• Title/Summary/Keyword: biochemical activity

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파밤나방(Spodoptera exigua (Hubner))의 살충제 감수성 변이-에스테라제와 아세틸콜린에스테라제 활력 (Variation in Insecticide Susceptibilities of the Beet Armyworm, Spodoptera exigua (Hubner): Esterase and Acetylcholinesterase Activities)

  • 김용균;이준익;강성영;한상찬
    • 한국응용곤충학회지
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    • 제36권2호
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    • pp.172-178
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    • 1997
  • 파밤나방(Spodoptera exigua (Hubner))은 야외집단과 실내집단들간에 살충제 감수성에서 큰 변이를 보였다. 조사된 모든 약제에서 야외집단은 비선발 실내집단보다 높은 약제 저항능력을 보였다. 약제 선발된 실내집단은 선발 약제에 대해 저항능력을 높게 발현시켰다. 이러한 약제 저항능력의 변이가 아세틸콜린에스테라제와 에스테라제 활력 변이와 비교 분석되었다. 약제 저항능력이 높은 집단일수록 아세틸콜린에스테라제 활력이 낮아지고 에스테라제활력은 높아졌다. 아세틸콜린에스테라제의 Km(Michaelis-Menten constant)값은 약제저항능력이 높은 집단일수록 증가하는 것으로 기질에 대한 이 효소의 친화도가 낮아졌다. 이 효소의 효소활력 변이는 같은 집단일수록 증가하는 것으로 기질에 대한 이 효소의 친화도가 낮아졌다. 이 효소의 효소활력 변이는 같은 집단 내에서 발육단계에 따라 차이를 나타내어 2, 3, 4령충보다는 5령충에서 특히 Km값이 증가하였다. 파밤나방의 에스테라제는 6.5% 비변성조건의 겔에서 21개 밴드를 보였다. 이중 E2, E7, E8, E11, E16, E7 밴드들은 약제저항능력이 높은 집단일수록 발현 빈도수가 높았다. 이상의 결과는 파밤나방의 살충제 감수성변이에 해독효소활력증가와 작용점변환이 포함됨을 나타낸다.

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The effect of biomechanical stimulation on osteoblast differentiation of human jaw periosteum-derived stem cells

  • Lee, Ju-Min;Kim, Min-Gu;Byun, June-Ho;Kim, Gyoo-Cheon;Ro, Jung-Hoon;Hwang, Dae-Seok;Choi, Byul-Bora;Park, Geun-Chul;Kim, Uk-Kyu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제39권
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    • pp.7.1-7.9
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    • 2017
  • Background: This study was to investigate the effect of biomechanical stimulation on osteoblast differentiation of human periosteal-derived stem cell using the newly developed bioreactor. Methods: Human periosteal-derived stem cells were harvested from the mandible during the extraction of an impacted third molar. Using the new bioreactor, 4% cyclic equibiaxial tension force (0.5 Hz) was applied for 2 and 8 h on the stem cells and cultured for 3, 7, and 14 days on the osteogenic medium. Biochemical changes of the osteoblasts after the biomechanical stimulation were investigated. No treatment group was referred to as control group. Results: Alkaline phosphatase (ALP) activity and ALP messenger RNA (mRNA) expression level were higher in the strain group than those in the control group. The osteocalcin and osteonectin mRNA expressions were higher in the strain group compared to those in the control group on days 7 and 14. The vascular endothelial growth factor (VEGF) mRNA expression was higher in the strain group in comparison to that in the control group. Concentration of alizarin red S corresponding to calcium content was higher in the strain group than in the control group. Conclusions: The study suggests that cyclic tension force could influence the osteoblast differentiation of periosteal-derived stem cells under optimal stimulation condition and the force could be applicable for tissue engineering.

간에서의 RNA, 단백질 생합성에 미치는 인삼성분의 생화학적 연구 (Biochemical Studies of Ginseng Saponin on RNA and Protein Biosynthesis in the Rat Liver)

  • Oura Hikokichi
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1988년도 학술대회지
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    • pp.1-10
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    • 1988
  • 조사포닌을 복강내 투여하면 1) 핵내의 RNA polymerase의 활성도, 2) 핵내의 RNA합성, 3) 세포질의 RNA합성,4) 세포질내의 폴리리보좀 함량, 5) in vitro 상태에서의 쥐간의 polysome과 micro-some으로의 아미노산 유입율, 6) 방사능 표지된 아미노산의 혈청 단백질로의 유입율이 증가하였음을 과거에 보고 한 바 있으며 또한 4주간 조사포닌을 투여한 쥐에서 적출한 간세포를 전자 현미경으로 조사한 결과, 조면 소포체가 상당히 증가하였으며 초원심분리기로서 막에 결합한 ribosome에서의 polysome함량의 증가를 확인하였다. 최근 streptozotocin으로 유도한 단백질 결핍성 당뇨병 쥐에 $Rb_2$를 계속적으로 주사한 결과 blood urea nitrogen과 간내의 urea 농도가 현저히 감소하였으며 혈청내의 총단백질과 알부민의 농도가 대조군의 수치에 비하여 증가한 반면 간내의 RNA와 총 ribosome, 막에 결합된 ribosome의 함량이 증가하였다. 또한 $Rb_2$투여로 혈청내의 총단백질로의 방사능 표지 전구물질의 유입량이 증가하였으며 당뇨쥐에서의 질소균형을 개선시켰다. 이러한 실험적 결과에 근거하여 인삼 사포닌은 대사를 촉진시키고 RNA, 단백질 합성 등을 촉진하는 것으로 생각된다.

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모 공단지역 사무직 근로자들의 요중 N-acetyl-$\beta$-D-glucosaminidase 역가에 관한 연구 (A Study on Urinary N-acetyl-$\beta$-D-glucosaminidase Activities of Office Workers in a Certain Industrial Complex Area)

  • 김화성;리갑수;이성수;안규동;이병국
    • Journal of Preventive Medicine and Public Health
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    • 제27권3호
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    • pp.547-556
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    • 1994
  • In order to identify the necessary information of biochemical Indices for renal effect of lead for the early detection in medical surveillance of lead worker, the reference values of urinary N-acetyl-$\beta$-D-glucosaminidase (NAG) activities were studied with 205 office workers in one industrial complex area who were not exposed to lead occupationally. While study variables selected for lead exposure were blood lead (PbB), blood zinc protoporphyrin(ZPP) and $\delta$-aminolevulinic acid (DALA) in urine, those for renal effect were urinary N-acetyl-$\beta$-D-glucosaminidase (NAG), blood urea nitrogen (BUN), serum creatinine(Cr), serum uric acid (Ua), and urinary total protein(U-TP). The results obtained were as follows: 1. The mean values of blood lead, ZPP and DALA in all subjects were $14.39{\pm}4.02{\mu}g/dl,\;21.61{\pm}8.00{\mu}g/dl,\;and\;2.73{\pm}0.90mg/l$ respectively. 2. The mean value of urinary NAG activities in all subjects was $3.51{\pm}2.01U/l$. The mean value of urinary NAG activities, which calculated from NAG activities divided by urinary creatinine concentration (CNAG), was $5.42{\pm}5.53U/g$ creatinine and log-arithmic normal distributed. 3. The reference value of urinary NAG activity was 12.06 U/g creatinine(95% CU=10.57-14.76 U/g creatinine). 4. Logarithmic CNAG(r=0.781 p<0.0l), U-TP(r=0.670 p<0.01) and ZPP(r=0.172 p<0.05) showed statistically significant correlation with CNAG.

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Biligrafin 투여 마우스 간세포의 미세구조적 및 세포화학적 연구 (Some Observations on the Organelles Participating in the Biliary Excretion in the Hepatocyte of the Biligrafin Injected Mouse)

  • 김향;신영철
    • Applied Microscopy
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    • 제23권2호
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    • pp.53-77
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    • 1993
  • In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.

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Cellulase 및 Xylanase를 분비하는 Bacillus licheniformis DK42의 분리 및 효소 특성 (Isolation of a Bacillus licheniformis DK42 Producing Cellulase and Xylanase, and Properties of the Enzymes)

  • 김민정;임수진;강대경
    • Journal of Animal Science and Technology
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    • 제50권3호
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    • pp.429-436
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    • 2008
  • 사료곡물의 효율적 이용을 목적으로 cellulase 및 xylanase를 분비하는 미생물을 분리한 후 가장 생산성이 높은 균주를 선발하였다. 선발된 DK42 균주의 형태, 당 이용성 및 16S rRNA 유전자 서열분석 등을 통해 동정한 결과, Bacillus licheniformis에 속하는 것으로 나타났으므로 선발한 균주를 B. licheniformis DK42로 명명하였다. B. licheniformis DK42가 분비하는 cellulase 효소 활성은 대수생장기 중반부터 급격히 증가하였고, 균의 생장이 정체기에 이르면 효소의 활성도 더 이상 증가하지 않는 것으로 나타났다. 한편, xylanase 효소 활성은 세포 생장과 더불어 대수생장기 초기부터 지속적으로 증가하였으며, 대수생장기 후반에 최대 활성을 나타내고 효소 활성이 더 이상 증가하지 않는 것으로 나타났다. Cellulase 활성의 경우에는 최적 온도가 45℃이었고 10℃의 저온에서도 최대 활성의 50% 정도의 활성을 나타내었으며, xylanase는 cellulase 보다 약간 높은 55℃에서 최고 활성을 나타내었다. 한편, 두 효소의 열안정성을 조사한 결과, cellulase는 45℃에서는 2시간 후에도 효소의 활성이 안정하게 유지되었으나, xylanase는 45℃에서도 약간 이상의 온도에 방치한 경우에는 시간의 경과에 따라 효소의 활성이 점진적으로 감소하였다. 두 효소의 최적 pH를 조사한 결과, 두 효소 모두 pH 6.0에서 최대의 효소 활성을 나타내었으며, cellulase 활성은 pH 5.0부터 pH 8.0까지 상대적으로 넓은 범위에서 활성을 유지한 반면, xylanase 활성의 경우에는 pH 5.0 이하 또는 pH 8.0 이상에서는 활성이 급격히 저하되었다. Ⅴ. 사 사이 연구는 2006년도 단국대학교 대학연구비의 지원으로 연구된 것으로 이에 감사드립니다

Bacillus subtilis를 이용한 대두 발효식품의 혈전용해능

  • 정영기
    • 한국생명과학회:학술대회논문집
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    • 한국생명과학회 2001년도 제32회 학술심포지움
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    • pp.67-86
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    • 2001
  • A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

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Rice OsACDR1 (Oryza sativa Accelerated Cell Death and Resistance 1) Is a Potential Positive Regulator of Fungal Disease Resistance

  • Kim, Jung-A;Cho, Kyoungwon;Singh, Raksha;Jung, Young-Ho;Jeong, Seung-Hee;Kim, So-Hee;Lee, Jae-eun;Cho, Yoon-Seong;Agrawal, Ganesh K.;Rakwal, Randeep;Tamogami, Shigeru;Kersten, Birgit;Jeon, Jong-Seong;An, Gynheung;Jwa, Nam-Soo
    • Molecules and Cells
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    • 제28권5호
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    • pp.431-439
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    • 2009
  • Rice Oryza sativa accelerated cell death and resistance 1 (OsACDR1) encodes a putative Raf-like mitogen-activated protein kinase kinase kinase (MAPKKK). We had previously reported upregulation of the OsACDR1 transcript by a range of environmental stimuli involved in eliciting defense-related pathways. Here we apply biochemical, gain and loss-of-function approaches to characterize OsACDR1 function in rice. The OsACDR1 protein showed autophosphorylation and possessed kinase activity. Rice plants overexpressing OsACDR1 exhibited spontaneous hypersensitive response (HR)-like lesions on leaves, upregulation of defense-related marker genes and accumulation of phenolic compounds and secondary metabolites (phytoalexins). These transgenic plants also acquired enhanced resistance to a fungal pathogen (Magnaporthe grisea) and showed inhibition of appressorial penetration on the leaf surface. In contrast, loss-of-function and RNA silenced OsACDR1 rice mutant plants showed downregulation of defense-related marker genes expressions and susceptibility to M. grisea. Furthermore, transient expression of an OsACDR1:GFP fusion protein in rice protoplast and onion epidermal cells revealed its localization to the nucleus. These results indicate that OsACDR1 plays an important role in the positive regulation of disease resistance in rice.

참깨의 볶음 조건이 참깨 착즙액의 이화학적 및 생화학적 특성에 미치는 영향 (Studies on the physicochemical and biochemical characteristics in sesame seed juice under different roasting conditions)

  • 박혜정;김지윤;박성환;이상현;장정수;이문현
    • 한국식품과학회지
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    • 제49권4호
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    • pp.421-429
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    • 2017
  • 본 연구에서는 참깨 착즙액의 볶음 온도가 영양 성분, 소화효소 활성 및 항산화 활성에 미치는 영향을 조사하였다. 참깨를 160, 200, $240^{\circ}C$에서 3분간 볶은 뒤 저속 녹즙기로 착즙하였다. 짧은 시간의 볶음 처리로 인해 모든 참깨 착즙액에서 벤조피렌 및 트랜스지방산은 검출되지 않거나 극소량만 검출되었다. 참깨 착즙액은 무기질, 비타민 및 지방산과 같은 다량의 무기질을 함유하며, 볶음 시간이 증가함에 따라 무기질, 비타민 B1, B3 및 세사몰 함량은 증가되는 것을 확인하였다; 하지만 지방산, 비타민 $B_2$, 세사민 및 세사몰린은 감소되었다. 참깨 착즙액의 항산화 성분 및 항산화 활성은 볶음 온도가 증가함에 따라 증가되었고, $240^{\circ}C$에서 가장 높은 항산화 활성을 나타냈다. 그러므로 본 결과는 항산화 활성이 높은 볶은 참깨 착즙액이 체내 산화적 손상을 예방하는데 도움이 될 것으로 제안한다.

Bacillus cereus에 대한 길항적 저해 작용과 biogenic amines 분해 능력을 지닌 Bacillus licheniformis SCK A08 균의 특성 (Characterization of Bacillus licheniformis SCK A08 with Antagonistic Property Against Bacillus cereus and Degrading Capacity of Biogenic Amines)

  • 이은실;김용상;류명선;정도연;엄태붕;조성호
    • 한국식품위생안전성학회지
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    • 제29권1호
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    • pp.40-46
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    • 2014
  • Biogenic amine은 주로 단백질이 풍부한 전통장류 중 된장과 간장에서 높게 발생하고 있다. 현재까지 국내외 식중독 사고를 보면 biogenic amine중 tyramine, histamine과 putrescine에 의한 보고가 있다. 전통장류의 biogenic amine에 대한 위생적인 문제를 해결하기 위해 전통장류 제조에 적합한 발효미생물 중에서 biogenic amine 비생성 및 분해하는 미생물을 선발하여 종균으로 적용한다면 biogenic amine에 대한 문제를 해결할 수 있을 것이다. 이러한 문제의 해결을 위해 본 연구에서는 우수한 발효능력과 함께 유해균에 대해 강한 길항 능력을 지니며, biogenic amine을 생산하지 않지만 높은 분해능력을 보이는 균주를 선발하여 장류제조 종균으로의 이용가능성을 실증하였다. 결론적으로 선발된 장류발효균주를 종균으로 이용하여 메주제조 및 청국장 제조 등에 이용한다면 biogenic amine과 Bacillus cereus를 안전한 수준으로 낮출 수 있을 것으로 사료되었고, 전통장류의 HACCP적용을 위한 HAZARD를 관리하는 우수한 수단으로 이용될 수 있을 것이다. 또한 향후에는 단일 균종만 종균으로 사용할 경우 전통장류의 우수한 풍미를 재현하는데 한계가 있기 때문에 종균들을 적절하게 조합한 혼합발효를 통해 제조한다면 풍미가 좋을 뿐만 아니라, 위생문제를 해결한 우수한 장류를 생산할 수 있을 것으로 판단되었다.