• Parasites, Hosts and Diseases
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• 제51권2호
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• pp.177-182
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• 2013

The purpose of the present study was to evaluate the potential role of the 27-Kilodalton (KDa) antigen versus Fasciola gigantica adult worm regurge antigens in a DOT-Blot assay and to assess this assay as a practical tool for diagnosis fascioliasis in Egyptian patients. Fasciola gigantica antigen of an approximate molecular mass 27- (KDa) was obtained from adult worms by a simple elution SDS-PAGE. A Dot-Blot was developed comparatively to adult worm regurge antigens for the detection of specific antibodies from patients infected with F. gigantica in Egypt. Control sera were obtained from patients with other parasitic infections and healthy volunteers to assess the test and compare between the antigens. The sensitivity, specificity, positive and negative predictive values of Dot-Blot using the adult worm regurge were 80%, 90%, 94.1%, and 69.2% respectively, while those using 27-KDa were 100% which confirms the diagnostic potential of this antigen. All patients infected with Fasciola were positive, with cross reactivity reported with Schistosoma mansoni serum samples. This 27-KDa Dot-Blot assay showed to be a promising test which can be used for serodiagnosis of fascioliasis in Egyptian patients especially, those presenting with hepatic disease. It is specific, sensitive and easy to perform method for the rapid diagnosis particularly when more complex laboratory tests are unavailable.

• 미생물학회지
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• 제37권4호
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• pp.273-276
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• 2001

본 연구에서는 중금속, 특히 은(銀)을 축적하는 세균을 지하수로부터 분리, 동정하고, 분리 균주의 특성 및 중금속 축적조건을 규명하여 최대축적이 일어날 수 있는 조건을 검토하였다. 서울시 지하수 10개 정점으로부터 분리된 은(銀) 축적 세균 중 가장 축적능이 뛰어난 두 균주를 선별하였으며 이들은 Pseudomonas fluorescens와 Bacillus cereus로 동정되었다. 본 분리 균주는 은(銀)의 농도가 5 ppm, 10 ppm, 20 ppm으로 높아질수록 생장 저해를 심하게 받았다. pH가 낮을수록 세포의 성장이 저하되기는 하였지만 세포 당 은(銀) 축적율은 오히려 증가하였으며 24시간 이내에 최대축적이 이루어졌다. 이때 은(銀) 축적량은 Pseudomonas fluorescens의 경우 약 1.9 $mg/g{\cdot}cell$ 이었으며 Bacillus cereus는 약 1.65 $mg/g{\cdot}cell$ 이었다. 은(銀) 처리시, Bacillus cereus에서 126 KDa, 89 KDa, 25 KDa 등 3개의 단백질이 특이적으로 증가하였으며, Pseudomonas fluorescens에서는 101 KDa, 68 KDa, 27 KDa 크기의 단백질이 감소하였으나 34 KDa 크기의 새로운 단백질이 유도됨을 확인하였다.

• 한국균학회지
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• 제27권2호통권89호
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• pp.100-106
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• 1999

Agrocybe cylindracea의 배양방법별 균사체 및 다당류 생산수율비교 실험에서는 모두 진탕배양이 효과적이었다. 다당류의 정제는 조단백다당류를 이용하여 DEAE-cellulose column에 의한 1차 정제를 행하였고, 최종으로 Sepharose 2B를 이용한 2차 정제를 실시하여 최종적으로 균사체내다당류(ACIPDG, ACIPAG)와 균사체외다당류(ACEPDG, ACEPAG)를 얻었다. ACEPDG는 총당 75.8%, ACEPAG는 총당 65.4%를 함유하였으며, ACIPDG는 총당 89.2%, ACIPAG는 총당 54.2%의 조성을 보였다. 다당류 분획물들의 분자량을 측정한 결과, 모두 10만이 넘는 거대분자로 ACEPDG 300KDa에서 ACIPAG 600KDa까지 나타났다. 다당류의 조성을 HPLC로 분석한 결과, ACEPDG분획물은 glucose, inositol이 검출되었으며 ACIPDG, ACEPAG, ACIPAG 분획물은 glucose, fructose, inositol이 검출되었다.

• 한국수산과학회지
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• 제27권5호
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• pp.482-494
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• 1994

우리나라에서 대량 생산되고 있는 진주담치 근육에서 Triton X-100를 사용하여 carotenoprotein을 추출, 정제하여 그 특성을 실험한 결과를 요약하면 다음과 같다. Carotenoprotein(${\lambda}_{max}=462nm$)를 $20\%$(w/v) $(NH_4)_2SO_4$, DEAE-cellulose 이온교환수지 그리고 Sephacryl S-300 겔 크로마토그래피를 통해 정제한 결과, 정제도는 5배 증가하였고 수율은 $1.85\%$였으며, carotenoprotein의 분자량은 약 372KDa(gel filtration)이었고, SDS-PAG 전기영동으로 단백질의 subunit를 확인한 결과, 60KDa($23.70\%$), 46.9KDa($9.14\%$), 26KDa($49.14\%$) 및 13KDa($18.02\%$)로 이루어져 있었다. Carotenoprotein의 안정성은 pH 4.0 이하의 강산 및 pH 10.0 이상의 강알칼리에서는 단백질이 불안정하였고, SDS 변성제를 $0.2\%$(w/v)되게 첨가하여 녹인 후 가시부 영역($400{\sim}600nm$)에서 최대파장 변화를 보면 462nm에서 456nm로 약 6nm 단파장 영역으로 이동한 것을 알 수 있었다. Carotenoprotein의 구성아미노산 조성은 threonine이 $35.90\%$로 다른 아미노산에 비해 함량이 높았으며, 필수아미노산과 소수성 아미노산의 함량은 전체아미노산 함량의 각각 $62.76\%$와 $25.02\%$를 차지하였고, methionine, histidine 및 proline은 존재하지 않았다. Carotenoprotein 1g당 총지질의 함량은 256mg였고, 그 구성지방산의 조성은 polyene산이 $62.4\%$로 가장 많았고 다음으로 포화산($19.58\%$)및 monoene산($18\%$) 순이었다. 특히, 16:4n-3의 조성비가 $33.90\%$로 상당히 높게 나타났고, EPA, DHA 등 w3 계의 장쇄(長鎖)고도불포화지방산의 조성비가 $22.13\%$로 높게 나타났으며, 양적으로 많은 지방산은 포화산 중에는 palmitic acid($C_{16:0}$), monoene산 중에서는 palmitoleic acid($C_{16:1}$), polyene산 중에서는 hexadecatetraenoic acid($C_{16:4}$), eicosapentaenoic acid ($C_{20:5}$), docosahexaenoic acid($C_{22:6}$) 등이 함량이 많았다. Carotenoprotein으로부터 분리한 총 caroteoid 함량은 $8.80mg\%$였고, 주 carotenoid는 mytiloxanthin($74.79\%$), 3,4,3'-trihydroxy-7',8'-didehydro-${\beta}$-carotene($18.26\%$)였다.

• Mycobiology
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• 제37권2호
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• pp.121-127
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• 2009

Purification and characterization of intracellular cellulase produced by A. oryzae ITCC-4857.01 are reported. The enzyme was purified by ion-exchange chromatography using DEAE-cellulose followed by Gel filtration. The purification achieved was 41 fold from the crude extract with yield of 27%. The purified enzyme showed single band on poly acrylamide gel. The molecular weight as determined by SDS-PAGE and gel filtration was 38 KDa and 38.6 KDa respectively and contained only one subunit. The enzyme is glycoprotien as nature and contained 0.67% neutral sugar. The apparent Km value of the enzyme against cellulose was 0.83%. The enzyme showed the highest relative ativities on CMC followed by avicel, salicin and filter paper. The optimum pH of activity was 5.5 and very slight activity was observed at or above pH 7.5 as well as bellow pH 3.5. The optimum tempreture of the activity was $45^{\circ}C$ and the highest activity was exhibited in 35 to $45^{\circ}C$. The enzyme lost their activities almost completely (95${\sim}$100%) at $80^{\circ}C$ or above and as well as bellow $25^{\circ}C$.

• Animal Bioscience
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• 제34권4호
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• pp.724-733
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• 2021

Objective: The objectives of this study were to investigate the direct antioxidative effect of 90 Kda heat shock protein (Hsp90) obtained from duck muscle. Methods: The interaction of Hsp90 with phospholipids and oxidized phospholipids was studied with surface plasmon resonance (SPR), and their further oxidation in the presence of Hsp90 was evaluated with thiobarbituric acid reactive substances (TBARS) assay. The scavenging effect on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonic acid (ABTS) was measured, and the electron paramagnetic resonance (EPR) spectroscopy in combination with 5-tert-Butoxycarbonyl-5-methyl-1-pyrroline-N-oxide and 2-phenyl-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) was utilized to determine the abilities of Hsp90 in scavenging hydroxyl and PTIO radicals. Results: SPR showed Hsp90 could bind with both phospholipids and oxidized phospholipids, and prevent their further oxidation by the TBARS assay. The DPPH and ABTS scavenging activity increased with Hsp90 concentration, and could reach 27% and 20% respectively at the protein concentration of 50 μM. The EPR spectra demonstrated Hsp90 could directly scavenge ·OH and PTIO· radicals. Conclusion: This suggests that Hsp90, a natural antioxidant in meat, may play an important role in cellular defense against oxidative stress, and may have potential use in meat products.

• 핵의학기술
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• 제27권2호
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• pp.133-136
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• 2023

Purpose: Prolactin in the blood is separated into three types, and over 90% of prolactin presents as a double monomer (23 KDa). Rarely, it can exist in the size of big prolactin (150 KDa), which is called macroprolactin and is known as an autoantibody complex. When macroprolactin accounts for more than 60% of prolactin in the blood, it is called macroprolactinemia. The presence of such macroprolactin was first reported in a patient with hyperprolactinemia but without typical symptoms. Macroprolactinemia is emerging as an important cause of idiopathic hyperprolactinemia. The polyethylene glycol (PEG) precipitation method using the property of precipitating large-molecular-weight proteins is simple and recently has been widely used as a screening test. The results are in good agreement with the results of gel chromatography. The purpose of this study was to confirm the measurement method and reference value verification of monomeric prolactin in blood prolactin using the PEG precipitation method. Materials and Methods: For 40 examinees who visited the Gangnam Center of Seoul National University Hospital in 2021, the prolactin level was verified using radioimmunoassay (RIA). For macroprolactinemia PEG precipitation method, 25% PEG (molecular weight 6000kDa) solution and serum were mixed in equal amounts in a test tube, then left at room temperature for 20 minutes and centrifuged at 4℃ for 30 minutes (1500g). The prolactin level was measured in the supernatant. Results : After confirming that more than 90% of the 40 tested samples within the reference range <25 ng/mL, the same value as the reference value for prolactin was applied. Since the concentration of monomeric prolactin in serum from which macroprolactin has been removed from blood is diluted 1:1 with PEG, our laboratory is currently reporting the result by multiplying the result by a dilution factor of 2. Conclusion: Radioimmunoassay using PEG precipitation method using the property of precipitating large molecular weight proteins is simple and effective for quantitative measurement of monomeric prolactin in blood prolactin.

• 농업생명과학연구
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• 제43권1호
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• pp.25-33
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• 2009

본 연구는 홀스타인, 한국재래산양 및 한우유로부터 glycomacropeptide(GMP)를 분리하였으며, 각 GMP의 trypsin 가수분해물의 혈소판응집 억제 효과를 in vitro상에서 알아보았다. 홀스타인, 한국재래산양 및 한우의 GMP는 분자량이 모두 약 20 KDa이었으며, sialic acid 함량은 각각 $36.86{\pm}2.36$, $37.98{\pm}1.27$ 및 $31.19{\pm}1.87{\mu}g/mg$이었다. 또한 모든 개체의 GMP에서 tyrosine이 검출되었다. 홀스타인, 한국재래산양 및 한우 GMP의 trypsin 가수분해물에 의한 혈소판 응집 억제율은 반응 30초에 4.02, 5.51 및 12.77%로 각각 나타나 시간이 경과할수록 감소하는 경향을 보였다. 혈소판의 현미경 관찰 결과 가수분해물 첨가 후 혈소판 수가 증가하였으나, 첨가 후 30초가 경과한 시점부터 혈소판 수가 감소하기 시작하여 120초 후에는 관찰 할 수 없었다. 본 실험 결과 bovine 및 caprine GMP의 trypsin 가수분해물에서 혈소판 응집을 억제할 수 있는 small peptide가 있는 것으로 생각되며, 향후 이러한 연구는 심근경색증 및 뇌혈전증을 예방할 수 있는 생리활성 물질로 이용될 수 있을 것이라 생각된다.

• 한국식품과학회지
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• 제27권6호
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• pp.860-865
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• 1995

처리 방법을 달리한 메밀(날메밀, 볶은메밀, 찐메밀)에서 효소적 방법으로 비소화성 다당과 가용성 다당을 분리하였다. 가용성 다당은 저분자 가용성 다당과 고분자 가용성 다당으로 비소화성 다당은 crude hemicellulose(CH)와 alcohol insoluble hemicellulose(AIH), residue 그리고 저분자 불용성 다당으로 각각 분리하였다. 다당의 함량은 조리방법에 따라 차이를 나타냈으며 가용성 다당 중 고분자 가용성 다당의 함량은 $5.5{\sim}9.4%$였으며, 저분자 가용성 다당의 함량은 $75.9{\sim}84.2%$였다. 비소화성 다당 중 CH는 $3.2{\sim}9.6%$, AIH는 $0.9{\sim}1.7%$, residue는 $2.0{\sim}2.4%$, 저분자 불용성 다당은 $0.8{\sim}4.2%$였다. 가용성 다당에서는 유리당이 검출되었으나 비소화성 다당에서는 유리당이 검출되지 않았으며 모든 다당 분획에서 단백질이 검출되었다. 가용성 다당의 주요 성분당은 glucose였으며, 비소화성 다당에서는 glucose 이외에 rhamnose, arabinose, xylose, galactose, mannose가 확인되었다.

• KSBB Journal
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• 제27권3호
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• pp.177-185
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• 2012

Inonotus obliquus mushroom, which is a fungus belonging to Hymenochaetaceae family, is known to grow on birth trees in colder northern climates and to be a fungal parasite that draws nutrients out of living trees rather than from the ground. For the separation of protein-bound polysaccharide (PBP) from the culture broth and mycelium of Inonotus obliquus, three well known extraction methods namely hot water, ultrasound and microwave were used. The best extraction conditions to separate the PBP (64.94 mg/g) from mycelium by microwave were found to be for 1 hour and $150^{\circ}C$. The possibility for concentration of extracted PBP solution by using membrane was also studied. The extracted PBP solution was concentrated effectively by using an ultrafiltration membrane and the molecular weight cut off (MWCO) is 30 KDa. It was observed that a concentration by the ultrafiltration membrane is essential not only for the development of clean separation technology but also for enhanced production of PBP. As a result, we have shown that PBP in the final concentrated solution showed approximately 10 times higher than that in the crude solution by application of the developed separation systems. The separation yield of PBP was about 89.79% by gel filtration of purification steps and the purified product was confirmed to be PBP by using FT-IR.