• Journal of Microbiology and Biotechnology
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• 제32권8호
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• pp.1047-1053
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• 2022

When ptsG, a glucose-specific phosphotransferase system (PTS) component, is deleted in Escherichia coli, growth can be severely poor because of the lack of efficient glucose transport. We discovered a new PTS transport system that could transport glucose through the growth-coupled experimental evolution of ptsG-deficient E. coli C strain under anaerobic conditions. Genome sequencing revealed mutations in agaR, which encodes a repressor of N-acetylgalactosamine (Aga) PTS expression in evolved progeny strains. RT-qPCR analysis showed that the expression of Aga PTS gene increased because of the loss-of-function of agaR. We confirmed the efficient Aga PTS-mediated glucose uptake by genetic complementation and anaerobic fermentation. We discussed the discovery of new glucose transporter in terms of different genetic backgrounds of E. coli strains, and the relationship between the pattern of mixed-acids fermentation and glucose transport rate.

• 한국수소및신에너지학회논문집
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• 제19권2호
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• pp.145-155
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• 2008

This paper deals with an economic evaluation of hydrogen production by fermentation. We evaluate the economic feasibility of domestic hydrogen production by fermentation utilizing glucose and waste water sludge in terms of hydrogen production prices. In addition, we make some sensitivity analysis of hydrogen prices by changing the values of input factors such as the price of glucose, the capital cost of the hydrogen production system, and the hydrogen production yields. The estimated hydrogen prices of the two-step dark-light hydrogen production by fermentation utilizing glucose was $5,347won/kgH_2$, and the single-step hydrogen production by anaerobic fermentation utilizing waste water sludge was $4,255won/kgH_2$, respectively. It is expected that the hydrogen production price by anaerobic fermentation can be reduced if we produce methane or hydrogen utilizing by-products such as alcohols and organic acids, or the government imposes some legal regulations on the treatment of waste water sludge.

• Journal of Microbiology and Biotechnology
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• 제26권3호
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• pp.627-636
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• 2016

The causative agent of pandemic cholera, Vibrio cholerae, infects the anaerobic environment of the human intestine. Production of cholera toxin (CT), a major virulence factor of V. cholerae, is highly induced during anaerobic respiration with trimethylamine N-oxide (TMAO) as an alternative electron acceptor. However, the molecular mechanism of TMAO-stimulated CT production is not fully understood. Herein, we reveal that CT production during anaerobic TMAO respiration is affected by glucose fermentation. When the seventh pandemic V. cholerae O1 strain N16961 was grown with TMAO and additional glucose, CT production was markedly reduced. Furthermore, an N16961 Δcrp mutant, devoid of cyclic AMP receptor protein (CRP), was defective in CT production during growth by anaerobic TMAO respiration, further suggesting a role of glucose metabolism in regulating TMAO-mediated CT production. TMAO reductase activity was noticeably decreased when grown together with glucose or by mutation of the crp gene. A CRP binding region was identified in the promoter region of the torD gene, which encodes a structural subunit of the TMAO reductase. Gel shift assays further confirmed the binding of purified CRP to the torD promoter sequence. Together, our results suggest that the bacterial ability to respire using TMAO is controlled by CRP, whose activity is dependent on glucose availability. Our results reveal a novel mechanism for the regulation of major virulence factor production by V. cholerae under anaerobic growth conditions.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권1호
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• pp.54-57
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• 2003

H$_2$-producing bacteria were isolated from anaerobic granular sludge. Out of 72 colonies (36 grown under aerobic conditions and 36 under anaerobic conditions) arbitrarily chosen from the agar plate cultures of a Suspended sludge, 34 colonies (15 under aerobic conditions and 19 under anaerobic conditions) produced H2 under anaerobic conditions. Based on various biochemical tests and microscopic observations, they were classified into 13 groups and tentatively identified as follows: From aerobic isolates, Aeromonar spp. (7 strains), Pseudomonas spp. (3 strains), and Vibrio spp. (5 strains); from anaerobic isolates, Actinomyces spp. (11 Strains), Clostridium 5pp. (7 strains). and Porphyromonas sp. When glucose was used as the carbon substrate, all isolates showed a similar cell density and a H$_2$ production yield in the batch cultivations after 12 h (2.24-2.74 OD at 600 nm and 1.02-1.22 mol H$_2$/mol glucose, respectively). The major fermentation by-products were ethanol and acetate for the aerobic isolates, and ethanol, acetate and propionate for the anaerobic isolates. This study demonstrated that several H$_2$ producers in an anaerobic granular sludge exist En large proportions and their performance in terms of H$_2$ production is quite similar.

• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.532-538
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• 2008

For the newly isolated $H_2$-producing chemoheterotrophic bacterium Citrobacter amalonaticus Y19, anaerobic glucose metabolism was studied in batch cultivation at varying initial glucose concentrations (3.5-9.5 g/l). The carbon-mass and energy balances were determined and utilized to analyze the carbon metabolic-pathways network. The analyses revealed (a) variable production of major metabolites ($H_2$, ethanol, acetate, lactate, $CO_2$, and cell mass) depending on initial glucose levels; (b) influence of NADH regeneration on the production of acetate, lactate, and ethanol; and (c) influence of the molar production of ATP on the production of biomass. The results reported in this paper suggest how the carbon metabolic pathway(s) should be designed for optimal Hz production, especially at high glucose concentrations, such as by blocking the carbon flux via lactate dehydrogenase from the pyruvate node.

• KSBB Journal
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• 제29권3호
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• pp.165-178
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• 2014

Statistical medium optimization has been carried out for the production of succinic acid in anaerobic fermentations of Actinobacillus succinogenes. Succinic acid utilized as a precursor of many industrially important chemicals is a fourcarbon dicarboxylic acid, biosynthesized as one of the fermentation products of anaerobic metabolism by A. succinogenes. Through OFAT (one factor at a time) experiments, corn steep liquor (CSL), a very cheap agricultural byproduct, was found to have significant effects on enhanced production of succinic acid, when supplemented along with yeast extract. Hence, using these factors including glucose as a carbon/energy source, interactive effects were investigated through $2^n$ full factorial design (FFD) experiments, showing that the concentration of each component (i.e., glucose, yeast extract and CSL) should be higher. Further statistical experiments were conducted along the steepest ascent path, followed by response surface method (RSM) in order to find out optimal concentrations of each constituent. Consequently, optimized concentrations of glucose, yeast extract and CSL were observed to be 180 g/L, 15.08 g/L and 20.75 g/L respectively (10 g/L of $NaHCO_3$ and 100 g/L of $MgCO_3$ to be supplemented as bicarbonate suppliers), with the estimated production level of succinic acid to be 92.9 g/L (about 3.5 fold higher productivity as compared to the initial medium). Notably, the RSM-estimated production level was almost similar to the amount of succinic acid (92.9 g/L vs. 89.1 g/L) produced through the actual fermentation process performed using the statistically optimized production medium.

• 한국수소및신에너지학회논문집
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• 제13권4호
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• pp.321-329
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• 2002

Clostridium butyricum, Lactobacillus amylophillus, Lactobacillus amylovorus, Lactobacillus acidophillus, AI-9 produced hydrogen and /or organic acids using glucose, lactose and starch at the anaerobic culture conditions. Cl. butyricum NCIB 9576 evolved 1,700 ml H2/L-culture broth and accumulated butyric acid, acetic acid, propionic acid and ethanol in its culture broth when lactose was used as a carbon source during 24 hrs of fermentation. L. amylovorus ATCC 33620 accumulated lactic and acetic acids and some reducing sugars when starch was used as a carbon source without hydrogen production. Instead of starch as a carbon source, L. amylovorus ATCC 33620 produced lactic acid from algal biomass during fermentation and the acid-heat or freeze-thaw pretreatment of algal biomass accelerate the lactic acid fermentation.

• KSBB Journal
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• 제21권6호
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• pp.451-455
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• 2006

본 실험실에서는 음식폐기물을 효율적으로 소화처리하기위해 Pilot 규모 (10톤)의 3단계 메탄 발효 공정을 개발하여 운전하고 있다. 3단계 메탄발효시스템은 반혐기성 가수분해조, 혐기성 산발효조, 혐기성 메탄발효조로 구성되어 있으며, 이 가운데 두 번째 공정인 혐기성 산발효조로부터 알코올발효능이 우수한 균주 KA4를 분리하였다. 세포의 형태는 타원형 모양이며, 크기는 $5.5-6.5{\times}3.5-4.5\;{\mu}m$ 이었고, 26S rDNA D1/D2 rDNA sequence를 분석한 결과를 바탕으로 Saccharomyces cerevisiae KA4로 명명하였다. 이 균주를 YM 배지에서 배양하였을 경우 $30-35^{\circ}C$에서 최대 생장을 보였으며, 배지내의 초기 에탄올 농도가 5% (v/v)까지는 생장에 영향을 받지 않았으나 그 이상에서는 생장에 저해를 받았고 7% 이상에서는 생장하지 못하였다. 한편 초기 50% (w/v)까지의 당 농도에서는 생장이 가능하였으나 잔류 당 농도를 고려할 때 에탄올 발효를 위한 최적 당 농도는 10%이었다. 이 농도의 당을 이용하여 초기 pH4에서 10까지의 넓은 범위에서 에탄올 발효가 가능하였으며 최적 pH는 6이었다 이 때 에탄올 생산량은 7.4%이었으며, 에탄올 생산수율은 2.87 mol EtOH/mol glucose이었다.

• KSBB Journal
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• 제7권2호
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• pp.126-131
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• 1992

P. shermanii IFO 1239을 이용 vitamin $B_{12}$를 생산할 때 산소의 영향은 매우 큼 것으로 나타났는데 배양 전반부는 혐기적 조건이, 후반부는 호기적 조건이 vitamin $B_{12}$ 생성증진에 효과적임이 밝혀졌다. 본 연구를 통하여 배양 48시간 동안은 복합배지상에서 혐기적으로 배양하다가 48시간 이후부터는 합성배지로 바꾸고 호기적 조건으로 배양해주는 방법이 vitamin $B_{12}$ 의 생성을 증가시켰다. 이런 배양방법을 이용하여 탄소원과 유기산이 vitamin $B_{12}$ 생성에 미치는 영향을 알아본 결과 glucose, fructose, lactose가 좋은 탄소원이었으며 유기산 중에서는 succinate 와 malate가 각각 34.9%와 18.7%의 vitamin $B_{12}$생성을 촉진시켰다.

• 한국수소및신에너지학회논문집
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• 제16권4호
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• pp.315-323
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• 2005

The integrated or the two-stage (dark anaerobic and photosynthetic) fermentation processes were compared for the hydrogen production using purple non-sulfur photosynthetic bacteria, Rhodopseudomonas palustris P4. Cell growth, pH changes and organic acids and bacteriochlorophyll contents were monitored during the processes. Culture broth of Rps. palustris P4 exhibited dark-red during the photosynthetic culture condition, while yellow under the anaerobic condition without light. Rps. palustris P4 grown at the photosynthetic condition evolved 0.38 and 1.33 ml $H_2$/mg-dcw during the dark and the light fermentation, respectively, which were totally 1.71 ml $H_2$/mg-dcw at the two-stage fermentation. The rate of hydrogen production using Rps. palustris P4 grown under the dark anaerobic condition was 2.76 ml $H_2$/mg-dcw which consisted of 0.46 and 2.30 ml $H_2$/mg-dcw from the dark and the photosynthetic fermentation processes, respectively. Rps. palustris P4 grown under dark anaerobic conditions produced $H_2$ 1.6 times higher than that of grown under the photosynthetic condition. However, total fermentation period of the former was 1.5 times slower than that of the latter, because the induced time of hydrogen production during the photosynthetic fermentation was 96 and 24 hours when the seed culture was the dark anaerobic and photosynthetic, respectively. The integrated fermentation process by Rps. palustris P4 produced 0.52 ml $H_2$/mg-dcw(1.01 mol $H_2$/mol glucose), which was 20% of the two-stage fermentation.