• Journal of Microbiology and Biotechnology
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• v.24 no.4
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• pp.431-439
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• 2014

We report the construction of two Bacillus subtilis expression vectors, pBNS1/pBNS2. Both vectors are based on the strong promoter P43 and the ampicillin resistance gene expression cassette. Additionally, a fragment with the Shine-Dalgarno sequence and a multiple cloning site (BamHI, SalI, SacI, XhoI, PstI, SphI) were inserted. The coding region for the amyQ (encoding an amylase) signal peptide was fused to the promoter P43 of pBNS1 to construct the secreted expression vector pBNS2. The applicability of vectors was tested by first generating the expression vectors pBNS1-GFP/pBNS2-GFP and then detecting for green fluorescent protein gene expression. Next, the mannanase gene from B. pumilus Nsic-2 was fused to vector pBNS2 and we measured the mannanase activity in the supernatant. The mannanase total enzyme activity was 8.65 U/ml, which was 6 times higher than that of the parent strain. Our work provides a feasible way to achieve an effective transformation system for gene expression in B. subtilis and is the first report to achieve B. pumilus mannanase secretory expression in B. subtilis.

• Membrane and Water Treatment
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• v.14 no.1
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• pp.35-45
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• 2023

Biofilms significantly affect the performance of wastewater treatment processes in which biodegradability of numerous microorganisms are actively involved, and various technologies have been applied to secure microbial biofilms. Understanding changes in biofilm characteristics by regulating expression of signaling molecules is important to control and regulate biofilms in membrane bioreactor, i.e., biofouling. This study investigated effects of addition of acyl-homoserine lactones (AHL) as a controllable factor for the microbial signaling system on biofilm formation of Pseudomonas aeruginosa PAO1 and multiple strains in membrane bioreactor. The addition of three AHL, i.e., C4-, C6-, and C8-HSL, at a concentration of 200 ㎍/L, enhanced the formation of the PAO1 biofilm and the degree of increases in the biofilm formation of PAO1 were 70.2%, 76.6%, and 72.9%, respectively. The improvement of biofilm formation of individual strains by C4-HSL was an average of 68%, and the microbial consortia increased by approximately 52.1% in the presence of 200 ㎍/L C4-HSL. CLSM images showed that more bacterial cells were present on the membrane surface after the AHL application. In the COMSTAT results, biomass and thickness were increased up to 2.2 times (PAO1) and 1.6 times (multi-strains) by C4-HSL. This study clearly showed that biofilm formation was increased by the application of AHL to individual strain groups, including PAO1 and microbial consortia, and significant increases were observed when 50 or 100 ㎍/L AHL was administered. This suggests that AHL application can improve the biofilm formation of microorganisms, which could yield an enhancement in efficiency of biofilm control, such as in various biofilm reactors including membrane bioreactor and bioflocculent systems in water/wastewater treatment processes.

• Korean Journal of Food Science and Technology
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• v.35 no.1
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• pp.103-110
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• 2003

Stable production of fermented kanjang containing 1.8% (v/v) ethanol was obtained within four days using traditional kanjang containing 4% added glucose in packed-bed bioreactor systems filled with immobilized Zygosaccharomyces rouxii and Candida versatilis on porous alumina ceramic bead carrier at $28{\pm}0.5^{\circ}C$ and aeration rate of 0.05 vvm. Specific rates of alcohol production for Z. rouxii and C. versatilis were 0.0033 and 0.0031/day, respectively, and those of glucose consumption were both -0.0087/day in the batch type of alcoholic fermentation. In semi-continuous alcoholic fermentation at a dilution rate of 0.25/day, specific rates of alcohol production for Z. rouxii and C. versatilis were 0.0045 and 0.0029/day, and those of glucose consumption were -0.01 and -0.008/day, respectively, using identical bioreactor system. Similar specific rates of alcohol production were observed both in the batch or semi-continuous process and in the continuous one at the dilution rate of 0.25/day. Sensory characteristics of all alcoholic-fermented kanjang by Z. rouxii, C. versatilis, and a mixture of both yeasts (2:1, w/w) were shown to be significantly superior to those of home-made kanjang as revealed through organoleptic evaluation tests (p<0.05).

• Reproductive and Developmental Biology
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• v.40 no.1
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• pp.7-13
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• 2016

Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat ${\beta}$-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

• Journal of Microbiology and Biotechnology
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• v.28 no.1
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• pp.145-156
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• 2018

Most eukaryotic peroxiredoxins (Prxs) are readily inactivated by a high concentration of hydrogen peroxide ($H_2O_2$) during catalysis owing to their "GGLG" and "YF" motifs. However, such oxidative stress sensitive motifs were not found in the previously identified filamentous fungal Prxs. Additionally, the information on filamentous fungal Prxs is limited and fragmentary. Herein, we cloned and gained insight into Aspergillus nidulans Prx (An.PrxA) in the aspects of protein properties, catalysis characteristics, and especially $H_2O_2$ tolerability. Our results indicated that An.PrxA belongs to the newly defined family of typical 2-Cys Prxs with a marked characteristic that the "resolving" cysteine ($C_R$) is invertedly located preceding the "peroxidatic" cysteine ($C_P$) in amino acid sequences. The inverted arrangement of $C_R$ and $C_P$ can only be found among some yeast, bacterial, and filamentous fungal deduced Prxs. The most surprising characteristic of An.PrxA is its extraordinary ability to resist inactivation by extremely high concentrations of $H_2O_2$, even that approaching 600 mM. By screening the $H_2O_2$-inactivation effects on the components of Prx systems, including Trx, Trx reductase (TrxR), and Prx, we ultimately determined that it is the robust filamentous fungal TrxR rather than Trx and Prx that is responsible for the extreme $H_2O_2$ tolerence of the An.PrxA system. This is the first investigation on the effect of the electron donor partner in the $H_2O_2$ tolerability of the Prx system.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.8
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• pp.886-891
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• 2005

In this study, a novel bioreactor system using a submerged hollow fiber membrane module (so called hollow fiber membrane bioreactor, HFMB) was applied to investigate feasibility and biodegradation capacity of the system for the treatment of gaseous toluene. First an abiotic test was conducted to determine the mass transfer coefficient, showing the value was similar to that obtained from a diffuser system using fine bubbles. Second, in the presence of toluene-degrading microorganisms, the HFMB was operated at different inlet toluene loading rates of 50, 100, $500\;g/m^3/hr$, and overall removal efficiencies were maintained in the range of $70{\sim}80%$. In addition, elimination capacities(EC) were increased up to $800\;g/m^3/hr$, which was substantially higher than maximum ECs for toluene reported in the biofiltration literature. Consequently, the HFMB was considered as an alternative method over other conventional VOC-treating technologies.

• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.2
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• pp.322-327
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• 1994

For the improvement of L-lysine productivity, development of the continuous fermentation system by a bioreactor assembly was attempted. Primarily, optimal conditions on the whole cell immobilization of Corynebacterium glutamicum ATCC21514 were studied and 76.2% of immobilization ratio was obtained when the cells were entrapped with 4% k-carrageenan showing 4.0kg gel strength. A bioreactor system was set up using the immobilized cells was applied for the continuous production of L-lysine. The results obtained under the optimum conditions were compared with those of the batchwise fermentation. Experimental results obtained from 14 day continuous fermentation showed 36.7% of sugar conversion to L-lysine while the productivity of L-lysine was disclosed as 4.96mg/ml mg-dry cell weight /hr which is 2.5times and 4.1 times higher than those of the batchwise fermentation by the intact cells and by the immobilized cells, respectively.

• Environmental Engineering Research
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• v.19 no.4
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• pp.387-393
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• 2014

The purpose of this study is to evaluate integrated anaerobic hydrogen fermentation and membrane bioreactor (MBR) for on-site domestic wastewater treatment and resource recovery. A synthetic wastewater (COD 17,000 mg/L) was used as artificial brown water which will be discharged from urine diversion toilet and fed into a continuous stirred tank reactor (CSTR) type anaerobic reactor with inclined plate. The effluent of anaerobic reactor mixed with real household grey water (COD 700 mg/L) was further treated by MBR for reuse. An optimum condition maintained in anaerobic reactor was HRT of 8 hrs, pH 5.5, SRT of 5 days and temperature of $37^{\circ}C$. COD removal of 98% was achieved from the overall system. Total gas production rate and hydrogen content was 4.6 L/day and 52.4% respectively. COD mass balance described the COD distribution in the system via reactor byproducts and effluent COD concentration. The results of this study asserts that, anaerobic hydrogen fermentation combined with MBR is a potent system in stabilizing waste strength and clean hydrogen recovery which could be implemented for onsite domestic wastewater treatment and reuse.

• Membrane and Water Treatment
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• v.9 no.5
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• pp.343-351
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• 2018

As membrane fouling is based on various factors, it is a complex phenomenon that is hard to estimate. This study investigated membrane fouling in a thermophilic jet loop membrane bioreactor (JLMBR). With this purpose, four different empirical membrane fouling models with different sludge retention times were applied on the flow data obtained in the system. As a result of the model implementation, it was found for all sludge retention times that, standard blocking is effective in the first 1.5 hours of filtration, while cake filtration was dominant in the remaining duration. Additionally, it was observed that as the sludge retention time increases, membrane fouling rate decreases.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2003.09a
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• pp.326-329
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• 2003

When an electrokinetic process is applied to a HOC-contaminated soil, hybrid types combined with soil flushing, chemical oxidation, and bioremediation are generally used. Especially when the electrokinetic process is combined with bioremediation, the hybrid technology can solve several limits of bioremediation such as low microbial mobility, low soil temperature, and shortage of nutrients in subsurface circumstance. Because microbial surface is charged negatively, the microorganism moves from cathode to anode under electrical field. In this study, mixed culture mainly-consisted by Pseudomonas sp. was applied to remediate pentadecane-contaminated kaolinite with particle size less than 300${\mu}{\textrm}{m}$. This remediation system was named ‘electrokinetic bioaugmentation’ and consisted of model aquifer, electrode reservoirs, bioreactor, power supply, and pump. The mixed culture above 0.5 of optical density in bioreactor was supplied to two reservoirs and penetrated soil when the electric current was applied. To enhance the removal efficiency, the optimal medium composition, electric current, and voltage were investigated.