoautotrophic growth. The same R. rubrum DNA fragments that failed to complement the R. rubrum cbbM deletion strain successfully complemented the RubisCO deletion strain of R. sphaeroides, pointing to distinct differences in the regulation of metabolism and the genetics of photolithoautotrophic growth in these two organisms. A number of cbb genes were identified by nucleotide sequence analysis of the region upstream of cbbM. Included among these was an open reading frame encoding a cbbR gene showing a high degree of sequence similarity to known lysR-type CO2 fixation transcriptional activator genes. The placement and orientation of the cbbR transcriptional regulator gene in R. rubrum are unique.

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The Calvin cycle enzyme pentose-5-phosphate 3-epimerase is encoded within the cfx operons of the chemoautotroph Alcaligenes eutrophus

Cited by 50 publications

References 53 publications

The Calvin cycle enzyme phosphoglycerate kinase of Xanthobacter flavus required for autotrophic CO2 fixation is not encoded by the cbb operon

The Calvin cycle enzyme phosphoglycerate kinase of Xanthobacter flavus required for autotrophic CO2 fixation is not encoded by the cbb operon

Chloroplast pentose‐5‐phosphate 3‐epimerase from potato: cloning, cDNA sequence, and tissue‐specific enzyme accumulation

Complementation analysis and regulation of CO2 fixation gene expression in a ribulose 1,5-bisphosphate carboxylase-oxygenase deletion strain of Rhodospirillum rubrum

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