• Microbiology and Biotechnology Letters
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• v.47 no.2
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• pp.250-258
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• 2019

Peniphora sp. JS17, isolated from forest old tree, produced extracellular enzymes that decolorized human hair melanin. The JS17 strain had laccase and manganese peroxidase activity while it did not has lignin peroxidase activity. Batch culture indicated that the melanin decolorization activity of JS17 strain originated from laccase. The culture conditions to maximize the production of melanin bleaching enzymes from Peniophora sp. JS17 mycelia were investigated. Among the tested media for the laccase production, minimal medium (2% glucose, 0.2% malt extract, 0.1% $KH_2PO_4$, 0.4% $MgSO_4{\cdot}7H_2O$) showed the highest activity of laccase. Then, to optimize the culture condition for the laccase activity, the influence of various carbon and nitrogen sources was investigated in minimal medium. Among various carbon and nitrogen sources, 2% xylose and 0.4% tryptone showed the highest production of laccase, respectively. The enzyme was purified using $(NH_4)_2SO_4$ precipitation and Hitrap Q sepharose column, and the purified enzyme showed two isoenzymatic bands with molecular masses of about 70 kDa by SDS-PAGE. The melanin decolorization activity was 77% and 55% within 48 h in the presence of 1-hydroxybenzotriazole (HBT) and syringaldehyde, respectively, whereas only about 9% melanin decolorized in case of no mediator.

• Korean Journal of Food Science and Technology
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• v.47 no.4
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• pp.438-445
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• 2015

The production of GABA was optimized by co-cultivation of immobilized Lactobacillus plantarum K154 (ILK) with Ceriporia lacerata cultures. The mycelial culture of C. lacerata was performed in a defined medium containing 3% glucose, 3% soybean flour, and 0.15% $MgSO_4$ in a submerged condition for 7 days at $25^{\circ}C$, resulting in the production of 29.7 g/L mycelia, 3.1 g/L exopolysaccharides, 2% (w/w) ${\beta}$-glucan, 68.96 unit/mL protease, and 10.37 unit/mL ${\alpha}$-amylase. ILK in C. lacerata culture showed viable cell counts of $3.13{\time}10^9CFU/mL$ for immobilized cells and $1.48{\time}10^8CFU/mL$ for free cells after 1 day. GABA production in the free and immobilized cells was 9.96 mg/mL and 6.30 mg/mL, respectively, after 7 days. A recycling test of ILK in the co-fermentation was consequently performed five times at $30^{\circ}C$ for 15 days, resulting in the highest production of GABA. GABA could also be efficiently overproduced by co-cultivation with the produced polysaccharides, ${\beta}$-glucan, peptides, and probiotics.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.13 no.11
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• pp.5179-5186
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• 2012

Miglitol, a well-known therapeutic intervention agents for diabetes, exhibits competitive inhibitory activity against ${\alpha}$-glucosidase and it is usually produced through three sequential steps including chemical and bioconversion processes. Gluconobactor oxydans (G. oxydans) belonging to acetic acid bacteria biologically, converts 1-deoxy-1-(2-hydroxyethylamino)-D-glucitol (P1) into a key intermidiate, 6-(2-hydroxyetyl) amino-6-deoxy-${\alpha}$-L-sorbofuranose (P2) by incomplete oxidation. In this study, we identified and optimized fermentation conditions of CK-2165, that was selected in soil samples by comparing the bioconversion yield. CK-2165 strain was found to be closely related to G. oxydans based on the result of phylogenetic analysis using 16S rDNA sequence. Utilization of API 20 kits revealed that this strain could use glucose, mannose, inositol, sorbitol, rhamnose, sucrose, melibiose, amygdalin and arabinose as carbon sources. The culture conditions were optimized for industrial production and several important factors affecting bioconversion rate were also tested using mycelial cake. Cell harvested at the late-stationary phase showed the highest bioconversion yield and $MgSO_4$ was critically required for the catalytic activity.

• Applied Biological Chemistry
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• v.39 no.3
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• pp.172-176
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• 1996

Effect of culture conditions such as pH, temperature, agitation speed and oxygen transfer rate on xylitol production from xylose by Candide parapsilosis ATCC 21019 mutant was investigated in a jar fermentor. The initial concentration of xylosr was fixed at 50 g/l in this experiment. When pH was increased, cell growth and xylose consumption rate were increased, but maximum xylitol production was shown in the range of pH 4.5 and 5.5 with a yield of 0.68 g/g-xylose. The optimal temperature for xylitol production was determined to be $30^{\circ}C$. Considering the importance of dissolved oxygen tension, for xylitol production, the effect of oxygen transfer rate coefficient $(k_La)$ on fermentation parameters was carefully evaluated in the range of $20{\sim}85\;hr{-1}\;of\;k_La$ (corresponding to $100{\sim}300$rpm of agitation speed). The xylitol production was maximized at $30\;hr^{-1}\;of\;k_La$(150 rpm). A higher oxygen transfer rate supported better cell growth with lower xylitol yield. It was determined that maximum xylitol concentration, xylitol yield and productivity was 35.8 g/l, 71.6% and $0.58\;g/l{\sim}hr^{-1}$, respectively, at $30\;hr^{-1}\;of\;k_La$ In order to further increase xylitol productivity, ferementation using the concentrated biomass(20 g/l) was carried out at the conditions of pH 4.5, $30^{\circ}C$ and $30\;hr\;1$ of oxygen transfer rate. The final xylitol concentration of 40 g/l was obtained at 18 hours of culture time. From this result, it was calculated that xylitol yield was 80ft on the basis of xylose consumption and volumetric productivity was $2.22\;g/l{\sim}hr$ which was increased by $3{\sim}4$ fold compared with $0.5{\sim}0.7\;g/l-hr$ obtained in a normal fermentation condition.

• Journal of Life Science
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• v.28 no.5
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• pp.577-586
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• 2018

We recently reported the potential probiotic properties of Lactobacillus brevis SBB07 isolated from blueberries. The present study investigates the effect of culture conditions such as temperature, initial pH, culture time, and medium constituent for industrial application. The ingredients of the medium to improve cell growth were selected by Plackett-Burman design (PBD) and central composite design (CCD) within a desirable range. The PBD was applied with 19 factors: seven carbon sources, six nitrogen sources, and six microelements. Protease peptone, corn steep powder (CSP), and yeast extract were found to be significant factors for the growth of SBB07. The CCD was then applied with three variables found from the PBD at five levels, and the optimum values were decided for the three variables: protease peptone, CSP, and yeast extract. In the case of the growth of SBB07, the proposed optimal media contained 2.0% protease peptone, 2.5% CSP, and 2.0% yeast extract, and the maximum dried-cell weight was predicted to be 2.93963 g/l. By the model verification, it was confirmed that the predicted and actual results are similar. Finally, the study investigated the effects of incubation temperature and initial pH at the optimized medium. It was confirmed that the dried-cell weight increased from $2.2933{\pm}0.0601g/l$ to $3.85{\pm}0.0265g/l$ when compared to the basal medium at $37^{\circ}C$ and initial pH 8.0. Establishing the optimal culture condition for SBB07 provides good potential for applications in probiotics and can serve as the foundation for the industrialization of materials.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1379-1385
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• 2017

The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by $^1H$ NMR, $^{13}C$ NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (${\beta}-sitosterol$ and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.388-393
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• 2017

We report an Agrobacterium-mediated transformation procedure optimized for 'Kyoho' that is a major table grapevine cultivar in Korea, and its transgenic plants with antifreeze protein gene of carrot (DcAFP). The full length of DcAFP coding region in accordance with the previous report was isolated from young leaves of carrot and recombined into a plant transformation vector. Ethylene inhibitors such as silver nitrate and aminoethoxyvinylglycin (AVG) supplemented in a co-cultivation medium distinctly increased frequency of shoot regeneration when explants were sub-cultured in a selection medium: particularly ten-fold higher in treatment with 0.1 mg/L AVG than one without ethylene inhibitor. Among various antibiotics and their concentrations, the combination of 150 mg/L cefotaxime plus 150 mg/L $Clavamox^{TM}$ was selected for elimination of Agrobacterium cells in addition to minimization of adverse effect on shoot regeneration, while 50 mg/L kanamycin monosulfate effectively suppressed regeneration of non-transgenic shoots. Applying the elucidated culture condition, we finally obtained a total of 5 transgenic 'Kyoho' plantlets with DcAFP, of which integration with the grapevine genome and transcription was confirmed by nucleic acid analyses.

• Journal of Plant Biotechnology
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• v.5 no.2
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• pp.83-86
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• 2003

A reverse transcription polymerase chain reaction (RT-PCR) protocol was developed using two specific 22-mer primers located in coat protein gene of SPFMV. A 411 bp PCR-product was detected in virus infected plants as well as tissue culture raised sweet potato but not in healthy plants. For optimization of RT-PCR protocol, the optimum crude nucleic acid concentration, annealing temperature, primer concentration and numbers of PCR-cycle for maximum sensitivity and specificity were determined. The optimum condition for RT-PCR was as follows: RT-PCR reaction mixture was one-step mixture, containing 50 pmol of primer, 30 units of reverse transcriptase, 5 units of RNasin, and the crude nucleic acid extracts (200 ng). In RT-PCR, cDNA was synthesized at $42^{\circ}C$ for 45 min before a quick incubation on ice after pre-denaturation at $95^{\circ}C$ for 5 min. The PCR reaction was carried out for 40 cycles at $96^{\circ}C$ for 30 see, $63^{\circ}C$ for 30 sec, $72^{\circ}C$ for 1 min, and finally at $72^{\circ}C$ for 10 min. The viral origin of the amplified product was confirmed by sequencing, with the sequence obtained having $95-98\%$ homology with published sequence data for SPFMV. The benefits of this RT-PCR based detection of SPFMV would be simple, rapid and specific.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.327-333
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• 1988

For optimal production of recombinant human interleukin-2 (IL-2) in E. coli the effect of fermentation conditions on cell growth, IL-2 production, and stability of recombinant cells were investigated. Among the complex nutrients tested in this work, yeast extract, peptone and corn steep liquor were found to be effective for recombinant cell growth. The recombinant cells were maintained stably under repression condition (3$0^{\circ}C$), but the stability of recombinant cells were drastically reduced upon induction of IL-2 expression (42$^{\circ}C$) even under the selection pressure. Addition of antibiotics to the culture medium resulted in the cell growth inhibition without significant improvement in recombinant stability. When the expression of IL-2 gene was induced at different growth phases, highest IL-2 production was achieved by the induction of IL-2 at the middle-exponential growth phase. It was found that the production of IL-2 significantly inhibited the cell growth and the ex-pression of other genes in the plasmid.

• Journal of Marine Bioscience and Biotechnology
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• v.11 no.1
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• pp.14-22
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• 2019

Omega-3 fatty acids and carotenoids, which are known as representative high-value substances derived from microalgae, are being studied from various diatoms. Most of the diatoms contain fucoxanthin and omega-3 fatty acid. Fucoxanthin produced by diatom has been reported as bioactive compounds exhibiting strong antioxidant, anticancer and anti-inflammatory activities. However, the low growth rate and fucoxanthin content of diatoms are one of the big obstacles to the industrial application. In this study, indigenous marine diatom Achnanthidium sp. BS-001 was isolated for a candidate of fucoxanthin producer. Light intensity and temperature for the culture of Achnanthidium sp. BS-001 were optimized on PhotoBiobox. Optimization of silicate concentration for increasing BS-001 biomass productivity was confirmed in F/2 medium with various concentration of sodium silicate. As a result, condition of light intensity, temperature, and silicate concentration for optimal cultivation were $150{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, $18^{\circ}C$ and 0.106 mM, respectively. Maximum biomass productivity reaches to $154.3mg{\cdot}L^{-1}{\cdot}day^{-1}$, and then the content of omega-3 fatty acids and fucoxanthin were $19.4mg{\cdot}g^{-1}$, $9.05mg{\cdot}g^{-1}$, respectively. These results indicate that Achnanthidium sp. BS-001 has the potential to be used as a source of omega-3 fatty acids and fucoxanthin.