• Korean Journal of Microbiology
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• v.28 no.3
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• pp.268-273
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• 1990

In order to elucidate the acid tolerance mechanism of Clostridium acetobutylicum against organic acid, the maintenance energy with added butyrate at different pH was determined. Maintenance coeffecient in acidogenic chemostat was higher at pH 6.5 than at pH 5.5 showing that this organism is an acidophile. The addition of butyrate at pH 5.5 and different dilution rate caused linear decrease of the cell concentration though $Y_{ATP}$ did not decrease with increasing undissociated organic acid. $Y_{ATP}$ decreased by increasing the concentration of undissociated organic acid at pH 5.0 by the addition of butyrate. From these results it is hypothesized that the ATP conaumption for pH stat of acidophile C. acetobutylicum is increased at the circumstance with over 30mM of undissociated organic acid.cid.

• Journal of Microbiology and Biotechnology
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• v.26 no.4
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• pp.725-729
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• 2016

A novel genome-engineering tool in Clostridium acetobutylicum was developed based on single-crossover homologous recombination. A small-sized non-replicable plasmid, pHKO1, was designed for efficient integration into the C. acetobutylicum genome. The integrated pHKO1 plasmid backbone, which included an antibiotic resistance gene, can be excised in vivo by Flp recombinase, leaving a single flippase recognition target sequence in the middle of the targeted gene. Since the pSHL-FLP plasmid, the carrier of the Flp recombinase gene, employed the segregationally unstable pAMβ1 replicon, the plasmid was rapidly cured from the mutant C. acetobutylicum. Consequently, our method makes it easier to engineer C. acetobutylicum.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1702-1708
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• 2015

We have developed a new shuttle plasmid, designated as pLK1-MCS that can replicate in both Clostridium acetobutylicum and Escherichia coli, by combining the pUB110 and pUC19 plasmids. Plasmid pLK1-MCS replicated more stably than previously reported plasmids containing either the pIM13 or the pAMβ1 replicon in the absence of antibiotic selective pressure. The transfer frequency of pLK1-MCS into C. acetobutylicum was similar to the transfer frequency of other shuttle plasmids. We complemented C. acetobutylicum ML1 (that does not produce solvents such as acetone, butanol, and ethanol owing to loss of the megaplasmid pSOL1 harboring the adhE1-ctfAB-adc operon) by introducing pLK1-MCS carrying the adhE1-ctfAB-adc operon into C. acetobutylicum ML1. The transformed cells were able to resume anaerobic solvent production, indicating that the new shuttle plasmid has the potential for practical use in microbial biotechnology.

• The Microorganisms and Industry
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• v.18 no.3
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• pp.2-9
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• 1992

이 논문에서는 대사공학에의 응용에 필수적이며 또한 그 자체의 기술이 학문적으로 상당히 관심을 끄는 C. acetobutylicum에서의 primary metabolic gene cloning에 대하여 정리해 보고자 한다. 우선 C. acetobutylicum의 primary metabolism과 일반적인 대사 조절에 대하여 간략히 살펴보고 이에 관여한 효소들과 gene cloning에 대하여 기술하고자 한다.

• KSBB Journal
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• v.8 no.1
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• pp.1-9
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• 1993

The acetone-butanol fermentation by C. acetobutylicum has gained increasing attention for the following reasons. First, the finite supply of petrochemical resources, combined with increasing concern over global environmental effects and the unstable nature of the price of petroleum has renewed interest in the development of fermentation technology that allows utilzation of biomass wastes for the production of alcohol. Second, it serves as excellent model system for understading the regulation and molecular biology of tightly regulated complex primary metabolism, and for applications of metabolic engineering. In this review various aspects of acetone-butanol fermentation by C. acetobutylicm including strain and fermentation characteristics, enzyme regulation, and solvent formation mechanism, and product recovery and summarized.

• Korean Chemical Engineering Research
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• v.51 no.2
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• pp.226-232
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• 2013

To produce biobutanol, fermentation processes using clostridia that mainly produce acetone, butanol and ethanol are used. In this work, a dynamic model describing the metabolic reactions in an acetone-butanol-ethanol (ABE)-producing clostridium, Clostridium acetobutylicum ATCC824, was proposed. To estimate the 58 kinetic parameters of the metabolic network model with experimental data obtained from a batch fermentor, we used an efficient optimization method combining a genetic algorithm and the Levenberg-Marquardt method because of the complexity of the metabolism of the clostridium. For the verification of the determined parameters, the developed metabolic model was evaluated by experiments where genetically modified clostridium was used and the initial concentration of glucose was changed. Consequently, we found that the developed kinetic model for the metabolic network was considered to describe the dynamic metabolic state of the clostridium sufficiently. Thus, this dynamic model for the metabolic reactions will contribute to designing the clostridium as well as the fermentor for higher productivity.

• Journal of Microbiology and Biotechnology
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• v.2 no.4
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• pp.268-272
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• 1992

Alcohol dehydrogenase activity of Clostridium acetobutylicum ATCC 4259 was studied for its specificity against substrates in acidogenic and solventogenic cultures. The bacterium reduces propionate, valerate and caproate added to the medium to the corresponding alcohols. Acetaldehyde, propionaldehyde, butyraldhyde, pentanal, and hexanal were used as the substrates by alcohol dehydrogenase, and all were reduced to the corresponding alcohols with varying affinities and reaction velocities. Acetaldehyde showed the lowest affinity and lowest velocity while the other aldehydes showed similar $K_m\;and\;V_max$ values. NADPH was used as the electron donor for the reduction of aldehydes. Alcohol dehydrogenase activity was low in acidogenic culture, and high in solventogenic culture.

• Journal of Microbiology and Biotechnology
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• v.31 no.10
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• pp.1393-1400
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• 2021

Acetone-butanol-ethanol (ABE) fermentation by the anaerobic bacterium Clostridium acetobutylicum has been considered a promising process of industrial biofuel production. Phosphotransbutyrylase (phosphate butyryltransferase, PTB) plays a crucial role in butyrate metabolism by catalyzing the reversible conversion of butyryl-CoA into butyryl phosphate. Here, we report the crystal structure of PTB from the Clostridial host for ABE fermentation, C. acetobutylicum, (CaPTB) at a 2.9 Å resolution. The overall structure of the CaPTB monomer is quite similar to those of other acyltransferases, with some regional structural differences. The monomeric structure of CaPTB consists of two distinct domains, the N- and C-terminal domains. The active site cleft was formed at the interface between the two domains. Interestingly, the crystal structure of CaPTB contained eight molecules per asymmetric unit, forming an octamer, and the size-exclusion chromatography experiment also suggested that the enzyme exists as an octamer in solution. The structural analysis of CaPTB identifies the substrate binding mode of the enzyme and comparisons with other acyltransferase structures lead us to speculate that the enzyme undergoes a conformational change upon binding of its substrate.

• KSBB Journal
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• v.5 no.1
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• pp.9-17
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• 1990

The growth and fermentation profiles of Clostridium acetobutylicum KCTC 1037 were examined in batch and continuous modes with pH variation and phosphate limitation. Clostridium acetobutylicum KCTC 10 37 grew better at pH 4.5 than at pH 5.5 or 6.5. Acetate and butyrate were produced at pH 5.5, whereas culture at pH 4.5 produced acetone and butanol. Solvent production was increased by the phosphate limitation in a batch culture, but in a phosphate-limited continuous culture for 400 hours steady-state solventogenesis was not observed. The induction and maintenance of solventogenesis presumably require not only acidic condition or phosphate limitation but also favourable bioenergetic condition.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.236-242
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• 2017

The conversion of marine biomass to renewable energy has been considered an alternative to fossil fuels. Butanol, in particular, can be used directly as a fuel. In this experiment, the brown alga Undaria pinnatifida was selected as a biomass for biobutanol production. Hyper thermal (HT) acid hydrolysis was used as an acid hydrolysis method to produce monosaccharides. The optimal pretreatment conditions for U. pinnatifida were determined as slurry with 10% (w/v) U. pinnatifida content and 270 mM $H_2SO_4$, and heating at $160^{\circ}C$ for 7.5 min. Enzymatic saccharification was carried out with Celluclast 1.5 L, Viscozyme L, and Ultraflo Max. The optimal saccharification condition was 12 U/ml Viscozyme L. Fermentations were carried out for the production of acetone, butanol, and ethanol by Clostridium acetobutylicum KCTC 1724, Clostridium beijerinckii KCTC 1785, and Clostridium tyrobutyricum KCTC 5387. The fermentations were carried out using a pH-control. The optimal ABE fermentation condition determined using C. acetobutylicum KCTC 1724 adapted to 160 g/l mannitol. An ABE concentration of 9.05 g/l (0.99 g/l acetone, 5.62 g/l butanol, 2.44 g/l ethanol) was obtained by the consumption of 24.14 g/l monosaccharide with $Y_{ABE}$ of 0.37 in pH 5.0.