• 한국식품과학회지
• /
• 제20권1호
• /
• pp.34-39
• /
• 1988

여러가지 육가공품(肉加工品)중 특정 단백질 원료의 첨가여부를 판정하여 변조식품(變造食品)을 검출하는 한 방법으로서, 각종 육류단백질, non-meat protein, 어육(魚肉)가공품을 대상으로 disc SDS-Poly acrylamide gel electrophoresis의 사용 가능성을 실험하였다. Total protein fraction에 대한 전기영동 결과 복잡하고 많은 band를 보여 각 시료에 고유한 특성을 찾아보기 어려웠다. Low salt-soluble protein fraction에서는 total protein fraction 에서 보다 band 수가 상당히 감소함을 보였고 각 단백질 원료에 대하여 보다 고유한 band pattern을 나타내었다. Acetone-insoluble protein fraction 에서는 non-meat protein의 경우 육류단백질과 상당히 다른 경향을 나타내었고. 소세지 원료의 가열처리에 의하여 단백질의 band수와 양이 감소하였다. 따라서 적당한 단백질 추출조건(抽出條件)을 설정하여 전기영동을 실시하면, 특정(特定) 단백질을 첨가한 변조식품의 검출이 가능해질 것으로 생각된다.

• Toxicological Research
• /
• 제17권
• /
• pp.251-256
• /
• 2001

The expression of phase II detoxifying enzymes is affected by a variety of compounds and the induction of the enzymes plays an essential role in chemoprevention. A variety of phytochemicals such as sulfur-containing chemoprotective agents (SCC) may trigger cellular signals and activate phase II gene expression through ARE activation. see induces glutathione S-transferases. Studies were conducted to investigate the role of mitogen-activated protein (MAP) kinase and phosphatidylinositol 3-kinase (PI3-kinase) in the induction of GST (e.g. rGSTA2) by sec. We also studied the MAP kinase pathway responsible for the GST expression by see and compared that with the pathway activated by oxidative stress as a result of sulfur amino acids deprivation (SAAD). see inhibited phosphorylation of ERK1/2 although the effect of see on JNK and p38 MAP kinase was minimal. Wortmannin and LY294002. PI3-kinase inhibitors. abolished the increases in rGSTA2 mRNA and protein levels by SCC. Deprivation of cystine and methionine caused oxidative stress in H4IIE cells. as evidenced by a decrease in the reduced glutathione and an increase in prooxidant production. Electrophoretic mobility shift assay revealed that the ARE complex consisting of Nrf-1/2 and Maf proteins was activated 12~48 h. The rGSTA2 mRNA and protein levels were increased by SAAD. Activation of ARE and induction of rGSTA2 were both completely inhibited by PI3-kinase inhibitors. Inhibition of p38 MAP kinase by SB203580 prevented the ARE-mediated rGSTA2 induction. The results of this study showed that PI3-kinase might play an essential role in the ARE-mediated rGSTA2 induction by see or SAAD and that the dual MAP kinase pathways were responsible for the enzyme induction.

• 한국축산식품학회지
• /
• 제35권3호
• /
• pp.360-369
• /
• 2015

Milk proteins have many potential sequences within their primary structure, each with a specific biological activity. In this study, we compared and investigated the bioactivities of hydrolysates of the domestic (A, B) and imported (C, D) skim milk powders generated using papain digestion. MALDI-TOF analysis revealed that all milk powder proteins were intact, indicating no autolysis. Electrophoretic analysis of hydrolysates showed papain treatment caused degradation of milk proteins into peptides of various size. The antioxidant activity of the hydrolysates, determined using 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and total phenolic contents (TPC) assays, increased with incubation times. In all skim milk powders, the antioxidant activities of hydrolysates were highest following 24 h papain treatment (TPC: A, 196.48 μM GE/L; B, 194.52 μM GE/L; C, 194.76 μM GE/L; D, 163.75 μM GE/L; ABTS: A, 75%; B, 72%; C, 72%; D, 57%). The number of peptide derived from skim milk powders, as determined by LC-MS/MS, was 308 for A, 283 for B, 208 for C, and 135 for D. Hydrolysate A had the highest antioxidant activity and the most potential antioxidant peptides amongst the four skim milk powder hydrolysates. A total of 4 β-lactoglobulin, 4 α_s1-casein, and 56 β-casein peptide fragments were identified as potential antioxidant peptides in hydrolysate A by LC-MS/MS. These results suggest that domestic skim milk could have applications in various industries, i.e., in the development of functional foods.

• 대한의생명과학회지
• /
• 제13권4호
• /
• pp.287-291
• /
• 2007

The factors and mechanisms by infection of Oriental Tsutsugamushi caused disease are not well understood. The onset of tsutsugamushi disease is characterized by chilliness, fever, malaise, headache and generalized aching. Infection of tsutsugamushi is the cause of impairment of function of a major organ often complicate the picture and immediately change the prognosis for the worse. Tsutsugamushi disease is reported that this disease is characterized by the histopathogenesis of liver, kidney, heart, and lung, but the variation of biochemical components in serum of tsutsugamushi disease patient are not clear. We analyzed total protein (TP), albumin (AL), aspartic aminotranferase (AST), alanine aminotransferase (ALT), alkaline phosphotase (ALP), urea nitrogen (UN), creatinine (CRE), glucose (GLD), cholesterol (CHOL) and total bilirubin (TB) in sera of patients with tsutsugamushi disease. In comparison with reference, total protein and albumin were abnormally decreased in 19.6% and 39.2% of patients, respectively. AST, ALT, ALP, creatinine, UN, glucose, cholesterol and total bilirubin were abnormally increased in 94.1 %, 72.5%, 25.5%, 15.7%, 9.8%, 62.7%, 25.5% and 6.0% of patients, respectively. The patients showed abnormal relative rate of protein electrophoretic fractions to total protein in serum compared to them of reference were 43.1% (albumin), 12.9% ($\alpha_1$-globulin), 58.8% ($\alpha_2$-globulin), 60.8% ($\beta$-globulin) and 70.6% ($\gamma$-globulin), respectively. These data suggest that infection of Oriental Tsutsugamushi causes impairment of function of a major organ and abnormal serum protein electrophoresis fractions to tsutsugamushi patients.

• Restorative Dentistry and Endodontics
• /
• 제22권2호
• /
• pp.693-701
• /
• 1997

In 1992, Prevotella intermedia was shown to be comprised of another spoecies now known as Prevotella nigrescens. Strain ATCC 33563 is now designated the type strain of P. nigrescens while strain ATCC 25611 is remains the type strain of P. intermedia. The purpose of this study was to find the differences in protein profiles of P. intermedia and P. nigrescens, using sodium dodecyl sulfate polyacrylamide gel electrophoresis, which can be used for differentiation of those two species. A partial amino acid sequence of the 18.6 kDa protein band, which was specific in P. nigrescens, was also determined. The cellular proteins were extracted from the cell pellets of pure cultures of P. intermedia. and P. nigrescens by either sonication or being shaken continuously for 20 min at $21^{\circ}C$ with 1 % SDS or being boiled for 3 min with 1 % SDS. SDS-PAGE was performed according to the method of laemmli using either 12% (w/v) gels or 18% (w/v) gels. Results were as follows ; 1. The similar electrophoretic protein profiles were shown by 3 cellular protein extraction methods for each strain. (Fig. 1 and 2) 2. the 18.6 kDa band which was specific only in P. nigrescens could be used for the differentiation of P. intermedia. and P. nigrescens. (Fig. 1 and 2, Table 1) 3. A total of 4 different tryptic fragments from the 18.6 kDa protein were sequenced. the resulting amino acid sequences were fragment 1.GNPVNIGGEW, 2.FNVVR, 3.NYLT-VAPY, and 4.GGDNVTTYQVLPEIGYN. By comparison to the sequences of known proteins in the Swiss-Prot database and PIR database. 90 % matching between fragment 1 and serine hydroxymethyl transferase(P24060) in the Swiss-Prot, and 90% matching between fragment 1 and glycine hydroxymethyl transferase(S15203) in the PIR were shown, but the identity and function of the 18.6 kDa protein remains unknown.

• Applied Biological Chemistry
• /
• 제29권1호
• /
• pp.10-15
• /
• 1986

Cytokinin과 단백질의 결합을 간단하게 결정하는 방법으로서 전기 영동법을 시도하고, 대두의 잎단백질과 cytokinin의 결합여부, cytokinin에 대한 affinity가 있는 단백질의 종류와 상대적 affinity를 조사하였다. 검토된 전기 영동법은 cytokinin과 단백질의 결합뿐만 아니라, cytokinin에 대한 상대적 affinity를 동시에 검정할 수 있는 장점을 가지고 있었다. Ammonium sulfate 침전법, Sephadex G-25 chromatography, paper chromatography, 그리고 전기영동법으로 대두의 잎단백질 중에 BA와 결합하는 단백질이 있음을 확인할 수 있었다. 전기영동법으로 검정한 결과 BA와 결함하는 것이 3 group이 있고, 이중에서 전기영동 이동도로보아 분자량이 작은 단백질 분획과 전기영동 이동도 0.4부근의 분획은 BA에 대한 affinity가 비교적 낮은 반면, 이동도 $0.0{\sim}0.2$의 분획은 affinity가 큰 것으로 생각되었다.

• Journal of Microbiology and Biotechnology
• /
• 제24권3호
• /
• pp.386-393
• /
• 2014

Possible cross-transmission of hospital-associated enterococci between human patients, medical staff, and hospital environments has been extensively studied. However, limited information is available for veterinary hospital-associated Enterococcus isolates. This study investigated the possibility of cross-transmission of antibiotic-resistant enterococci between dog patients, their owners, veterinary staff, and hospital environments. Swab samples (n=465) were obtained from five veterinary hospitals in Seoul, Korea, during 2011. Forty-three Enterococcus strains were isolated, representing seven enterococcal species. E. faecalis and E. faecium were the most dominant species (16 isolates each, 37.2%). Although slight differences in the antibiotic resistance profiles were observed between the phenotypic and the genotypic data, our antibiogram analysis demonstrated high prevalence of the multiple drug-resistant (MDR) isolates of E. faecalis (10/16 isolates, 62.5%) and E. faecium (12/16 isolates, 75.0%). Pulsed-field gel electrophoretic comparison of the MDR isolates revealed three different clonal sets of E. faecalis and a single set of E. faecium, which were isolated from different sample groups or dog patients at the same or two separate veterinary hospitals. These results imply a strong possibility of cross-transmission of the antibiotic-resistant enterococcal species between animal patients, owners, veterinary staff, and hospital environments.

• 한국작물학회지
• /
• 제29권4호
• /
• pp.350-355
• /
• 1984

본 실험은 현재 줄기, 엽병 및 과실의 색에 의해 자경종, 황숙종 및 청경종으로 분류되어 있는 인삼의 변종들이 유전인자의 변이에 따른 것인지의 여부를 확인하기 위하여 국내 자경종, 황숙종의 종자와 2,3년생의 뿌리 그리고 청경종, 산양삼, 일본자경종, 황숙종, 미마끼와 미국삼의 2년생 뿌리를 사용하여 protein 과 esterase, phosphatase, GOT, LAP 및 peroxidase의 동위효소 표현형을 2∼30％ polyacrylamide gradient tube gel을 사용 전기영동법에 의해 관찰한 결과는 다음과 같았다. 1. 동일변종내에서 개체간 및 2년생 뿌리와 3년생 뿌리간에는 동일한 동위효소의 표현형을 가지고 있었으나 종자와 뿌리는 다른 표현형을 보였다. 2. 자경종과 황숙종의 종자는 protein 및 관찰한 모든 동위효소의 표현형간에 차이가 없었다. 3. 2년생 뿌리에서 국내인삼과 일본삼은 동일한 protein과 동위효소의 표현형을 보였으나 미국삼은 다른 표현형을 보였다. 따라서 본 실험결과로서는 국내인삼과 일본삼의 변종들은 모두 동일한 것으로 사료된다.

• 한국균학회지
• /
• 제26권2호통권85호
• /
• pp.163-172
• /
• 1998

국내외에서 수집된 Pleurotus속 13종 32균주와 종(species)이 밝혀지지 않은 4균주 등 모두 36균주의 동위효소 분석에 의한 종간 유연관계를 파악하기 위하여 본 실험을 수행하였다. Pleurotus속 균주들의 균사체에서 수용성 단백질을 추출하여 등전점 전기영동 isozyme polymorphic banding pattern법으로 여섯 종류의 동위효소 즉 esterase, glucosephosphate isomerase, leucine aminopeptidase, malate dehdrogenase, peroxidase, phosphoglucomutase를 분석하여 총 166개 밴드를 확인하였고 36개 느타리 균주의 종간 다형성을 관찰하였다. Esterase가 종간에 가장 다양한 밴드 pattern을 나타내어 종의 구별에 아주 유용한 동위효소라 생각되며, 다른 동위효소에서도 종간 다형성을 관찰할 수 있었다. 동위효소 분석 결과 유연관계가 가장 가까운 종은 P. florida와 P. sajor-caju로 유사도가 약 89%였으며, 연구에 이용된 총 13종간의 유사도는 약 77%였다. 분류상 논란이 많이 되는 P. ostreatus는 P. pulmonarius 종과 확실히 구별되었으며 P. florida도 독립된 종으로 판명되었다. 그러나 P. sapidus와 P. spodoleucus 균주들은 확실한 종의 판명이 어려웠다. Isozyme polymorphic banding pattern 결과와 현미경적인 형태 분류에 의한 종 판명 결과가 몇 종을 제의하고는 일치하였다. 동위효소를 이용한 분류는 매우 유용하다고 판단되었다.

• Applied Biological Chemistry
• /
• 제26권1호
• /
• pp.65-72
• /
• 1983

네가지 쌀단백군의 양적 관계는 주로 추출조건에 의하여 좌우되며 albumin 및 globulin의 추출양은 교반방법보다 추출온도에 의하여, glutelin은 사용용매의 종류와 pH에 의하여 추출양이 크게 좌우되었다. 적합한 쌀단백질의 추출로는 albumin 및 globulin은 0.5M-NaCl용액으로 $0{\sim}5^{\circ}C$에서, prolamin은 70%(v/v) ethanol용액으로 상온에서, glutelin은 0.1N acetic acid보다는 0.5% SDS-borate buffer (pH 8.3) 과 0.6% ${\beta}-mercaptoethanol$을 포함한 동일용액을 사용하는 것이 효과적이었다. 상기방법으로 추출한 각 단백군 구성성분의 분자량 분포는 albumin은 29,000, $18,000{\sim}15,000$ 및 12,000의 세가지이며 globulin은 27,000, 21,000 17,000 및 12,000의 4가지 이외에 고분자량의 단백질($10^5$이상)로 되어 있고 prolamin은 16,000, 0.5% SDS-borate buffer에 가용성인 glutelin은 35,000, 21,000, 17,000 및 15,000의 4가지와 이외에 globulin의 경우와 같이 고분자 단백질로 구성되어 있었다. Prolamin을 제외한 세 단백군은 각각 disulfide bond로 연결된 subunit를 가지고 있는 구성성분을 포함하고 있음을 알 수 있었다. SDS-glutelin은 Sephadex G-150 column chromatography에 의하여 세가지 fraction으로 분리할 수 있었다. Albumin과 globulin은 starch gel 전기영동상 산성 (pH 3.1)에서는 총 13개 및 21개의 band를 각각 나타내었고 염기성 (pH 8.95)에서는 총 4개 및 6개의 band를 확인할 수 있었다. 쌀품종들은 albumin과 globulin의 starch gel전기영동 pattern에 의하여 구별할 수 있었다. 산성조건에서는 특수 band의 존재 또는 band의 조합양상에 의하여 각 품종의 구별이 가능하였고 염기성조건에서는 각 품종별 구별은 할 수 없었으나 Japonica종을 Indica 또는 Japonica와 Indica교배종과 분리 식별할 수 있었다.