• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.504-511
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• 2016

Lactulose, a synthetic disaccharide, has received increasing interest because of its role as a prebiotic that can increase the proliferation of Bifidobacterium and Lactobacillus spp. and enhance the absorption of calcium and magnesium. While the industrial production of lactulose is still mainly achieved by the chemical isomerization of lactose in alkaline media, this process has drawbacks including the need to remove catalysts and by-products, as well as high energy requirements. Recently, the use of cellobiose 2-epimerase (CE) has been considered an interesting alternative for industrial lactulose production. In this study, to develop a process for enzymatic lactulose production using CE, we screened improved mutant enzymes ($CS-H^RC^E$) from a library generated by an error-prone PCR technique. The thermostability of one mutant was enhanced, conferring stability up to $75^{\circ}C$, and its lactulose conversion yield was increased by 1.3-fold compared with that of wild-type CE. Using a recombinant Escherichia coli strain harboring a CS35 $H^RC^E$-expressing plasmid, we prepared cell beads immobilized on a Ca-alginate substrate and optimized their reaction conditions. In a batch reaction with 200 g/l lactose solution and the immobilized cell beads, lactose was converted into lactulose with a conversion yield of 43% in 2 h. In a repeated 38-plex batch reaction, the immobilized cell beads were relatively stable, and 80% of the original enzyme activity was retained after 4 cycles. In conclusion, we developed a reasonable method for lactulose production by immobilizing cells expressing thermostable CE. Further development is required to apply this approach at an industrial scale.

• KIPS Transactions on Software and Data Engineering
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• v.11 no.5
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• pp.189-196
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• 2022

Skyline query is a scheme for exploring objects that are suitable for user preferences based on multiple attributes of objects. Existing skyline queries return search results as batch processing, but the need for real-time search results has increased with the advent of interactive apps or mobile environments. Online algorithm for Skyline improves the return speed of objects to explore preferred objects in real time. However, the object navigation process requires unnecessary navigation time due to repeated comparative operations. This paper proposes a Pre-processing Online Algorithm for Skyline Query (POA) to eliminate unnecessary search time in Online Algorithm exploration techniques and provide the results of skyline queries in real time. Proposed techniques use the concept of range-limiting to existing Online Algorithm to perform pretreatment and then eliminate repetitive rediscovering regions first. POAs showed improvement in standard distributions, bias distributions, positive correlations, and negative correlations of discrete data sets compared to Online Algorithm. The POAs used in this paper improve navigation performance by minimizing comparison targets for Online Algorithm, which will be a new criterion for rapid service to users in the face of increasing use of mobile devices.

• Korean Journal of Microbiology
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• v.21 no.2
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• pp.51-60
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• 1983

Cultural characteristics of Strptomyces griseolus isolated from the soil were investigated. This strain was disclosed to utilize D-xylose, and D-glactose in preference order as a carbon source with the formation of glucose isomerase. The addition of sweet potato starch also proved effective promoting the total enzyme activity measured at 29% higher than the control. Corn cob, one of waste agricultural resources, was hydrolyzed in 2~3% $H_2SO_4$ solution at $100^{\circ}C$, 3~5 hours to produce a xylose syrup which gave rise to the recovery of 19.9% in a batch system and 28.2% in a repeated system. By the addition of both 2% of xylose syrup(Be'28) prepared by and us 65% of corn steep liquor (total nitrogen 1.2%), enzyme induction was maximized. The enzyme activity was stimulated by the xylose and the cell growth by the C.S.L. Also, remarkable increase of enzyme activity was noticed by the addition of protein acid hydrolysate 86.2% higher than the control. $QO_2$ of the biomass cultured in 30L capacity jarfermentor recorded low oxygen requirement of 251.2 1/hr. Maximum activity of glucose isomerase was observed noted at the 9th hour after inoculation which is 2 hours faster than the stationery was observed noted at the 9th hour after inoculation which is 2 hours faster than the stationery phase of the biomass growth. Glucose isomerase from the strain was activated by adding the $Co^{++}\;and\;Mg^{++}$ with optimum temperature of $73^{\circ}C$ and pH of 7.2. Conversion ratio of 60% glucose to frutose was 42.5% after 70 hours reaction.

• KSBB Journal
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• v.7 no.2
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• pp.139-143
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• 1992

One of the objectives of this work is to obtain information relevant to the industrial production of alcohol from sugar. The fermentation of alcohol by a strain of saccharomycess cerevisiae ATCC 24858 was studied In a continuous single-stage process with recycle of the cells via tangential flow microfiltration membranes. The experimental results reported in this study pertain to continuous cultures with total cell-recycle by varying the dilution rate (D=0.3, 0.5, and 0.7 $hr^{-1}$) and glucose concentration (50, 100, 150, and 200g/l sugar solution). Productivity using a repeated cell recycle system was found extremely high, 1.e., over 10 to 29 times higher than that of a smile batch system. When a sugar concentration of 200g/1 at dilution rate, 0.7 hr-1 was used, 83.9g/l ethanol was formed with an ethanol yield of 0.42(82% of theoretical) based on sugars utilized.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.110-116
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• 2010

Lactic acid (LA) fermentation by Lactobacillus salivarius ATCC 11741 immobilized on loofa sponge (LS) was evaluated. To increase the surface area of LS for cell immobilization, $H_2O_2$ and chitosan were introduced as surface modifying reagents. Four chitosans of different molecular weights were separately coated on LS. All experiments were conducted in shaking flask mode at 100 rpm rotating speed and $37^{\circ}C$ with 5% $CaCO_3$ as a pH regulating agent. The effects of initial glucose concentration were investigated in the range of 20-100 g/l on LA fermentation by free cells. The results indicate that the maximum concentration of LA was produced with 50 g/l glucose concentration. The immobilized cell system produced 1.5 times higher concentration than free cells for 24 h of fermentation. Moreover, immobilized cells can shorten the fermentation time by 2-fold compared with free cells at the same level of LA concentration. At 1% (w/v) chitosan in 2% (v/v) acetic acid, the Yp/s and productivities of various molecular weights of chitosans were insignificantly different. Repeated batch fermentations showed 5 effective recycles with Yp/s and productivity in the range of 0.55-0.85 and 0.90-1.20 g/l.h, respectively. It is evident that immobilization of L. salivarius onto LS permits reuse of the system under these fermentation conditions. Scanning electron micrographs indicated that there were more intact cells on the chitosan-treated LS than on the untreated LS, thus confirming the effectiveness of the LS-chitosan combination when being utilized as a promising immobilization carrier for LA fermentation.

• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1227-1234
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• 2020

In this study, yeast cell immobilization was carried out in a packed bed reactor (PBR) to investigate the effects of the volumetric capacity of carriers as well as the different fermentation modes on fuel ethanol production. An optimal volumetric capacity of 10 g/l was found to obtain a high cell concentration. The productivity of immobilized cell fermentation was 16% higher than that of suspended-cell fermentation in batch and it reached a higher value of 4.28 g/l/h in repeated batches. Additionally, using this method, the ethanol yield (95.88%) was found to be higher than that of other tested methods due to low concentrations of residual sugars and free cells. Continuous ethanol production using four bioreactors showed a higher productivity (9.57 g/l/h) and yield (96.96%) with an ethanol concentration of 104.65 g/l obtained from 219.42 g/l of initial total sugar at a dilution rate of 0.092 h^-1. Furthermore, we reversed the substrate-feed flow directions in the in-series bioreactors to keep the cells at their highest activity and to extend the length of continuous fermentation. Our study demonstrates an effective method of ethanol production with a new immobilized approach, and that by switching the flow directions, traditional continuous fermentation can be greatly improved, which could have practical and broad implications in industrial applications.

• Journal of Microbiology and Biotechnology
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• v.23 no.10
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• pp.1434-1444
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• 2013

We established a two-step production process using immobilized S. cerevisiae and P. stipitis yeast to produce ethanol from seaweed (U. pertusa Kjellman) hydrolysate. The process was designed to completely consume both glucose and xylose. In particular, the yeasts were immobilized using DEAE-corncob and DEAE-cotton, respectively. The first step of the process included a continuous column reactor using immobilized S. cerevisiae, and the second step included a repeated-batch reactor using immobilized P. stipitis. It was verified that the glucose and xylose in 20 L of medium containing the U. pertusa Kjellman hydrolysate was converted completely to about 5.0 g/l ethanol through the two-step process, in which the overall ethanol yield from total reducing sugar was 0.37 and the volumetric ethanol productivity was 0.126 g/l/h. The volumetric ethanol productivity of the two-step process was about 2.7 times greater than that when P. stipitis was used alone for ethanol production from U. pertusa Kjellman hydrolysate. In addition, the overall ethanol yield from glucose and xylose was superior to that when P. stipitis was used alone for ethanol production. This two-step process will not only contribute to the development of an integrated process for ethanol production from glucose-and xylose-containing biomass hydrolysates, but could also be used as an alternative method for ethanol production.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.257-260
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• 2002

Dissolved oxygen level of cell culture media has a critical effect on cellular metabolism, which governs specific productivity of recombinant proteins and mammalian cell growth However, in the cores of cell aggregates or cell-immobilized beads, oxygen level frequently goes below a critical level. Mammalian cells have a number of genes induced in the lower level of oxygen, and the genes contain a common cis-acting element (-RCGTG-), hypoxia response element (HRE). By binding of hypoxia inducible factor-l (HIF-I) to the HRE, promoters of hypoxia inducible genes are activated, which is a survival mechanism. In this work, to develop a CHO cell capable of producing recombinant proteins in immobilization and high density cell culture efficiently, mammalian expression vectors containing human tissue-type plasminogen activator (t-PA) gene controlled by HRE were constructed and stably transfected into the CHO cells. In $Ba^{2+}$ -alginate immobilization culture, CHO/pCl/dhfr/2HRE-t-PA cells produced 2 folds higher recombinant t-PA activity than CHO/pCl/dhfrlt-PA cells without $CoCl_2$ treatment. Furthermore, in repeated fed batch culture, productivity of t-PA in immobilized CHO/pCI/dhfr/2HRE-t-PA cells was 121 ng/ml/day, total production of 0.968 mg/day at 11 days culture while CHO/pCIIdhfrlt-PA cells was 22.8 ng/ml/day. All these results indicate that HRE is very useful for the enhancement of protein productivity in mammalian cell cultures.

• Journal of Korean Society on Water Environment
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• v.36 no.4
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• pp.322-331
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• 2020

By introducing chemically enhanced primary treatment (CEPT) in the first stage of sewage treatment, organic matter in sewage can be effectively recovered. Because CEPT sludge contains a high biodegradable organic matter in volatile solids (VS), it is feasible to convert the collected CEPT sludge into energy through anaerobic digestion. This study examined the properties and biochemical methane potential (BMP) of the CEPT sludge obtained from a sewage treatment plant located in an ocean area. The CEPT sludge contains a VS content of 37,597 mg/L, which is higher than that of excessive sludge (ES), i.e., 33,352 mg-VS/L. In the methane generation reaction, the lag period was as short as 1 to 2 days. The BMP for the CEPT sludge was 0.57 ㎥-CH₄/kg-VS_removed which is better than that of ES, i.e., 0.36 ㎥-CH₄/kg-VS_removed. Unfortunately, the CEPT sludge showed a high salinity as 0.56~0.75% probably due to the saline sewage. Due to the salinity, repeated BMP testing in a sequencing batch reactor showed significantly low methane production rates and BMPs. Also, the ES showed a strongly reduced BMP when the salinity was adjusted from 0.20 to 0.70% by NaCl. The ES mixture with higher CEPT content showed a better BMP, which is suitable for co-digestion. Besides, anaerobic digestion for 100% CEPT sludge can be a considerable option instead of co-digestion.

• KSBB Journal
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• v.32 no.1
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• pp.71-82
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• 2017

To verify anti-diabetic effect of oligopeptide with His-Pro repeats (mHP peptide), the oligopeptide was first secreted and optimized using the secretion vector, pRBAS with alkaline protease gene promoter and the signal sequence in Bacillus subtilis and directly the anti-diabetic effect of the mHP peptide was investigated in insulinoma cell, RINm5F cell line. The oligopeptide gene was obtained by annealing oligonucleotides with repeated His-Pro sequence and finally was constructed as 18 dipeptides (108 bp and 4.0 kDa) coding gene, named oligopeptide with His-Pro repeats (mHP peptide) to make cyclo(His-Pro) known to be anti-diabetic effects. The region encoding the oligopeptide gene was subcloned into the pRBAS secretion vector (E.coli-Bacillus shuttle vector) after PCR amplification using the designed primers including initiation and termination codons and His tag, named pRBAS-mHP (6.56 kb). To optimize secretion of the oligopeptide, various culture conditions were investigated in Bacillus subtilis LKS. As a result, the secreted oligopeptide was maximally measured (approximately $59.6{\mu}g/mL$) in 3 L batch culture and the highest secretion was achieved at $30^{\circ}C$, PY medium, and carbon sources (particularly barley and glycerol). In the RINm5F cells treated with 2 mM STZ, the oligopeptide treatment (0.1 mg/mL) restored the cell viability (10%) and reduced the nitric oxide (NO) generation (35%) and DNA fragmentation (90%). And also, insulin secretion level was increased to 17% higher than in STZ-treated RINm5F cells. These results suggest that the oligopeptide with His-Pro repeats could be a candidate material for anti-diabetic agent against STZ-induced diabetes.