Characterization of recombinant strains of theClostridium acetobutylicum butyrate kinase inactivation mutant: Need for new phenomenological models for solventogenesis and butanol inhibition?

Harris, Latonia M.; Desai, Ruchir P.; Welker, N. E.; Papoutsakis, Eleftherios T.

doi:10.1002/(sici)1097-0290(20000105)67:1<1::aid-bit1>3.0.co;2-g

Cited by 158 publications

(29 citation statements)

References 15 publications

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“…This would support our current observation that acetate production is inhibited at a lower concentration (50 mM) in the AK strain than for the wild type, which accumulates more than 80 mM of acetate in CM1 medium under the same conditions. This is similar to strain ATCC 824 that accumulates up to 71 mM in CGM medium (Harris et al 2000). …”

Section: Discussionsupporting

confidence: 79%

“…The reduced lactate production levels of the AK mutant, relative to those by the wild-type, are not in line with this expectation. The wild-type C. acetobutylicum WUR strain showed (high) lactate production levels on CGM in contrast to strain ATCC 824 which produced none under similar conditions (Desai and Papoutsakis 1999; Harris et al 2000). This dissimilarity could be due to a different regulation of gene expression or different enzymatic properties of the iso-enzymes involved in lactate formation in both strains.…”

Section: Discussionmentioning

confidence: 96%

“…The inactivation of the butyrate kinase ( buk1 ) gene of C. acetobutylicum has previously been shown to result in reduced butyrate formation, increased peak acetate levels and increased butanol and ethanol production (Harris et al 2000). Acetone is one of the major by-products of butanol production as it is formed as a consequence of the uptake of acids from the medium.…”

Section: Discussionmentioning

confidence: 99%

“…In this substrate level phosphorylation reaction, the phosphate group of acetyl phosphate is transferred to ADP, generating ATP and acetate. In addition, we were interested to see if solvent formation would be similarly beneficially affected, as reported for buk1 gene disruption (Harris et al 2000). …”

Section: Discussionmentioning

confidence: 99%

“…To date, only two gene knockouts (phosphotransacetylase, pta ; butyrate kinase, buk1 ) (Green et al 1996; Shao et al 2007) and five asRNA constructs (acetoacetyl-CoA:acetate/butyrate:CoA-transferase, ctfA and ctfB ; acetoacetate decarboxylase, adc ; phosphotransbutyrylase, ptb ; buk1 ) (Desai and Papoutsakis 1999; Tummala et al 2003a, b) pertaining directly to the reduction of by-product formation in C. acetobutylicum ATCC 824 have been reported. Inactivation of buk1 resulted in a 42% increase in butanol levels (Harris et al 2000) while inactivation of the pta , involved in acetate formation, did not show improved solvent production (WT A-B-E production, 72:133:13 mM vs. 79:131:11 for the pta − mutant) (Green et al 1996). …”

Section: Introductionmentioning

confidence: 99%

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Disruption of the acetate kinase (ack) gene of Clostridium acetobutylicum results in delayed acetate production

Kuit

Minton

López-Contreras

et al. 2012

Appl Microbiol Biotechnol

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In microorganisms, the enzyme acetate kinase (AK) catalyses the formation of ATP from ADP by de-phosphorylation of acetyl phosphate into acetic acid. A mutant strain of Clostridium acetobutylicum lacking acetate kinase activity is expected to have reduced acetate and acetone production compared to the wild type. In this work, a C. acetobutylicum mutant strain with a selectively disrupted ack gene, encoding AK, was constructed and genetically and physiologically characterized. The ack− strain showed a reduction in acetate kinase activity of more than 97% compared to the wild type. The fermentation profiles of the ack− and wild-type strain were compared using two different fermentation media, CGM and CM1. The latter contains acetate and has a higher iron and magnesium content than CGM. In general, fermentations by the mutant strain showed a clear shift in the timing of peak acetate production relative to butyrate and had increased acid uptake after the onset of solvent formation. Specifically, in acetate containing CM1 medium, acetate production was reduced by more than 80% compared to the wild type under the same conditions, but both strains produced similar final amounts of solvents. Fermentations in CGM showed similar peak acetate and butyrate levels, but increased acetoin (60%), ethanol (63%) and butanol (16%) production and reduced lactate (−50%) formation by the mutant compared to the wild type. These findings are in agreement with the proposed regulatory function of butyryl phosphate as opposed to acetyl phosphate in the metabolic switch of solventogenic clostridia.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-011-3848-4) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 79%

Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Disruption of the acetate kinase (ack) gene of Clostridium acetobutylicum results in delayed acetate production

Kuit

Minton

López-Contreras

et al. 2012

Appl Microbiol Biotechnol

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Biological Production of Butanol and Higher Alcohols

Zhao¹,

Lu²,

Chen³

et al. 2013

Bioprocessing Technologies in Biorefinery for Sustainable Production of Fuels, Chemicals, and Polymers

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Metabolic engineering of clostridia for the production of chemicals

Cho

Jang

Moon

et al. 2014

Biofuels Bioprod Bioref

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There have recently been signifi cant advances in bio-based production of chemicals from renewable resources. Microorganisms belonging to the genus Clostridium have been considered as one of the promising hosts for the production of desired chemicals of wide industrial use. Clostridium strains have capability to utilize diverse carbon sources, including C5 and C6 substrates, which thus allows production of chemicals from inexpensive and abundant biomass such as corn stover, straw, and woody waste. In addition, Clostridium strains naturally produce various chemicals, such as acetic acid, butyric acid, ethanol, isopropanol, butanol, 1,3-propanediol, 2,3-butanediol, and acetone. Recently, several important strategies for the metabolic engineering of Clostridium have been developed not only for the enhanced production of these natural products and but also for the production of non-natural isobutanol production. Here, we review the strategies employed for the development of metabolically engineered Clostridium strains for the production of such chemicals and provide future perspectives.

show abstract

Characterization of recombinant strains of theClostridium acetobutylicum butyrate kinase inactivation mutant: Need for new phenomenological models for solventogenesis and butanol inhibition?

Cited by 158 publications

References 15 publications

Disruption of the acetate kinase (ack) gene of Clostridium acetobutylicum results in delayed acetate production

Disruption of the acetate kinase (ack) gene of Clostridium acetobutylicum results in delayed acetate production

Biological Production of Butanol and Higher Alcohols

Metabolic engineering of clostridia for the production of chemicals

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