• 제목/요약/키워드: industrial biotechnology

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천궁 돌연변이 유발을 위한 최적 감마선 조사량 (Gamma-ray Irradiation on Radio Sensitivity in Cnidium officinale Makino)

  • 정진태;하보근;한종원;이정훈;이상훈;오명원;박춘근;마경호;장재기;김상훈;김진백;강시용;류재혁
    • 한국약용작물학회지
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    • 제28권5호
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    • pp.339-346
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    • 2020
  • Background: Cnidium officinale Makino have been used in traditional medicine in Northeast Asia. Although gamma-ray mutagenesis has been used to develop breeding resources with novel characteristics, research on the radiation sensitivity of C. officinale Makino is limited. Hence, the optimal gamma-ray dosage for mutation breeding in C. officinale Makino was investigated. Methods and Results: Seedstocks were exposed to doses of gamma rays (5 Gy - 50 Gy), and subsequently planted in a greenhouse. After 30 days of sowing, the survival rates and growth decreased rapidly at doses above 20 Gy, while all individuals died at 50 Gy. The median lethal dose (LD50) was 25.65 Gy, and the median reduction doses (RD50) for plant height, number of stems, and fresh weight were 12.81, 9.32, and 23.26 Gy, respectively. Post-irradiation levels of malondialdehyde (MDA), peroxidase (POD), and chlorophyll in the aerial parts of the plant were quantified using spectrophotometry. Relative to the controls, the levels of MDA and POD increased, while the level of chlorophyll decreased at doses ≥ 10 Gy, indicating cellular damage. Conclusions: A dose of 20 Gy was found to be optimal for mutation breeding in C. officinale Makino.

해양생태독성 평가용 표준시험생물로서 섬모충류 Mesodinium rubrum에 대한 연구 (Potential of Marine Ciliate Mesodinium rubrum as a Standard Test Species for Marine Ecotoxicological Study)

  • 안경호;박경수;이승민
    • 한국환경과학회지
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    • 제20권9호
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    • pp.1087-1093
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    • 2011
  • The mixotrophic marine ciliate Mesodinium rubrum possesses a highly modified algal endosymbiont as a nutrition source for the species. Accordingly, we assumed that the species can reflect the ecotoxicity on marine producer (as phytoplankton) and consumer (as zooplankton) both. A series of experiments were conducted to identify the potential of the species as a standard test species for marine ecotoxicological study. The comparison of species sensitivity on reference toxic materials was made using potassium dichromate for phytoplankton and copper chloride for zooplankton. The ciliate revealed the highest sensitivity on both reference materials among the seven test species including phytoplankton, benthic copepod and rotifer species. The toxicity end point of the species was 72hr-$EC_{50}$=1.52 mg/L (as potassium dichromate) estimated by population growth inhibition (PGI), which is more sensitive than the most sensitive phytoplankton Skeletonema costatum (72hr-$EC_{50}$=3.05 mg/L). As comparison to rotifer, it also revealed higher sensitivity on copper chloride; 72hr-$EC_{50}$=0.38 mg/L for ciliate and 48hr-$EC_{50}$=0.48 mg/L for rotifer. Also, the elutriate toxicity test of various ocean disposal wastes were conducted to identify the potential of ciliate toxicity test application using industrial waste sludges. The toxicity of leather processing waste sludge was highest on the ciliate, followed by dyeing waste sludge and dye production waste sludge as an increasing order of toxicity. 72h-$EC_{50}$ of ciliate PGI test was 1.83% and that of S. costatum 3.84% for leather waste sludge which showed highest toxicity. The toxicity test results also revealed that the highest sensitivity was observed on ciliate species on ocean disposed sludge wastes. Also, ciliate toxicity test well discriminated the degree of toxicity between sludge sources; 72h-$EC_{50}$ values were 1.83% for leather processing waste sludge, 16.75% for dye production waste sludge and 27.75% for textile production waste sludge. Even the laboratory culture methods of the species were not generally established yet, the species has high potential as the standard test species for marine toxicity test in terms of the dual reflection of phyto- and zooplankton toxicity from single test, sensitivity and test replicability.

내열성 효소를 이용한 전분으로부터 6-인산과당의 제조 (Production of Fructose 6-Phoschate from Starch Using Thermostable Enzymes)

  • 권규혁;차월석;김복희;신현재
    • KSBB Journal
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    • 제22권5호
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    • pp.345-350
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    • 2007
  • 인산당은 모든 유기체에서 발견되며 무척 다양한 유용성을 지니고 있다. 특히 glucose 1-phosphate (G1P), glucose 6-phosphate (G6P), fructose 6-phosphate (F6P) 등은 해당과 정, 당합성과정, 5탄당 인산화과정 및 캘빈회로와 같은 탄수화물 대사와 에너지 생산 대사의 주요한 핵심 중간물질이다. 특히 해당과정에서 F6P는 G6P의 이성질화반응에 의하여 생성된다. F6P의 대량생산은 전분을 이용하는 것이 가능한데, 우선 전분에 인산화효소를 가하여 G1P를 얻고, 이 G1P를 자리옮김효소 (phosphoglucomutase, GM)와 이성질화효소 (phosphoglucoisomerase, GI)를 순차적으로 적용하여 G6P와 F6P를 생산하게 된다. 효소반응의 경우 전분의 용해도 증가, 반응속도의 향상 및 미생물의 오염방지 등을 위하여 중온성 효소보다는 고온성 효소 혹은 내열성 효소가 선호된다. 본 연구는 세 가지 내열성 효소를 이용하여 전분으로부터 두 단계반응으로 F6P를 생산하는 것에 관한 것이다. 실험에 사용된 효소는 대장균에서 발현된 재조합 효소로서, 효소의 생산은 유가식 배양을 이용하였다. 1.2% 가용성 전분 200 L를 이용하여 1,253 g의 순수한 G1P를 생산하였으며 이를 이용하여 최종적으로 30% 수율로 F6P를 생산할 수 있었다. 최대수율을 얻기 위하여 반응표면분석법을 이용하여 GM : GI = 1 : 1.23, 63.5$^{\circ}C$, pH 6.85의 조건이 도출되었으며, 이 조건하에서 실험을 통하여 20 g/L의 전분을 이용하여 30% 수율로 F6P가 생성됨을 확인할 수 있었다.

Azo 염료의 분해를 위한 호알카리성 균주의 분리 및 배양조건의 최적화 (Isolation and Optimization of Cultivating Conditions of Alkalophilic Strains for Biodegradation of Azo Dye)

  • 김정목;정현채;권오진
    • KSBB Journal
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    • 제14권6호
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    • pp.718-723
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    • 1999
  • 알카리성 염색가공폐수를 처리하기 위햐여 아조염료인 Acid Red 1을 분해하는 호알카리성 균을 자연계에서 분리하였으며, 반응표면분석법인 SAS(statistical analysis system)프로그램을 이용하여 최적배양조건을 조사하였다. 염색공단 폐수처리장에서 배출되는 방류수 및 하천토양을 시료로 하여 알카리성(pH 10.0)배지에 성장하는 균 15종을 순수분리하였다. 그 중 탈색율이 가장 우수한 균주 하나를 선별하여 AR-1로 명명하였다. 탄소원(sucrose, fructose, galactose), 질소원(polypeptone, yeast extract) 및 인산원($K_2HPO_4$)이 분리균의 성장 및 탈색율에 미치는 영향을 조사한 결과, 1.0% fructose, 1.0% polypeptone, 1.0% yeast extract, 0.5% $K_2HPO_4$이 최적의 조건으로 나타났다. 반응표면분석에 의하여 염료의 생분해조건을 최적화하고자 배양온도와 배양시간에 따른 탈색율과 균성장의 특성을 모니터닝하였다. 탈색율은 34.73$^{\circ}C$에서 12.96시간, 균성장은 34.77$^{\circ}C$에서 12.97시간 배양시 각각 최적인 것으로 나타났다. 한편, 균성장과 탈색율을 다같이 만족할 수 있는 최적배양조건은 32.86~36.36$^{\circ}C$, 10.96~15.75시간으로 각각 나타났다.

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재조함 인성장호르몬의 in vitro 풀림과 재접힘 과정의 구조변화 모니터링 (Monitoring of Structural Changes during in vitro Unfolding and Refolding of Recombinant Human Growth Hormone)

  • 조태훈;채영규;안상점;이은규
    • KSBB Journal
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    • 제14권6호
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    • pp.651-654
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    • 1999
  • 재조합 인성장호르몬을 사용하여 in vitro 재접힘 공정(풀림, 희석에 의한 공기 중 산화, 그리고 투석)을 수행하였다. 표면소수성이 풀림-재접힘 공정에 중요한 역할을 한다는 것을 형광값의 변화를 통하여 알 수 있었다. 변성제의 intermediate 농도는 Urea와 Gu-HCl 경우 하나의 peak로 SDS와 Sarkosyl의 경우 두개의 peak로 나타났다. 형광값의 변화 중 특이한 점은 Urea의 경우 공기 중 산화와 투석 중의 후반부에 형광값이 증가한다는 것이다. 따라서 공기 중 산화도중 형광값이 증가하기 전에 투석을 시킨 결과 형광값이 증가를 막을 수 있었다. 아직 이 원인에 대해 자세히 알 수 없지만 계속 실험 중에 있다. 이번 실험에서 표면소수성 변화와 연관시켜 fluorescence를 이황화결합에 의한 산화된 형태를 알아보기 위한 방법으로 RP-HPLC를 마지막으로 단백질의 2차원적인 구조를 알아 보기 위해 CD를 사용하였다. CD측정 결과 Gu-HCl보다 SDS의 경우 ${\alpha}$-helices의 파괴가 더 많음을 볼 수 있었다. 재접힘된 rhGH는 본래의 2차원적 구조의 90%이상을 얻을 수 있었다. 이 실험이 기지는 의의는 이 모든 실험결과를 토대로 단백질 재접힘을 모니터링 하였다는 점이다. 즉, 형광값의 변화를 통하여 형광값이 증가하는 것은 표면 소수성이 증가함을 보이는 것으로 단백질의 풀림이 일어난 것이고 3차원적 구조가 깨지고 2차원 구조를 알아 볼 수 있는 ${\alpha}$-helices의 감소를 의미하였다. 이와는 반대로 형광값이 감소하는 것을 통해 재접힘이 일어남을 알 수 있었고, 이러한 결과를 바탕으로 단백질의 재접힘 공정의 변화과정을 형광값을 통하여 모니터링 할 수 있었다. 또한 이 실험의 목적 단백질은 rhGH이지만 다른 단백질에 적용이 될 경우 단백질 재접힘 과정을 수시로 모니터링하고 상태를 예측할 수 있으므로 산업현장에서 소량의 sample로 재접힘 상태를 쉽고 빠르게 판단할 수 있을 것이다. 단백질 재접힘 과정에서 이러한 개념의 성공적 도입은 단백질 회수 수율을 높임으로써 생물분리공정 분야의 기술 발전에 이바지 하리라 사료된다.

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Flavobacterium sp. Strain DS5에 의한 Oleic Acid로부터 산화불포화 지방산의 생산 및 분석 (Production and Analysis of Oxygenated Unsaturated Fatty Acids from Oleic Acid by Flavobacterium sp. Strain DS5)

  • 송병섭;한남수;이봉희;;김범수
    • KSBB Journal
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    • 제24권1호
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    • pp.41-46
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    • 2009
  • 식물성 오일은 다양한 생물제품 생산에 적합한 저가의 원료이다. 식물성 오일의 주성분인 oleic acid와 linoleic acid와 같은 불포화 지방산의 함량은 콩기름의 경우 각각 22%와 55%, 옥수수기름의 경우 26%와 60%, 유채기름의 경우 61%와 21%에 달한다. Hydroxy나 keto group으로 치환된 불포화 지방산들은 다른 지방산에 비해 높은 점도와 반응성 등의 특별한 화학적 특성 때문에 가소제, 계면활성제, 윤활유, 세제 등에 사용되는 유용한 산업 화합물이다. 본 연구에서는 Flavobacterium sp. strain DS5 (NRRL B-14859)를 oleic acid로부터 중간생성물로서 10-hydroxystearic acid (10-HSA)를 거쳐 10-ketostearic acid (10-KSA)로의 전환에 이용하였다. 두 개의 생물전환 생성물인 10-KSA와 10-HSA는 GC, GC-MS, $^1H-NMR$, $^{13}C-NMR$을 이용하여 정량 및 정성분석을 수행하였다. 플라스크 배양에서 10-KSA와 10-HSA의 최대 생산량은 각각 3.4 g/L와 0.5 g/L 이었다. 10-KSA 생산을 위한 최적 glucose 농도 yeast extract 농도, oleic acid 첨가시간, 첨가부피는 각각 20 g/L 이하, 5 g/L 이상, 접종 후 18 h, 0.3 ml/50 ml 이었다.

한방생균제가 한우의 체중, 혈액성상, 면역성 및 육량·육질에 미치는 영향 (Effect of Herbal Probiotic Supplementation on Weight, Blood Composition, Meat Quality and Immunity in Beef)

  • 김병기;하재정;이준구;오동엽;정대진;황은경;김수정;안용근
    • 한국식품영양학회지
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    • 제29권6호
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    • pp.860-869
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    • 2016
  • This study was a report on 60 heads of Hanwoo steers(5 treatments ${\times}$ 4 heads ${\times}$ 3 repeats). Their start weights were $361.2{\pm}14.39kg$ to $380.5{\pm}27.33kg$; 12.7 to 13.6 months of age. The aim was to investigate the effect of herbal probiotic supplementation on the weight, blood composition, meat quality and immunity in beef. In case of body weight gain, con. 1 plot(183.8 kg) in the early fattening stage, T3 plot(115.1 kg) in the middle fattening stage, and T2 plot(163.3 kg) in the late fattening stage significantly increased(p<0.05). There was a statistically significant difference in the blood content of T-Cho and IgG: con. 1 plot and T1 plot in the early fattening stage, T3 plot and T2 plots in the middle fattening stage, and con. 2 plot and T1~T3 plots in the late fattening stage had higher blood content than that of other treatment plots (p<0.05). In addition, the carcass weight was highest in the T1 plot and T2 plot among the total treatment plots(p<0.05). However, no statistical significant difference was observed in the different treatments; all individual treatment plots(T1~T3 plots) were somewhat higher than the control plots(con. 1~con. 2 plots) with respect to the carcass quantity and carcass quality. Combining and analyzing all results revealed that the dietary addition of herbal probiotics for Hanwoo steers had a positive effect on the improvement of weight, amount of meat, meat quality and immunity.

Effects of several factors on pollen germination in Platycodon grandiflorum

  • Kwon, Soo Jeong;Lee, Ui Gun;Moon, Young Ja;Cho, Gab Yeon;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.172-172
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    • 2017
  • Pollen germination and its' viability in bellflower hybrid system are of great importance. The present study was conducted to investigate the several factors underlying the pollen germination of Platycodon grandiflorum and obtain the basic data for effective artificial pollination for the production of sound specifies. The pollens of Platycodon grandiflorum started germination from one hour after planting, and the germination was actively progressed as time dependent manner. For lighting conditions, the germination of pollens under the light was faster by more than twice than that without the light. Furthermore, the germination was better in the high temperature rather than in the low temperature. The germination rate was higher in the $30^{\circ}C$. For the carbon source, the germination rate was better at the concentration of 15% regardless of the kinds. In particular, the highest value was observed with glucose. The germination rate was decreased substantially as the increasing with the higher pH. The dynamic germination of pollens was observed at the pH 5. With respect to the growth regulator, the higher concentration of NAA induced the higher the germination rate. $GA_3$ showed a good germination rate in $0.05mg{\cdot}L^{-1}$. Meanwhile, for kinetin, lower concentration increased the germination rate, unlike NAA. The higher concentrations of boric acid degraded the germination rate, and the addition of boric acid of $10mg{\cdot}L^{-1}$ demonstrated higher germination rate than the addition of other growth regulators. Notably, the addition of asparaginic acid exhibited the similar results in all test sectors regardless of concentration, whereas a little higher result was observed in the high concentration sector. Taken together, the results concluded that the boric acid was considered as one of the essential minerals that played an important role on the germination of pollens.

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Effects of various medium on mass propagation of in vitro cultured Platycodon grandiflorum with yellow green petals

  • Kwon, Soo Jeong;Han, Eun Ji;Moon, Young Ja;Cho, Gab Yeon;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.171-171
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    • 2017
  • Propagation by crown division in Platycodon grandiflorum is too slow for producing many plants in a short time although the plants are uniform. This study was performed to enhance the mass propagation for Platycodon grandiflorum containing yellow green petals via various medium compositions and the growth regulators. The nodes containing yellow green petals were used as materials to execute the study with a variety of MS medium concentrations. The 1/4MS medium showed the best development of adventitious root, while the 1/2MS medium exhibited the potential growth. The higher the concentration of sucrose showed the better development and growth of both shoots and adventitious roots. Many adventitious roots were developed at the controlled culture medium at pH 4.8 with a tendency of suppression with higher levels of pH. Also, the cultivated node and leaf explants with the treatments of simple and combined applications with auxin and cytokinin at the 1/4MS culture medium with adding 5% of sucrose were used to identify the influences of growth regulators. The regeneration of plantlets at the 234single application showed a good result with the addition of BA $1mg{\cdot}L^{-1}$ and the development and growth of adventitious roots appeared to be good at the addition of NAA $1mg{\cdot}L^{-1}$. For the combined applications, the regeneration of plantlets and the development of adventitious roots were prosperous at the combined applications with BA $0.1mg{\cdot}L^{-1}$ and IAA $0.5mg{\cdot}L^{-1}$. The addition of IAA for the leaf explants induced a number of plantlets that showed the potential regeneration, and the highest results was obtained from the combined applications of both BA $1mg{\cdot}L^{-1}$ and IAA $2.5mg{\cdot}L^{-1}$. In addition, the development of adventitious roots showed the satisfactory results at the combined application of both BA $1mg{\cdot}L^{-1}$ and IAA $0.5mg{\cdot}L^{-1}$.

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Comparative physiological and proteomic analysis of leaf in response to cadmium stress in sorghum

  • Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Kim, Sang-Woo;Lee, Moon-Soon;Chung, Keun-Yook;Woo, Sun-Hee
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.124-124
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    • 2017
  • Cadmium (Cd) is of particular concern because of its widespread occurrence and high toxicity and may cause serious morpho-physiological and molecular abnormalities in in plants. The present study was performed to explore Cd-induced morpho-physiological alterations and their potentiality associated mechanisms in Sorghum bicolor leaves at the protein level. Ten-day-old sorghum seedlings were exposed to different concentrations (0, 100, and $150{\mu}M$) of $CdCl_2$, and different morpho-physiological responses were recorded. The effects of Cd exposure on protein expression patterns in S. bicolor were investigated using two-dimensional gel electrophoresis (2-DE) in samples derived from the leaves of both control and Cd-treated seedlings. The observed morphological changes revealed that the plants treated with Cd displayed dramatically altered shoot lengths, fresh weights, and relative water content. In addition, the concentration of Cd was markedly increased by treatment with Cd, and the amount of Cd taken up by the shoots was significantly and directly correlated with the applied level of Cd. Using the 2-DE method, a total of 33 differentially expressed protein spots were analyzed using MALDI-TOF/TOF MS. Of these, treatment with Cd resulted in significant increases in 15 proteins and decreases in 18 proteins. Significant changes were absorbed in the levels of proteins known to be involved in carbohydrate metabolism, transcriptional regulation, translation and stress responses. Proteomic results revealed that Cd stress had an inhibitory effect on carbon fixation, ATP production and the regulation of protein synthesis. In addition, the up-regulation of glutathione S-transferase and cytochrome P450 may play a significant role in Cd-related toxicity and stress responses. Our study provides insights into the integrated molecular mechanisms involved in response to Cd and the effects of Cd on the growth and physiological characteristics of sorghum seedlings. The upregulation of these stress-related genes may be candidates for further research and use in genetic manipulation of sorghum tolerance to Cd stress.

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