Acetone-butanol fermentation by mutants selected for resistance to acetate and butyrate halogen analogues

Junelles, Anne-Marie; Janati‐Idrissi, Rachid; Kanouni, A. El; Petitdemange, H.

doi:10.1007/bf01024559

Cited by 15 publications

(4 citation statements)

References 8 publications

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“…This finding and the altered morphology of the wild-type strain on low levels of the analogs may suggest that an alteration in surface structure or transport properties could be involved. The 2-BB mutant R exhibits a low production of acetone, as was found for a 2-BB mutant in the earlier report (10). From assays of enzymes of the solvent pathway in this strain, it appears that CoA-transferase is the only solvent pathway enzyme present in significantly reduced quantity.…”

Section: Discussionsupporting

confidence: 77%

“…Inefficient plating and the difficulty of obtaining highly efficient mutagenesis with most common C. acetobutylicum strains have been two restraints on the advance of genetics of this anaerobe. A few successful approaches for obtaining useful solvent pathway mutants have been reported, including the use of alcohol or acid analogs which are toxic upon enzymatic conversion by the cell (4,10,16,19). Growth in the presence of these analogs has given rise to mutants with reduced butanol levels which produce butyraldehyde in addition to normal amounts of acetone (16).…”

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confidence: 99%

“…A second method (10) was to plate an exponential-phase culture directly on selective medium and place a filter paper disk, impregnated with 75 .l of a saturated solution of NMNG, on the center of the plate.…”

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confidence: 99%

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Isolation and Characterization of Mutants of Clostridium acetobutylicum ATCC 824 Deficient in Acetoacetyl-Coenzyme A:Acetate/Butyrate:Coenzyme A-Transferase (EC 2.8.3.9) and in Other Solvent Pathway Enzymes

Clark

Bennett

Rudolph

1989

Appl Environ Microbiol

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Mutants of Clostridium acetobutylicum ATCC 824 exhibiting resistance to 2-bromobutyrate or rifampin were isolated after nitrosoguanidine treatment. Mutants were screened for solvent production by using an automated alcohol test system. Isolates were analyzed for levels of butanol, ethanol, acetone, butyrate, acetate, and acetoin in stationary-phase batch cultures. The specific activities of NADHand NADPH-dependent butanol dehydrogenase and butyraldehyde dehydrogenase as well as those of acetoacetyl-coenzyme A:acetate/butyrate: coenzyme A-transferase (butyrate-acetoacetate coenzyme A-transferase [EC 2.8.3.91) (CoA-transferase), butyrate kinase, and phosphotransbutyrylase were measured at the onset of stationary phase. Rifampinresistant strain D10 and 2-bromobutyrate mutant R were found to be deficient in only CoA-transferase, while several other mutants exhibited reduced butyraldehyde dehydrogenase and butanol dehydrogenase activities as well. The colony morphology of 2-bromobutyrate mutant R was similar to that of the parent on RCM medium; however, it had about 1/10 the level of CoA-transferase and increased levels of butanol dehydrogenase and butyraldehyde dehydrogenase. A nonsporulating, spontaneously derived degenerated strain exhibited reduced levels of butyraldehyde dehydrogenase, butanol, dehydrogenase, and CoA-transferase compared with those of the original strain. When C. acetobutylicum ATCC 824 was grown on medium containing low levels of 2-bromobutyrate, an altered colony morphology was observed. Not all strains resistant to 2-bromobutyrate (12 mM) were non-solvent-producing strains.

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Section: Discussionsupporting

confidence: 77%

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confidence: 99%

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Isolation and Characterization of Mutants of Clostridium acetobutylicum ATCC 824 Deficient in Acetoacetyl-Coenzyme A:Acetate/Butyrate:Coenzyme A-Transferase (EC 2.8.3.9) and in Other Solvent Pathway Enzymes

Clark

Bennett

Rudolph

1989

Appl Environ Microbiol

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“…Physical mapping of the genome demonstrated that this strain has a 4-Mb chromosome with 11 ribosomal operons (9) and harbors a large plasmid, about 200 kb in size, which carries the genes involved in solvent formation, hence the name pSOL1 (10). Much work has been done to elucidate the metabolic pathways by which solvents are produced and to isolate solvent-tolerant or solvent-overproducing strains (8,21,35,62,69,71,80). Genetic systems have been developed that allow genes to be manipulated in C. acetobutylicum ATCC824 and related organisms (25,(48)(49)(50)(51)(52)84), and these have been used to develop modified strains with altered solventogenic properties (25,28,54,60).…”

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Genome Sequence and Comparative Analysis of the Solvent-Producing BacteriumClostridium acetobutylicum

et al. 2001

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The genome sequence of the solvent-producing bacterium Clostridium acetobutylicum ATCC 824 has been determined by the shotgun approach. The genome consists of a 3.94-Mb chromosome and a 192-kb megaplasmid that contains the majority of genes responsible for solvent production. Comparison of C. acetobutylicum to Bacillus subtilis reveals significant local conservation of gene order, which has not been seen in comparisons of other genomes with similar, or, in some cases closer, phylogenetic proximity. This conservation allows the prediction of many previously undetected operons in both bacteria. However, the C. acetobutylicum genome also contains a significant number of predicted operons that are shared with distantly related bacteria and archaea but not with B. subtilis. Phylogenetic analysis is compatible with the dissemination of such operons by horizontal transfer. The enzymes of the solventogenesis pathway and of the cellulosome of C. acetobutylicum comprise a new set of metabolic capacities not previously represented in the collection of complete genomes. These enzymes show a complex pattern of evolutionary affinities, emphasizing the role of lateral gene exchange in the evolution of the unique metabolic profile of the bacterium. Many of the sporulation genes identified in B. subtilis are missing in C. acetobutylicum, which suggests major differences in the sporulation process. Thus, comparative analysis reveals both significant conservation of the genome organization and pronounced differences in many systems that reflect unique adaptive strategies of the two gram-positive bacteria.

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Analysis of Degenerate Variants ofClostridium acetobutylicumATCC 824

et al. 1996

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Acetone-butanol fermentation by mutants selected for resistance to acetate and butyrate halogen analogues

Cited by 15 publications

References 8 publications

Isolation and Characterization of Mutants of Clostridium acetobutylicum ATCC 824 Deficient in Acetoacetyl-Coenzyme A:Acetate/Butyrate:Coenzyme A-Transferase (EC 2.8.3.9) and in Other Solvent Pathway Enzymes

Isolation and Characterization of Mutants of Clostridium acetobutylicum ATCC 824 Deficient in Acetoacetyl-Coenzyme A:Acetate/Butyrate:Coenzyme A-Transferase (EC 2.8.3.9) and in Other Solvent Pathway Enzymes

Genome Sequence and Comparative Analysis of the Solvent-Producing BacteriumClostridium acetobutylicum

Analysis of Degenerate Variants ofClostridium acetobutylicumATCC 824

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