• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1928-1935
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• 2015

Microbial growth kinetics is often used to optimize environmental processes owing to its relation to the breakdown of substrate (contaminants). However, the quantification of bacterial populations in the environment is difficult owing to the challenges of monitoring a specific bacterial population within a diverse microbial community. Conventional methods are unable to detect and quantify the growth of individual strains separately in the mixed culture reactor. This work describes a novel quantitative PCR (qPCR)-based genomic approach to quantify each species in mixed culture and interpret its growth kinetics in the mixed system. Batch experiments were performed for both single and dual cultures of Pseudomonas putida and Escherichia coli K12 to obtain Monod kinetic parameters (μ_max and K_s). The growth curves and kinetics obtained by conventional methods (i.e., dry weight measurement and absorbance reading) were compared with that obtained by qPCR assay. We anticipate that the adoption of this qPCR-based genomic assay can contribute significantly to traditional microbial kinetics, modeling practice, and the operation of bioreactors, where handling of complex mixed cultures is required.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1252-1256
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• 2008

Cell recycled culture of succinic acid-producing Anaerobiospirillum succiniciproducens was anaerobically carried out using an internal membrane filter module in order to examine the physiological response of A. succiniciproducens to a high-cell-density environment. The optimal growth of A. succiniciproducens and its enhanced succinic acid productivity were observed under $CO_2$-rich conditions, established by adding $NaHCO_3$ and $Na_2CO_3$, in the cell recycled system. A. succiniciproducens grew up to 6.50 g-DCW/l, the highest cell concentration obtained so far, in cell recycled cultures. The cells did not change their morphology, which is known to be easily changed in unfavorable or stress environments. The maximum productivity of succinic acid was about 3.3 g/l/h, which is 3.3 times higher than those obtained in batch cultures. These results can serve as a guide for designing highly efficient cell recycled systems for succinic acid at a commercial level.

• Animal cells and systems
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• v.3 no.2
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• pp.153-159
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• 1999

In an artificial pH-gradient batch culture system, the effects of acidification on the species composition of a heterotrophic bacterial community were analyzed. As a result of this study, it was found that total bacteria numbers were not affected by acidification and that the population of hetero-trophic bacteria decreased as pH became lower. The heterotrophic bacteria isolated from the entire pH gradient were 12 genera and 22 species. Among them, 64% were gram negative and 36% were gram positive bacteria. As pH decreased, the distribution rate of gram negative bacteria increased while that of gram positive bacteria decreased. The diversity of genera decreased from 13 to 5 as pH decreased from 7 to 3. The G+C content of all of the 202 isolated strains varied from 22.8 to 77.0%, and increased in interspecies of same genus as pH decreased. As a result of clustering analysis, the diversity index of species ranged from 1.13 to 2.37, and it had lower indices as pH decreased. In order to evaluate the diversity of numbers of sample of different size, a rarefaction method was used to analyze the expected number of species appearance according to pH. The statistical significance of species diversity was verified by the fact that the number decreased at lower pH.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.333-338
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• 1987

In order to establish the biological treatment system which can be used for treatment of waste aster from acetaldehyde plant, it was investigated optimum nutrient requirements and growth conditions by mixed culture of Micrococcus roseus AW-6, Micrococcus luteus AW-22, Microbacterium lacticum AW-38 and Microbacterium laevaniformans AW-41 as well as the effect of coagulants and neutralization reagents. Also, it was carried out the continuous culture as well as batch culture to treat the waste water by mixed culture of these strains. The COD removal rate was reached to maximum state for 96hrs culture at pH7.0 and $30^{\circ}C$ NaOH as the neutralization reagents was the most effective, but the coagulants had no effect on the COD remonal rate and the optimum dilution times for treatment were 10 fold. The COD removal rate was also increased by supplimenting 200 ppm $NH_{2}NO_{3}$, 50 ppm $KH_{2}PO_{4}$, 15 ppm $CaCl_{2}$ and 1 ppm $MgSO_{4} \cdot 7H_{2}O $ as additional nutrients. The removal rate coefficient $K_{1}$ on the batch culture was $4.5\times 10^{-6}$, and the detention time for BOD removal rate of 85% was approximately 45hrs. The COD of waste water was reduced to 15% of its initial value by the continuous culture. The COD and BOD of the effluents were to be about 60 ppm and 40 ppm, respectively, and final pH was 7.0.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.400-406
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• 2012

To improve the hydrogen yield from biological fermentation of organic wastewater, a co-culture system of dark- and photo-fermentation bacteria was investigated. In a pure-culture system of the dark-fermentation bacterium Clostridium butyricum, a pH of 6.25 was found to be optimal, resulting in a hydrogen production rate of 18.7 ml-$H_2/l/h$. On the other hand, the photosynthetic bacterium Rhodobacter sphaeroides could produce the most hydrogen at 1.81mol-$H_2/mol$-glucose at pH 7.0. The maximum specific growth rate of R. sphaeroides was determined to be 2.93 $h^{-1}$ when acetic acid was used as the carbon source, a result that was significantly higher than that obtained using either glucose or a mixture of volatile fatty acids (VFAs). Acetic acid best supported R. sphaeroides cell growth but not hydrogen production. In the co-culture system with glucose, hydrogen could be steadily produced without any lag phase. There were distinguishable inflection points in a plot of accumulated hydrogen over time, resulting from the dynamic production or consumption of VFAs by the interaction between the dark- and photo-fermentation bacteria. Lastly, the hydrogen production rate of a repeated fed-batch run was 15.9 ml-$H_2/l/h$, which was achievable in a sustainable manner.

• Journal of Microbiology and Biotechnology
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• v.19 no.5
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• pp.468-473
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• 2009

The antibiotic meroparamycin was produced in the free culture system of Streptomyces sp. strain MAR01. Five solid substrates (rice, wheat bran, Quaker, bread, and ground corn) were screened for their ability to support meroparamycin production in solid-state fermentation. In batch culture, wheat bran recorded the highest antibacterial activity with the lowest residual substrate values. The highest residual substrate values were recorded for both ground corn and Quaker. On the other hand, no antibacterial activity was detected for rice as a solid substrate. The use of the original strength of starch-nitrate medium in the solid-state fermentation gave a lower antibacterial activity compared with the free culture system. Doubling the strength of this medium resulted in the increase in the activity to be equivalent to the free culture. The initial pH (7.0) of the culture medium and 2 ml of spore suspension (1 ml contains $5{\times}10^{9}spores/ml$) were the optima for antibiotic production. The water was the best eluent for the extraction of the antibiotic from the solid-state culture. Ten min was enough time to extract the antibiotic using a mixer, whereas, 60 min was required when shaking was applied. Semicontinuous production of meroparamycin using a percolation method demonstrated a more or less constant antibacterial activity over 4 runs ($450-480{\mu}g/ml$). The semicontinuous production of the antibiotic was monitored in a fixed-bed bioreactor and the maximum activity was attained after the fourth run ($510{\mu}g/ml$) and the overall process continued for 85 days.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.135-141
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• 2004

Human granulocyte colony-stimulating factor (hG-CSF) is a hematopoiesis agent that principally affects the differentiation of neutrophils in the bone marrow. At present, recombinant hG-CSF is used successfully in the treatment of chemotheraphy-induced neutropenia and its indication has been expanded to bone marrow transplantation and aplastic anemia. In this study, we have constructed rhG-CSF secretion plasmid pYRC1 in which OmpA signal sequence/hG-CSF gene was expressed under the control of the T7 promoter. rhG-CSF produced in E. coli BL21 (pYRC1) grown at $37{\circ}C$ was found in aggregates. However, 15% of the periplasmic protein was soluble rhG-CSF when the E. coli BL21 (pYRC1) was cultured at $25^{\circ}C$ for 7 h in the modified MBL medium containing 10 g/$\ell$ glucose with 10 $\mu$M IPTG induction. The production of soluble rhG-CSF in E. coli BL21 (pYRC1) using fed batch culture was also studied. In the fed batch culture system, the final yield of rhG-CSF produced from E. coli BL21 (pYRC1) was increased from 4.4 mg/$\ell$to 24 mg/$\ell$by controlling the specific growth rate from $0.43 h^{-1}$ to $0.14 h^{-1}$, and optimizing the time of induction.

• Journal of Korean Society on Water Environment
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• v.21 no.2
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• pp.118-124
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• 2005

The oxygen solubilized device(O.S.D) and standardized microorganism culture system is more efficient than physical and chemical purification techniques in closed water. This study was to determine how the O.S.D and standardized culture system is efficient in purification capacity in closed water based on the lab scale and pilot plant. In the batch test, inducing the quantitative results from pilot plant operation condition, removal efficiency of COD and TN were about 48.3% and 35% respectively, while SS and chlorophyll-a were 94.9% and 68.7%. The pilot plant results showed that suspended solid(SS) and chlorophyll-a removal efficiency were 60% and 59% respectively, due to coagulation characteristics by standardized culture. Total nitrogen(TN) and total phosphorus(TP)showed good effect for the purification of target pond water quality from field data. Additionally, released velocity was determined in control condition of $5.31mgPO{_4}^{3-}{\cdot}m^{-2}{\cdot}day^{-1}$ and $2486.8mgCOD{\cdot}m^{-2}{\cdot}day^{-1}$. Otherwise, phosphate and COD reflux in the aeration and microorganism condition was showed $-9.95mgPO{_4}^{3-}{\cdot}m^{-2}{\cdot}day^{-1}$ and $-397.88mgCOD{\cdot}m^{-2}{\cdot}day^{-1}$. This technology is the most effective not only removal of SS and chlorophyll-a but also control of phosphate and COD release which is very important phenomena in evaluating water quality in closed water like a reservoir and pond.

• KSBB Journal
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• v.11 no.5
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• pp.524-528
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• 1996

Bacterial degradation of 4-chloro-2-methylphenoxyacetic acid (MCPA) was studied in column reactors under conditions approximating a fluidized bed system, with granular activated carbon (GAC) as a support matrix. A mixed bacterial culture of MCPA-degrading bacteria was used as an inoculum to develop a biofilm on GAC. Initially, adsorption of MCPA by GAC and blofilm formation on GAC were examined. MCPA degradation was evaluated with a batch and continuous mode of operation of the GAC fixed-film column reactors. In the batch operations, complete degradation of MCPA was achieved during the incubation period. Partial degradation of MCPA occurred in the continuous operations and MCPA degradation was dependent on the feeding rate of MCPA solution.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.753-759
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• 2002

$\beta$-Cyclodextrin glucanotransferase ($\beta$-CGTase) was overproduced extracellularly using recombinant E. coli by transforming the plasmid pECGT harboring a secretive signal peptide. The $\beta$-CGTase gene of alkalophilic Bacillus firmus var alkalophilus was inserted into the high expression vector pET20b(+) containing a secretive pelB signal peptide, and then transformed into E. coli BL2l(DE3)pLysS. The optimum culture conditions fer the overproduction of $\beta$-CGTase were determined to be TB medium containing 0.5% (w/v) soluble starch at post-induction temperature of $25^{\circ}C$. A significant amount of $\beta$-CGTase, up to 5.83 U/ml, which was nine times higher than that in the parent strain B. firmus var. alkalophilus, was overproduced in the extracellular compartment. A pH-stat fed-batch cultivation of the recombinant E. coli was also performed to achieve the secretive overproduction of $\beta$-CGTase at a high cell density, resulting in production of up to 21.6 U/ml of $\beta$-CGTase.