• 대한치과보존학회:학술대회논문집
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• 대한치과보존학회 2003년도 제120회 추계학술대회 제 5차 한ㆍ일 치과보존학회 공동학술대회
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• pp.621-621
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• 2003

I. Objectives In order to examine the immunoresponse of host cells to Enterococcus faecalis, this in vitro study monitored the production of Interleukin-2(IL-2), Interleukin-4(IL-4) and Transforming growth $factor-{\beta}1(TGF-{\beta}1)$ in human lymphocytes. II. Materials and methods Enterococcus faecalis(ATCC29212) strains were used in this study. Strains were grown in 1-liter cultures in 85% N2-10% H2-5% $CO_2$chamber for 3 days at $37^{\circ}C$. The medium used was 3.7% brain heart infusion broth. Bacterial cells harvested from 1-liter cultures were washed, suspended in 20ml of phosphate-buffered saline(PBS). Suspensions of bacterial cells were disrupted by sonication on ice for 5 min. Protein concentration was determined by the Bicinchoninic acid(BCA) protein assay.(omitted)

• Mycobiology
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• 제36권3호
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• pp.148-151
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• 2008

A novel biomass was prepared from Pichia anomala KCCM 11473, which grew well in ginseng-steaming effluent (GSE), and its physiological functionalities and enzyme activities were determined. When the strain was cultured in the GSE (pH 6.0) at 30$^{\circ}C$ for 48 h, 1.6 mg of biomass per ml-cultures was produced. The cell-free extract of the biomass showed high antihypertensive angiotensin I-converting enzyme inhibitory activity of 72.0% and anticholesteromia HMG-CoA reductase inhibitory activity of 46.5%. The cell-free extract also showed 13.0 U per ml and 8.5 U per ml of neutral protease activity and alkaline protease, respectively.

• 한국미생물·생명공학회지
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• 제51권1호
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• pp.59-68
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• 2023

Xylanase is an industrially relevant enzyme used for the production of xylobiose and xylose. Various methods are used to enhance the microbial yield of xylanase. In the present study, co-culturing of Bacillus subtilis and Bacillus pumilus were investigated using submerged fermentation for xylanase production, which was markedly increased when sal, sagwan, newspaper, wheat bran, and xylan were used as single carbon sources. Maximum xylanase production was reported after 5 days of incubation in optimized media at pH 7.0 and 37℃, resulting in 2.69 ± 0.25 µmol/min by coculture. The 1:1 ratio of sal and sagwan in optimized production media was shown to be suitable for xylanase synthesis in submerged fermentation (SMF). In comparison to mono-culture using B. pumilus and B. subtilis, co-culturing resulted in an overall 3.8-fold and 2.15-fold increase in xylanase production, respectively.

• Journal of Plant Biotechnology
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• 제4권4호
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• pp.143-150
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• 2002

A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

• 한국약용작물학회지
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• 제8권3호
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• pp.250-258
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• 2000

약용식물의 기내 (in vitro) 배양에 의한 유용물질의 생산을 위하여 대황 (Rheum undulatum L.)의 유묘 조직 절편에 Agrobacterium rhizogenes A4을 감염시켜 형질전환을 유도하였다. 형질전환된 부정 근인 모상근은 WPM 배지에서 RCM 배지의 3.4배의 성장률 증가를 나타내었으며, 지표 물질인 tannin의 생산량 또한 동일 배지에서 가장 높았다. 배지내 식물성장조절물질의 처리에 의한 모상근의 성장 및 tannin 생산촉진 효과는 식물성장조절물질을 처리하지 않은 대조구에 비해 2 mg/L IAA 처리구와 0.5 mg/L ABA 처리구에서 각기 1.7배의 성장 (0.72 g dry weight/flask)과 1.4배의 tannin (0.67 mg/g fresh weight)의 생산량 증가를 가져왔다. 배지내 탄소원인 sucrose 는 3%와 5%에서 최대성장 (0.54 g dry weight/flask)과 tannin 생산 (0.46 mg/g fresh weight)을 보였으며, chitosan의 농도는 50 mg/L 처리구에서 tannin의 생합성에 영향을 주었다.

• Food Science and Biotechnology
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• 제18권6호
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• pp.1423-1429
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• 2009

Rhizopus oryzae KSD-815 was isolated from nuruk that has been used to make Korean traditional wines. This study was performed to investigate the effect of cultures of R. oryzae KSD-815 on cardiovascular disorders and cancer metastasis. Firstly, these cultures were sequentially fractionationed with n-hexane (TAHe), ethylacetate (TAE), n-butanol (TAB), and $H_2O$ (TAW). The TAE inhibited the activity of angiotensin-converting enzyme (ACE) and TAB suppressed platelet aggregation in vitro. TAE and TAB inhibited cell motility of human breast cancer cells. Furthermore, TAW interrupted the formation of neovasculature and tube-like structure, and down-regulated the expression of angiogenic factors, basic fibroblast growth factor (bFGF), tumor necrosis factor-$\alpha$ (TNF-$\alpha$), and hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) in breast cancer cells. These results indicated that cultures of R. oryzae KSD-815 display the inhibitory activities on hypertension, platelet aggregation, and metastasis, and suggest that these cultures might be further probed for the purposes as therapeutic agents or dietary supplements.

• 한국가축번식학회지
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• 제22권1호
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• pp.43-50
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• 1998

The effect of oxygen tension on embryonic development in co-culture was evaluated from the standpoint of the reduction of dissolved oxygen concentration by the oxygen consumption of feeder cells. Three co-culture systems using bovine oviductal epitherial cells (BOEC), African green monkey kidney cells (Vero cells) or buffalo rat liver cells (BRLC) have been compared in terms of development of bovine embryos derived from oocytes matured and fertilized in vitro. Among the co-cultured embryo, Vero cells su, pp.rted the highest developmental rate (29%) and the other two showed the similar rates. When the co-cultures were incubated in three different oxygen tension such as 5, 10, 20% oxygen atmosphere, embryos co-cultured with Vero cells at 10%-O2 resulted in the highest percentage of development. From the measurement of oxygen consumption of feeder cells, BRLC consumed 1.38　10-10 mg-O2/min/cell which was higher than 0.94　10-10 and 0.26　10-10mg-O2/min/cell for Vero cells and BOEC, respectively. Based on the oxygen consumption data, the phenomena of optimum oxygen tension required in embryo development in vitro has been analyzed, and we suggested that gas phase oxygen concentration, oxygen consumption rate of feeder cells and the number of feeder cells should be considered for the design of optimal co-culture system for effective fertilization of embryos in vitro.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.451-458
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• 2013

The human gastric pathogen Helicobacter pylori (Hp) has been considered a microaerophile. However, we recently reported that, when supplied with 10% $CO_2$, Hp growth is stimulated by an atmospheric level of $O_2$, suggesting that Hp is a capnophilic aerobe. In this study, we investigated the effects of aerobic $O_2$ tension on Hp cells by comparing gene expression profiles of cultures grown under microaerobic and aerobic conditions in the presence of 10% $CO_2$. The results showed that overall differences in gene expression in Hp cells grown under the two $O_2$ conditions were predominantly growth-phase-dependent. At 6 h, numerous genes were down-regulated under the aerobic condition, accounting for our previous observation that Hp growth was retarded under this condition. At 36 h, however, diverse groups of genes involved in energy metabolism, cellular processes, transport, and cell envelope synthesis were highly up- or down-regulated under the aerobic condition, indicating a progression of the cultures from the log phase to the stationary phase. The expression of several oxidative stress-associated genes including tagD, katA, and rocF was induced in response to aerobic $O_2$ level, whereas trxA, trxB, and ahpC remained unchanged. Altogether, these data demonstrate that aerobic $O_2$ tension is not detrimental to Hp cells but stimulates Hp growth, supporting our previous finding that Hp may be an aerobic bacterium that requires a high $CO_2$ level for its growth.

• Journal of the Korean Wood Science and Technology
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• 제47권2호
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• pp.210-220
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• 2019

This work aimed to evaluate the influence of culture conditions on laccase production in the co-culture of wood-rotting fungus with Trichoderma sp. The effects of infection extent, infection time, and culture filtrate of Trichoderma sp. on the laccase production by wood-rotting fungus in co-culture were examined. T. rubrum LKY-7 and T. longibrachiatum were selected as fungi which are effective in co-culture for laccase production. A significant increase in laccase activity was observed when T. rubrum LKY-7 was co-cultured with T. longibrachiatum in glucose-peptone liquid medium, yielding an increase of more than 5 times in laccase activity, as compared with control. Laccase production by T. rubrum LKY-7 during co-culturing was significantly influenced by the infection extent and the infection time of T. longibrachiatum. Maximal laccase activity was obtained when T. rubrum LKY-7 culture was infected by T. longibrachiatum after 3 days of cultivation at an inoculum size ratio of 0.5 to 1. The addition of culture filtrate or autoclaved mycelium of T. longibrachiatum to T. rubrum LKY-7 culture did not significantly enhance laccase production by T. rubrum LKY-7 as compared with control (mono cultures of T. rubrum LKY-7).

• Asian-Australasian Journal of Animal Sciences
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• 제18권10호
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• pp.1425-1429
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• 2005

Megaspahera elsdenii YJ-4, which was previously isolated as a producer of trans-10, cis-12 CLA, was studied for its carbon source on the CLA production. M. elsdenii YJ-4, was incubated with glucose and lactose, and cultured in batch and continuous culture systems with linoleic acid at various pHs to investigate CLA production. Batch cultures of the ruminal bacterium, M. elsdenii YJ-4, were resistant to stearic acid and linoleic acid, and little growth inhibition was observed even when the fatty acid concentration in the culture was as much as 4 mg $ml^{-1}$. Stationary phase batch cultures (0.25 mg bacterial protein $ml^{-1}$) that had been grown on lactate and incubated with linoleic acid (0.20 mg $ml^{-1}$) produced approximately 12 ${\mu}g$ trans-10, cis-12 CLA mg $protein^{-1}$ and little cis-9, trans-11 CLA was detected. Some linoleic acid was converted to hydrogenated products (chiefly stearic acid), but these fatty acids were less than 5 ${\mu}g$ mg bacterial $protein^{-1}$. Stationary phase batch cultures that had been grown on glucose produced at least 3-fold less trans-10, cis-12 CLA than ones grown on lactate. Cells from lactate-limited continuous cultures produced less trans-10, cis-12 CLA than those from batch culture, but only if the pH was greater than 6.4. When the pH of the lactate-limited continuous cultures was lower than 6.4, trans-10, cis-12 CLA and hydrogenated products declined. Cells from glucose-limited continuous cultures produced less trans-10, cis-12 CLA and hydrogenated products than the cells that had been limited by lactate, but pH had little impact on this production. These results support the idea that M. elsdenii YJ-4 could be one of the major producers of trans-10, cis-12 CLA which causes cows to produce milk with a low fat content.