• 생명과학회지
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• 제12권5호
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• pp.566-569
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• 2002

Acetobacter strains는 포도당을 함유한 복합배지에서 셀룰로오스를 합성할 수 있는 세균이다. 본 연구에서는 Acetobacter sp. A9에 의한 셀룰로오스 생산에 영향을 미치는 몇 가지 배양 환경요인을 진탕배양을 통하여 조사하였다. 배지에 3-(N-morpholino) propanesulfonir acid (MOPS)와 CaCO$_3$ 등과 같은 완충성분의 첨가는 hcetobacter sp. A9에 의한 셀룰로오스 생산량을 증가시켰다. 따라서 배양시간 경과에 따른 배양액의 수소이온농도 조절이 경제적인 셀룰로오스 대량생산에 중요한 요인임을 알 수 있었다. 배지에 직경 10 mm의 bead를 첨가함으로써 다양한 기능을 가진 'disintegrated bacterial cellulose'가 생산되었다.

• ALGAE
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• 제32권3호
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• pp.245-259
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• 2017

We report on the culture growth and stress-induced secondary carotenogenesis in a biotechnologically promising but largely unexplored chlorophyte Coelastrella rubescens strain Vinatzer/Innsbruck V 195. Changes in the cell morphometry, biomass accumulation, its carotenoid and fatty acid profiles were followed in the cultures supplemented with either inorganic ($CO_2$) or organic (sodium acetate) carbon on the background of low-pH stress. Collectively, the results of the study characterize C. rubescens as a biotechnologically promising, potentially double-purpose organism. It produces several secondary keto-carotenoids with a considerable proportion of astaxanthin and canthaxanthin. At the same time, the cell lipid fatty acid profile of this microalga is suitable for obtaining a high-quality biodiesel complying with the strictest EN14214 European standard.

• Journal of Applied Biological Chemistry
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• 제43권3호
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• pp.136-140
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• 2000

A novel 6-deoxyhexose biosynthetic gene cluster different from the one for the biosynthesis of streptomycin was isolated from Streptomyces griseus using specifically designed PCR primers to compare the sequence of known dTDP-glucose synthase genes. We cloned a 5.8-kb DNA from Streptomyces griseus ATCC10137, which contained the 4-ketoreductase homologue (grsB), dTDP-glucose synthase (grsD), and dTDP-glucose 4, 6-dehydratase (grsE) genes. Escherichia coli cultures containing plasmid of the PCR product which encoded the grsE region under the controUed T7 promoter were able to catalyze the formation of dTDP-4-keto-6-deoxy-D-glucose from TDP-glucose. The enzyme showed high substrate specificity, being specific to only dTDP-glucose that is known to be incorporated into secondary metabolites such as antibiotics.

• Journal of Microbiology and Biotechnology
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• 제23권9호
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• pp.1327-1338
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• 2013

The humanized anti-hepatocyte growth factor (HGF) monoclonal antibody (mAb) YYB-101 is a promising therapeutic candidate for treating various cancers. In this study, we developed a bioprocess for large-scale production of YYB-101 and evaluated its therapeutic potential for tumor treatment using a xenograft mouse model. By screening diverse chemically defined basal media formulations and by assessing the effects of various feed supplements and feeding schedules on cell growth and antibody production, we established an optimal medium and feeding method to produce 757 mg/l of YYB-101 in flask cultures, representing a 7.5-fold increase in titer compared with that obtained under non-optimized conditions. The optimal dissolved oxygen concentration for antibody production was 70% $pO_2$. A pH shift from 7.2 to 7.0, rather than controlled pH of either 7.0 or 7.2, resulted in productivity improvement in 5 L and 200 L bioreactors, yielding 737 and 830 mg/ml of YYB-101, respectively. The YYB-101 mAb highly purified by affinity chromatography using a Protein A column and two-step ion exchange chromatography effectively neutralized HGF in a cell-based assay and showed potent tumor suppression activity in a mouse xenograft model established with human glioblastoma cells.

• KSBB Journal
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• 제31권4호
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• pp.270-276
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• 2016

In glycoprotein, Terminal sialic acid residues of N-linked glycan are imperative things because they prevent the recognition from asialoglycoprotein-receptor that affect the half-life of glycoproteins. So establishment of culture process for enhancing sialic acid is important to maximize sialic acid contents of glycoprotein. In this study, we investigated effects of biphasic culture of Chinese hamster ovary (CHO) cells producing albumin-erythropoietin to increase sialylation. Biphasic cultures were performed with shift of $CO_2$ concentrations and temperatures at day 5 when viable cell density was decreased and sialidase was started to be released by cell lysis. The examined temperature set points were 33, 35 and $37^{\circ}C$ respectively and the $CO_2$ concentration was 1, 5, 10 and 15%. We confirmed that sialidase activity was the lowest in biphasic culture that was shifted from normal culture condition to 1% of $CO_2$ and $33^{\circ}C$ on day 5. However, the temperature and concentration of $CO_2$ have little effect on activity of ${\alpha}2,3$-sialyltransferase. Also, sialic acid contents were enhanced 1.13-fold higher than that in control culture. In conclusion, Biphasic cultivation in CHO cells led to inhibition of sialidase activity and increases of sialylated glycan.

• 대한환경공학회지
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• 제33권5호
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• pp.314-324
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• 2011

하수를 이용해서 배양된 조류는 바이오디젤 생산에 유용한 자원이다. 그러나 실제 하수에서 조류의 영양염류 신진대사와 하수 세균과의 상호작용에 관한 연구는 미흡하다. 본 연구에서는 하수로 배양되는 대표적 조류균주인 Ankistrodesmus gracilis SAG 278-2에 의한 하수 내 질소, 인 제거 거동을 평가하였고, 조류와 상호작용하는 하수 내의 세균 군집을 분석하였다. 하수 슬러지 세균 군집과 비교하였을 때, 조류-세균 복합 군집은 하수 내보다 높은 영양염류 제거를 보였다. 16S rRNA 유전자 분석 결과, 조류-세균 군집에서 조류가 성장함에 따라 Unclassified Alcaligenaceae 세균이 선택적으로 우점됨을 알 수 있었고, 조류에 의해서 선택적으로 우점화된 하수세균은 자연 수질 환경에서 조류와 공생적으로 상호작용 하는 것으로 알려진 Alcaligenes faecalis subsp. 5659-H와 계통학적으로 가까운 것으로 밝혀졌다. 본 연구의 결과, 하수 내의 높은 영양염류 제거를 보이는 조류-세균 복합 군집에서의 조류의 성장 및 신진대사가 특정 세균의 분포에 영향을 주는 것을 알 수 있었다.

• Clinical and Experimental Reproductive Medicine
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• 제19권2호
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• pp.133-141
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• 1992

We examined effects of co-culture with human oviduct epithelial cells (HOEC) on the development of mouse and human embryos from early embryonic· stage to late morula or blastocyst stage (LM or B). In human, embryos were transferred and pregnancy rate was investigated. The HOEC, collected from surgically removed fallopian tube, were cultured in medium-199 supplemented with 20 % fetal cord serum (FCS). The HOEC were characterized by using immunocytochemical staining with anticytokeratin antibody and then used for cultures of mouse and human embryos. Results obtained from co-culture system were as follows. Development rate of mouse embryos was improved by co-culture system at late developmental stage (p<0.025). Human supernumerary embryos remained after transfer, unsuitable for freezing because of their poor quality, were co-cultured for 72hrs. Co-culture (78.79%) or conditioned medium (78.26%) system improved the developmemt rate, significantly, in comparision with control (11.11%)(p<0.00l). Co-cultured (85.71%) human zygotes for 24hrs showed the better development rate in comparision with control (50.00%) (p<0.01). When we transferred embryos cultured with the HOEC to patients, we obtained one pregnancy. Co-cultured human zygotes for 24hrs showed the better quality and viability for the replacement in comparision with control (p<0.01). In addition, improved pregnancy rate was obtained. Our results suggest that the co-culture system can rescue early degenerating embryos by improving early development and yield a resonable number of blastocyst for the appropriate replacement. The effect provided by cultured HOEC is not species specific for the development of embryos and it can be used to overcome in vitro blocks for the development. And also the co-culture system offers the possibility to freeze embryos at blastocyst stage which is more sucessful stage for the freezing. The HOEC monolayer may provide some stimulus via specific factor, which is unknown, to the development of embryos. Our results showed that the co-culture system with HOEC can be an alternative to conventional culture system.

• 한국동물위생학회지
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• 제23권1호
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• pp.45-59
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• 2000

The present study was conducted to investigate the epidemiological properties of salmonellosis of poultry farms in Kyongbuk province during the relied from November 1998 to November 1999 And antibiotic susceptibility and biochemical characteristics of 120 Salmonella cultures isolated from chicken samples were also investigated. The results obtained through this study were summarized as follows, 1. Among 667,200 chickens of 31 flocks in 17 layer farms and 9 broiler farms, 61,350 chickens of 31 flocks were died with salmonellosis. The death rate of 25 farms varied from 0.1% to 75.0%, and the average death rate was 9.2%. 2. According to etiological agents, fowl typhoid was shown the most predominant outbreak among the salmonellosis during a year, which accounted for 8R.0%(22/25) of the total case 3. The serotypes of 120 Salmonella isolates were identified as 7 strains(5.8%) of S pullorum, 10 strains(8.4%) of S typhimurium and 103 strains(85.8%) of S gallinarum. 4. Most outbreak of fowl typhoid were prevalent on the layer chicken farms(77.1%), and the summer season(45.5%) also appeared the most hazardous season during the year. 5. It seemed that the Hyline breed(70.6%) was the most susceptible among the layer chicken breeds, and followed by Isabrown(23.5%), Tetra(5.9%) in order. 6. In layers, 76.4% of fowl typhoid occurred commonly from 14 to 40 weeks including the early laying peroid, but in broiler farms, all cases was outbreak within first second weeks. 7 All the strains of S pullorum were resistant to lincomycin(Lm), penicillin(Pm), and steptomycin(Sm), but sensitive to amikacin(Ak), ampicillin(Am), cephalothin(Ce), ciprofloxacin (CiP), chloramphenicol(Cm), colistin(Co), enrofloxacin(Enr), furazolidone(Fu), gentamicin(Gm), kanamycin(Km), neomycin(Nm), polymyxin(Po), and teracycline(Tc). All the strains of S typhimurium were resistant to Lm(100%), Pm(100%), Po(90%), and Sm(90%), but were sensitive to Ak, Am, Ce, CiP, Cm, Co, Enr, Fu, Gm, Km, Nm, and Tc. 8. Minimum inhibitory concentration(MIC) of 103 strains of S gallinarum were also evaluated and their patterns were much more variable than others. All the strains of S gallinarum were sensitive to Ak, Am, Ce, Cip, Cm, Enr, Fu, Km, and Nm, but resistant to Lm(100%), and Sm(100%), 99(96.1%) to Co, 83(80.6%) to Pm, and 83(80.6%) to Po, 55(53.4%) to Gm, and 33(32.0%) to Tc. 9. The multiple drug resistance patterns of 120 Salmonella strains were CoLmPmPo Sm pattern(34.2%), CoGmLmPmPoSmTc(20.8%), CoGmLmPmPoSm(13.3%), CoGmLmSm(7.5%), LmPmSm(7.5%), LmPm(6.7%), CoLmSmTc(3.3%),, CoGmLmSmTc(1.7%), GmLmSmTc(1.7%), CoGmLmPoSm(0.1%), LmPmPo(0.1%), CoLm Sm(0.1%), and LmSm(0.1%), in order.

• 대한임상검사과학회지
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• 제52권2호
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• pp.150-157
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• 2020

본 연구는 이산화망간(MnO₂)의 신경독성과 이에 대한 노박 덩굴 추출물의 보호 효과를 C6 glioma 세포를 배양하여 조사하였다. 본 연구를 위하여 세포생존율을 비롯하여 지질산화(LP) 억제능 및 XO 저해능과 같은 항산화 분석을 시행하였다. 그 결과 MnO₂는 배양 세포에 처리한 농도에 비례하여 세포생존율을 유의하게 감소시켰으며, 이 과정에서 XTT₅₀값은 146.7 μM로 나타나 Borenfreund and Puerner의 독성판정기준에 따라 중간독성(mid-cytotoxic)인 것으로 나타났다. 또한, 항산화제의 일종인 kaempferol (KAE)은 MnO₂의 독성에 의해 손상받은 세포의 생존율을 유의하게 증가시킴으로써 MnO₂의 독성을 방어하였다. 한편, MnO₂ 독성에 대한 노박덩굴(CO) 추출물의 영향 조사에 있어서, CO 추출물은 배양 세포에 MnO₂만을 처리한 것에 비하여 세포생존율을 유의하게 증가시킴으로써 MnO₂의 독성을 효과적으로 방어하였다. 이와 동시에 높은 LP 억제능과 XO 저해능과 같은 항산화 효과를 보여주었다. 이상의 결과에서 MnO₂의 독성에 산화적 손상이 관여하고 있음을 알 수 있었으며, CO 추출물은 LP 억제능과 XO 저해능과 같은 항산화능에 의하여 MnO₂의 독성을 효과적으로 방어한 것을 확인였다. 따라서, CO 추출물과 같은 천연소재는 파킨슨병 유발제인 MnO₂와 같이 산화적 손상과 관련된 질환적 중금속 독성을 경감 내지는 개선할 수 있는 유용한 물질이라 생각된다.

• IMMUNE NETWORK
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• 제11권5호
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• pp.281-287
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• 2011

Background: Biodegradable polymers have increasingly been recognized for various biological applications in recent years. Here we examined the immunostimulatory activities of the novel poly(aspartic acid) conjugated with L-lysine (PLA). Methods: PLA was synthesized by conjugating L-lysine to aspartic acid polymer. PLA-nanoliposomes (PLA-NLs) were prepared from PLA using a microfluidizer. The immunostimulatory activities of PLA-NLs were examined in mouse bone marrow-derived dendritic cells (BM-DCs). Results: PLA-NLs increased the number of BM-DCs when added to cultures of GM-CSF-induced DC generation on day 4 after the initiation of cultures. Examination of the phenotypic properties showed that BM-DCs generated in the presence of PLA-NLs are more mature in terms of the expression of MHC class II molecules and major co-stimulatory molecules than BM-DCs generated in the absence of PLA-NLs. In addition, the BM-DCs exhibited enhanced capability to produce cytokines, such as IL-6, IL-12, TNF-${\alpha}$ and IL-$1{\beta}$. Allogeneic mixed lymphocyte reactions also confirmed that the BMDCs were more stimulatory on allogeneic T cells. PLA- NL also induced further growth of immature BM-DCs that were harvested on day 8. Conclusion: These results show that PLA-NLs induce the generation and functional activities of BM-DCs, and suggest that PLA-NLs could be immunostimulating agents that target DCs.