Background: The objective of this study were to determine the antibacterial activity and antibiotic activity-enhancing effect of 70% ethanol extract of the root of Rumex japonicus Houtt. and its fractions when used in combination with gentamicin against aerobic skin flora. Methods and Results: The antibacterial activity and antibiotic (gentamicin) activity enhancing effect against aerobic skin flora were determined using the disc diffusion assay. Chloroform fraction (CF) and ethyl acetate fraction (EF) showed higher activities against Staphylococcus aureus and Staphylococcus epidermidis than those shown by other fractions. Regarding the antibiotic (gentamicin) activity-enhancing effect against aerobic skin flora, the n-hexane fraction (HF) and CF showed strong activity. The combination of HF and CF with gentamicin was evaluated using the broth dilution assay to determine the inhibitory effect on the growth of aerobic skin flora. The combination of CF with gentamicin exhibited the highest inhibitory effect on the growth of S. aureus and S. epdermidis. MTT assay performed to determine the viability of L929 cells revealed that EF treatment resulted in viability of 33.96 - 116.76% at the tested concentration. The combination of 70% ethanol extract and its other fractions with gentamicin showed low cell toxicity. Conclusions: Appropriate use of antimicrobial agents is important prior to the development of new antibiotics. The 70% ethanol extract of the root of R. japonicus Houtt. and its fractions showed significant synergism with gentamicin when used in combination against S. aureus and S. epdermidis. Thus, R. japonicus Houtt. could be used as a functional materials in antimicrobial-related fields.

Background: We investigated the antioxidative and tyrosinase inhibitory activities of 70% ethanol extract, and its fractions, of the root of Rumex japonicus HOUTT. Methods and Results: The total phenolic compound contents of the 70% ethanol extract and ethyl acetate fraction were 168.99㎎/ g and 651.78㎎/g, respectively. The antioxidant activity was compared through the DPPH radical and nitric oxide (NO) scavenging assays. The ethyl acetate fraction showed the highest DPPH radical and NO scavenging abilities, which confirmed the antioxidant activity. Specifically, the ethyl acetate fraction showed a higher DPPH radical scavenging ability than ascorbic acid. These results were related to the total phenolic compound content of the ethyl acetate fraction. Moreover, in the tyrosinase inhibition assay, the ethyl acetate fraction exhibited stronger inhibitory activity than arbutin, which was used as the positive control. The cell viability of L929 cells was analyzed by MTT assay after treatment with 70% ethanol extract and all fractions; no changes in viability were observed, which demonstrated the nontoxic nature of the extract and fractions. Conclusions: These results suggested that the extract from the root of R. japonicus and its ethyl acetate fraction could be a novel resource for the development of a cosmetic with antioxidant and tyrosinase inhibitory activity.

Background: This study aimed to investigate the antioxidat and antimicrobial activities of the methanol extract and its fractions prepared from the roots of Sanguisorba officinalis L. Methods and Results: The antioxidant activities were compared by evaluating the DPPH radical and nitric oxide (NO) scavenging ability. Measurement of DPPH radical scavenging ability showed that the SC50 values of the ethyl acetate fraction was 3.85 ㎍/㎖. The ethyl acetate fraction exhibited the most effective DPPH radical scavenging ability compared with the other samples. As for the NO scavenging ability, at all tested concentrations, the ethyl acetate fraction showed a higher scavenging activity than that of the extract and other fractions. These results are related to the total phenolic compound and flavonoid contents of the ethyl acetate fraction. Antimicrobial activity against foodborne pathogens was investigated using the disc diffusion assay. The ethyl acetate fraction showed the highest antimicrobial activity against gram-positive Staphylococcus aureus and Bacillus cereus. However, the chloroform fraction had a higher antimicrobial activity against gram-negative Vibrio vulnificus than that of the extract and other fractions. Conclusions: The results show that the ethyl acetate fraction had a higher antioxidant as well as antimicrobial activity, than did the other samples. Therefore, the ethyl acetate fraction has potential application in the food industry.

Background: In this study, we investigated the antibacterial and nitric oxide (NO) production inhibitory activities of 75% ethanol extract of Prunus sargentii branches and its fractions against acne pathogens. Methods and Results: The antibacterial activity against acne causing pathogens was determined using the disc diffusion assay. The ethyl acetate fraction showed higher activities against Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis than those shown by other fractions. In the DPPH radical and NO scavenging assays, the butanol fraction showed strong DPPH radical and NO scavenging abilities. These activities were related to the total polyphenol and flavonoid contents of butanol fraction. On the other hand, the chloroform and ethyl acetate fractions exhibited the highest NO production inhibitory activity in Lipopolysaccharide (LPS)-stimulated Raw 264.7 cells compared to those exhibited by other fractions. Conclusions: The extract and its ethyl acetate fraction from the branches of P. sargentii exhibited antibacterial activity and could be used as functional materials in antimicrobial related fields. Moreover, the chloroform and ethyl acetate fractions are potential antiinflammatory agents and butanol fraction acts as an effective radical scavenger.

This study was carried out to investigate the antibacterial activity against pathogens of acne and the anti-inflammatory effect of 75% ethanol extract and its fractions from the leaves of Prunus sargentii. In the antibacterial activity by the disc diffusion assay, the extract showed the highest effect against Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis in 5 mg/disc. However, the ethyl acetate fraction showed the highest antibacterial activity in 1 mg/disc. On the other hand, the hexane and chloroform fraction showed strong nitric oxide (NO) production inhibitory effect in lipopolysaccharide (LPS)-stimulated Raw 264.7 cell. In the cell viability of Raw 264.7 by MTT assay, the extract and all fractions were exhibited normal viabilities as nontoxic result. Consequently, the extract from the leaves of P. sargentii and its ethyl acetate fraction could be applicable to functional materials for antibacterial activity related fields. Moreover, the hexane and chloroform fraction could be applicable to candidate materials as anti-inflammatory agent.

In this study, we investigated the antibacterial activity, antioxidative activity and whitening effect of 75% ethanol extracts from different parts of Prunus sargentii. The total phenolic compound content of the branch extract was 277.92 mg/g as the highest level. In the measurement of DPPH radical scavenging ability, SC50 values of the cork layer and branch extract were 26.79 μg/ml and 30.13 μg/ml. In nitric oxide (NO) scavenging ability, SC50 values of the branch and leaf extract were 49.19 μg/ml and 55.55 μg/ml. All extracts exhibited higher NO scavenging ability than ascorbic acid used as positive control. On the other hand, in antibacterial activity against Staphylococcus epidermidis and Staphylococcus aureus by disc diffusion assay, the pure bark extract showed the highest activity. Moreover, tyrosinase inhibitory activity of cork layer, pure bark and branch extracts showed higher activity than arbutin used as positive control. In the cytotoxicity measurement by MTT assay, leaf extract was exhibited Raw 264.7 cell viabilities of 44.68~61.83% as cytotoxic result in tested concentration. In conclusion, the branch extract of Prunus sargentii will be a functional materials without damage compared to other parts such as pure bark or cork layer in the plant.

This experiment was carried out to obtain the cytotoxicity and antioxidative activity of Artemisiae Argi Folium. The total polyphenol contents in the ethyl acetate fraction of the ethanol extract and the methanol extract were 430.27mg/g and 427.84mg/g, respectively. In DPPH radical scavenging ability, SC50 values of the ethyl acetate fraction of the ethanol extract and the methanol extract were 32.64 μg/ml and 27.70 μg/ml as the same level of statistical with ascorbic acid. In the cytotoxicity measurement by MTT assay, the chloroform and hexane fraction, and each extract were exhibited higher cytotoxicity than the other fractions. In particular, the ethyl acetate fractions appeared high activity in DPPH radical scavenging ability were began to show cytotoxicity in 125 μg/ml. As a result, the ethyl acetate fraction of Artemisiae Argi Folium extract was the most highly active fraction in antioxidative activity. However, for the use of extracts and fractions from Artemisiae Argi Folium to related fields, the setting of appropriate concentration is required.

미생물 및 동물에 비해 식물에서는 혈전용해효소에 대한 연구가 매우 부족한 실정이며, 기존의 혈전용해효소가 가지는 혈전에 대한 비특이적, 부작용, 고가 등의 단점을 해결할 수 있는 새로운 혈전용해효소의 개발을 위하여 동백의 종자, 종피, 유엽 그리고 성엽으로부터 추출된 수용성 단백질의 혈전용해활성을 조사하였다. 각각의 동백부위로부터 추출된 조효소 용액은 기존 혈전 용해효소인 plasmin과 양성 대조군으로 하여 비교하여 fibrin agarose plate로 확인한 결과 fibrin clot을 효과적으로 분해하였다. 그 중 단백질 분해효소의 활성이 다른 부위보다 20-33배로 높았던 동백종자와 종피의 수용성 추출물의 혈전용해활성은 양성 대조군인 plasmin과 비교하여 1.6-2.0배의 높은 활성을 나타내었다. 전체 수용성 단백질은 30-80%황산암모늄을 이용하여 농축하였으며 혈전용해효소는 fibrin zymography를 수행하여 확인하였다. SDS-PAGE에 의하여 동백유엽의 혈전용해효소 분자량을 측정한 결과 45 kDa으로 단일 polypeptide임을 확인하였으며, 각종 pretense 저해제에 의한 영향을 조사한 결과 PMSF,그리고 TLCK에 강력하게 저해되는 것으로 보아 동백유엽의 혈전용해효소는 trypsin과 유사한 serine protease의 하나로 생각되었다. 그러나 EDTA와 DTT처리에 의해서는 효소활성의 저해가 두드러지게 나타나지 않고 오히려 증진된 양상을 확인할 수 있었다. 또한, 효소 활성에 미치는 pH 및 온도의 효과는 약간의 산성쪽에 가까운 pH 5.5와 30℃에서는 최적의 활성을 나타내었으며, 45℃ 이상의 온도에서는 효소활성이 급격히 감소하였다. 이상의 모든 결과로 볼 때 동백유엽의 혈전용해효소는 trypsin과 유사한 serine protease에 속하는 혈전용해효소임을 확인할 수 있었다.

기내배양을 통해 동백종자의 자엽절편으로부터 직접 체세포분화를 통하여 식물체를 재분화 시켰다. 종피를 제거한 후 표면살균 처리한 자엽절편을 2.4-D, NAA 및 BA가 단독 또는 조합 첨가된 MS 고체배지에 치상하였다. 원배체 형성을 통한 부정아 형성은 0.1 mg/l 2.4-D와 1 mg/l BA가 첨가된 처리구에서 이루어졌다. 부정아는 주로 자엽절편체의 배축에 가까운 기저부위에서 캘러스의 형성없이 직접 형성되었으며, 이로부터 형성된 유묘는 식물생장조절물질이 첨가되지 않은 기본배지에서 정상적인 개체로 발달 하였다. 이와같은 연구결과는 앞으로 동백나무의 육종과 산업적 이용에 필요한 형질전환 연구에 효과적으로 활용될 수 있을 것으로 사료된다.

공기주입형 생물반응기를 이용하여 오미자 현탁배양세포로부터 Gomisin J의 생산 가능성을 확인 하였다. 시료의 초기접종 농도와 탄소원의 농도에 따른 세포증식과 Gomisin J의 생산성은 6% sucrose가 첨가된 MB5배지 에 100 g (FCW)의 현탁배양세포를 접종하였을 경우 세포중량 (dry biomass)과 Gomisin J의 함량은 각각 43.5 g DCW/과 0.71 × 10-3 μg/g DCW을 얻을 수 있었다. 배양기내 산소의 공급량은 0.5로 공급해 주는게 바람직 할 것으로 사료되었다.

Transformed roots of Schisandra chinensis were obtained following co-cultivation of in vitro cultivated plantlet segments with Agrobaterium rhizogens ATCC15834. This root was examined for its growth and gomisin J contents under various culture conditions. Among the six basal culture media tested, WPM (Lloyd & McCown, 1980) medium supplemented with 5% sucrose was the best roots growth 6.2 (g D.W/flask) and gomisin J accumulation 1.56 (X10-3 ug/g D.W). Initial inoculum size correlated with the yield of biomass while gomisin J contents was not affect. Gomisin J production was influenced by the initial sucrose concentration and the highest production yield was achieved at the concentration of 7%. The optimal shaking speeds for roots growth and gomisin J production was 120 and 140 rpm, respectively.

Cell growth and gomisin J production by suspension cultures of Schisandra chinensis Baillon were investigated under various culture media, initial sucrose concentrations, shaking speeds, and inoculum sizes. Callus was induced from in vitro cultivated leaf segments on MS medium supplemented with 1 mg/l NAA. The maximum dry cell weight of 2.23 g was obtained at inoculum size of 0.5 g fresh cell weight and in MB5 medium supplemented with 1 mg/l NAA, 3% sucrose after 8 weeks. The production of gomisin J in suspension cell cultures was maximized in WPM medium containing 5% sucrose. The shaking speed for maintaining maximal cell dry weight was 100 rpm while the best shaking speed for gomisin J accumulation was 140 rpm.

동백나무의 잎과 꽃의 식용자원화를 위한 일환으로서 동백의 부위별 성분분석, DPPH radical 소거능에 의한 항산화활성, 및 항미생물 활성을 조사하였다. 수분 함량은 꽃과 어린 잎 부위에서 70% 이상의 함유율을 보였고, 조단백질의 함량은 꽃을 제외한 나머지 부위에서 3%이상이었으며, 지방 함량은 씨부위에서 23.08%로 많이 함유되어 있었다. 회분의 함량은 수피 부위에서 2.97%와 탄수화물의 함량은 에서 높게 나타났다. 항산화활성은 어린잎(RC50=30.37μg/mL), 꽃봉오리 (RC50=52.97μg/mL)와 꽃(RC50=59.48 μg/mL)이 높은 활성을 보였으며 합성 항산화제인 BHT의 RC50 값이 584.04μg/mL의 활성 보다 높은 항산화 활성을 가지고 있음을 확인하였다. 동백을 각 부위별로 추출하여 병원성균과 식중독균, 식품과 관련이 있는 세균 및 효모 등 4균주에 대하여 항미생물 활성을 검토한 결과, 동백의 수피 부위가 P. vulgaris와 B. subtilis에 대한 높은 항균성을 나타내었으며, 특히, 어린잎은 C. albicans와 T. beigelii 모두에 대하여 매우 강한 활성을 나타내었다.을 나타내었다.

번행초(Tetragonia tetragonoides)의 잎 절편으로부터 기내(in vitro) 증식을 유도 하였다. 2 mg/L BA와 0.5 mg/L NAA가 조합 처리된 MS 기본배지에서 잎 절편으로부터 직접 기관형성 과정을 통하여 부정아가 형성 되었다. 유기한 부정아는 절취하여 2 mg/L NAA와 0.5 mg/L BA이 첨가된 MS배지로 옮겨 유묘의 대량 증식을 시도하였으며, 배양 3주 후 신초의 지속적인 성장을 위해 무기염의 농도가 두배로 증가된 2MS배지로 옮겨 배양 하였다. 증식된 신초로부터 발근은 식물성장조절물질이 첨가되지 않은 1/2MS와 MS배지 모두에서 이루어 졌다.

대황(Rheum undulatum L.)의 형질전환된 뿌리의 기내(in vitro)배양으로부터 quinone계 알칼로이드 물질인 anthraquinone의 생산성을 확인하고자 하였다. 형질전환된 뿌리로부터 anthraquinone의 생산에 있어서 배지, 초기 pH, 탄소원의 농도, 광 그리고 elicitors의 영향을 조사 하였다. 형질전환된 뿌리는 6% sucrose와 0.5 mg/l GA3 그리고 50 mg/l chitosan이 첨가된 WPM 배지에 치상한 후 16/8의 광조건(16μmol m2s1)하에서 배양시 0.18 mg(생중량)의 anthraquinone의 생산성을 보여주었다. 이와같은 함량은 재배근의 약 1.3배에 해당된다.

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.