• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.356-359
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• 2000

A two-stage fed-batch fermentation was carried out to increase xylitol productivity by Candida tropicalis. The first stage for cell growth was performed in the pH-stat and continuous fed-batch modes. The higher cell growth and lower ethanol production obtained in the fed-batch mode where the growth medium was fed when pH of culture broth increased over 5.7. And also the effect of oxygen transfer on xylitol production was investigated by changing agitation speed under 0.5 vvm of aeration. The maximum xylitol productivity and yield were obtained at 500 rpm of agitation.

• Journal of Microbiology and Biotechnology
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• v.19 no.5
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• pp.474-481
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• 2009

A novel microorganism, designated as LG12, was isolated from soil based on its ability to use acrylic acid as the sole carbon source. An electron microscopic analysis of its morphological characteristics and phylogenetic classification by 16S rRNA homology showed that the LG12 strain belongs to Rhodococcus erythropolis. R. erythropolis LG12 was able to metabolize a high concentration of acrylic acid (up to 40 g/l). In addition, R. erythropolis LG12 exhibited the highest acrylic acid-degrading activity among the tested microorganisms, including R. rhodochrous, R. equi, R. rubber, Candida rugosa, and Bacillus cereus. The effect of the culture conditions of R. erythropo/is LG12 on the production of 3-hydroxypropionic acid (3HP) from acrylic acid was also examined. To enhance the production of 3HP, acrylic acid-assimilating activity was induced by adding 1 mM acrylic acid to the culture medium when the cell density reached an $OD_{600}$ of 5. Further cultivation of R. erythropo/is LG 12 with 40 g/l of acrylic acid resulted in the production of 17.5 g/l of 3HP with a molar conversion yield of 44% and productivity of 0.22 g/l/h at $30^{\circ}C$ after 72 h.

• Food Science and Biotechnology
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• v.18 no.4
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• pp.954-958
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• 2009

Effects of controlled- and uncontrolled-pH operations on phenylalanine ammonia lyase (PAL) production by a recombinant Escherichia coli strain were investigated at uncontrolled-pH ($pH_{UC}$) and controlled-pH ($pH_C$) of 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5 in bioreactor systems. The results showed that the recombinant PAL activity was improved significantly by controlled pH strategy. Among the $pH_C$ operations, the highest PAL activities were obtained under $pH_C$ 7.5 strategy where cell mass ($OD_{600\;nm}$) and PAL activity was 1.3 and 1.8 fold higher than those of $pH_{UC}$, respectively. The maximum PAL activity reached 123 U/g. The $pH_C$ 7.5 strategy made recombinant plasmid more stable and therefore allowed easier expression of PAL recombinant plasmid, which increased PAL production. It was indicated that the new approach (controlled-pH strategy) obtained in this work possessed a high potential for the industrial production of PAL, especially in the biosynthesis of L-phenylalanine.

• Journal of Life Science
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• v.24 no.10
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• pp.1046-1054
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• 2014

Beta-1,3-glucanase is widely used in various biotechnological and industrial processes, with over-production required to enable versatile utilization. We examined the overexpression of ${\beta}$-1,3-glucanase (EXGA) from Aspergillus oryzae using ${\delta}$-sequence-mediated integration. We constructed $pRS{\delta}$-exgA and $pRS{\delta}K$-exgA plasmids for integration of the EXGA gene into various chromosomes of Saccharomyces cerevisiae. These plasmids contain the ADH1 promoter for constitutive expression, a signal sequence (exoinulinase signal sequence [INU1 s.s]) for secretory production, and a ${\delta}$-sequence for integration of ${\beta}$-1,3-glucanase. The $pRS{\delta}$-exgA plasmid was transformed into the S. cerevisiae $BY4742{\Delta}exg1$ strain, and ${\beta}$-1.3-glucanase was stably overexpressed and secreted. Another plasmid, $pRS{\delta}K$-exgA, was introduced into the S. cerevisiae $BY4742{\Delta}exg1$ (YKY082) strain, and overexpression of ${\beta}$-1,3-glucanase was examined by inducible integration under geneticin selection. The activity of ${\beta}$-1,3-glucanase increased in accordance with a rise in the geneticin concentration, with 0.8 mg/ml of geneticin suitable for overexpression of ${\beta}$-1,3-glucanase. Subsequently, $pRS{\delta}K$-exgA was repeatedly transformed for sequential ${\delta}$-integration. The activity of ${\beta}$-1,3-glucanase reached about 0.063 unit/ml/$OD_{600}$, 0.095 unit/ml/$OD_{600}$, 0.131 unit/ml/$OD_{600}$ and 0.165 unit/ml/$OD_{600}$ by the first, second, third, and fourth round of integration, respectively. According to the increase in the activity of ${\beta}$-1,3-glucanase by sequential ${\delta}$-integration, the copy number (integration rate) of the EXGA gene also increased in various chromosomes. These results suggest that recombinant ${\beta}$-1,3-glucanase activity can be sequentially increased by repeated ${\delta}$-sequence integration.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.982-986
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• 2009

For the production of ghost bacteria vaccine to prevent the streptococcal disease in aquaculture fish species, a double cassettes vector was constructed and cloned in Escherichia coli DH5${\alpha}$. Ghost bacteria vaccine production from Escherichia coli DH5${\alpha}$/pHCE-InaN-GAPDH-Ghost 37 SDM (SIG) was maximized at a glucose concentration of 1 g/l, agitation of 300 rpm, and aeration of 1 vvm. The maximal efficiency of ghost bacteria formation was obtained at the mid-exponential phase ($OD_{600}=2.0$) with the concentration of 0.77 g/l for SIG. The molecular mass of GAPDH was detected at 67 kDa with the insoluble fraction, by SDS-PAGE and Western blot. The protective efficacy of ghost bacteria vaccine was evaluated by challenge test using olive flounder. The cumulative mortalities of the positive control, formalin-killed cell (FKC) vaccine, and SIG vaccine immunized groups were 91%, 74%, and 57%, respectively. These results suggest that SIG vaccine showed efficacy as a vaccine and had a higher potential to induce protective antibodies than did FKC vaccine.

• Microbiology and Biotechnology Letters
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• v.27 no.4
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• pp.327-332
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• 1999

A highly productive fed-batch fermentation process was developed for the production of L-ornithine by using a new stabilized strain, Breviabcterium ketoglutamicum BK52. Fed-batch cultures with a continuous feeding of the complex medium were conducted on various operating conditions. The optimal concentration of phosphate in the complex medium was 2.1g/L. The optimal feeding rate of L-arginine was 0.028g/L/hr. The optimal feeding point of the complex medium was determined to be at 40 OD of the cell mass. The final L-ornithine concentrations within 64hrs of cultivation in 5 and 50 liter fermenters were 73g/L and 71g/L, respectively. The maximum overall L-ornithine productivity was 1.14g/L/hr which was about 2 times higher than that of the conventional fed-batch culture with intermittent feeding. The overall productivity of the fermentation system is remarkably improved by employing the optimized conditions, and it offers a significant potential for industrial application.

• KSBB Journal
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• v.26 no.3
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• pp.223-228
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• 2011

The effects of pH, glycerol concentration and salt on cell growth and ethanol production using Enterobacter aerogenes KCTC 2190 were evaluated in the anaerobic culture condition. In condition of initial pH 5, cell growth and ethanol production were highest. An initial concentration of 10 g/L of pure glycerol gave the highest cell growth and ethanol production. However, in case of over 15 g/L of pure glycerol, they decreased. The cell growth and ethanol production decreased with the increase of salt concentration. When 10 g/L of crude glycerol was used as the carbon source, the cell growth and ethanol production were $1.32\;OD_{600}$ and 3.95 g/L, respectively, which were about 94.4% and 88.5% compared to those of pure glycerol. These result indicates that the crude glycerol produced in the biodiesel manufacturing process maybe useful as a potential carbon source for ethanol production form Enterobacter aerogenes KCTC 2190.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.2
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• pp.169-174
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• 2017

Immunoglobulin M (IgM) was purified from olive flounder Paralichthys olivaceus sera using mannan-binding protein (MBP) and protein L affinity columns (designated as MBPIgM and ProLIgM, respectively). A monoclonal antibody (MAb) against olive flounder IgM was produced. The MBPIgM and ProLIgM had apparent molecular weights of 77, 73, and 28 kDa in SDS-PAGE. Nine hybridomas secreting MAbs against olive flounder IgM were established: five MAbs for MBPIgM (1, 2, 3, 4, and 5) and four for ProLIgM (6, 7, 8, and 9). Western blotting indicated that seven MAbs recognized heavy (H; MAbs 1, 2, 3, 4, 5, 6, and 7) chains and one recognized light (L; MAb 9) chains of IgM, while MAb 8 did not recognize IgM. The results of enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the nine MAbs revealed that the optical density (OD) values of sera differed significantly between BSA- and non-immunized fish, despite some sera from non-immunized fish with slight high OD values. These results suggest that the MAbs produced in this study reacted specifically with the IgM from olive flounder.

• Journal of fish pathology
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• v.37 no.1
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• pp.71-78
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• 2024

Streptococcosis, caused by Streptococcus iniae and Streptococcus parauberis is an important bacterial disease that affects in olive flounder in Korea. In Korea, multivalent bacterial vaccines are used to prevent streptococcal diseases in aquaculture. In this study, commercial vaccines containing formalin-inactivated bacterial cells of S. iniae and S. parauberis were administered at six fish farms and one unvaccinated fish farm were designated for investigation (Wando; 4 sites and Jeju; 3 sites). Blood was collected from vaccinated and unvaccinated olive flounders, and titers of antibodies against S. iniae and S. parauberis in serum were analyzed using ELISA. After a one shot vaccination in the farms at Jeju (farm A) and Wando (farm D), the proportion of individuals with specific antibodies against S. parauberis OD values of 0.4 or higher was 60% and 53.5%, respectively. But after booster vaccination, the proportion of individuals with serum OD values of 0.4 or higher was higher substantially increased to 96.6% (farm A) and 100% (farm D). The levels of S. parauberis specific antibodies of olive flounder were increased after vaccination in three fish farms (farm D, E, and F), but not S. iniae specific antibodies.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.497-503
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• 2016

This study aimed to optimize the culture conditions of recombinant Escherichia coli expressing otsBA using crude glycerol for the enhanced production of trehalose. The effects of culture temperature and isopropyl ${\beta}$-D-1-thiogalactopyranoside (IPTG)-induction were investigated. Trehalose production and cell growth were highest when cells were cultured at $37^{\circ}C$ and induced with IPTG. The concentrations of IPTG, validamycin A, and NaCl were optimized using Box-Behnken design. Statistical analyses of the experimental data revealed that the concentrations of IPTG and NaCl had significant effects on trehalose production, but that of validamycin A did not. Contour plot analysis and model calculation showed that the highest amount of trehalose could be produced at 298 mM NaCl and 0.1 mM IPTG. Under these optimal conditions, the optical density at 600 nm and trehalose production were $5.4{\pm}0.2$ and $304{\pm}15mg/l$, respectively.