• 제목/요약/키워드: Protein fractions

검색결과 783건 처리시간 0.03초

18α-Glycyrrhetinic acid의 위암 세포 사멸 효과에 관한 연구 (18α-Glycyrrhetinic acid induces apoptosis of AGS human gastric cancer cells)

  • 김정남;김병주
    • 대한한의학방제학회지
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    • 제28권1호
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    • pp.63-70
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    • 2020
  • Objectives : The purpose of this study was to investigate the anti-cancer effects of 18α-Glycyrrhetinic acid (18α-GA), a hydrolyzed metabolite of glycyrrhizin, in AGS human gastric adenocarcinoma cells. Methods : We used human gastric adenocarcinoma cell line, AGS cells. We examined cell death by MTT assay and caspase 3 and 9 assay with 18α-GA. To examine the inhibitory effects of 18α-GA, sub-G1 analysis was done the AGS cells after 24 hours with 18α-GA. Also, to investigate the inhibitory mechanisms of 18α-GA, mitogen-activated protein kinase pathways and reactive oxygen species (ROS) generation were examined. Results : 1. 18α-GA inhibited the growth of AGS cells in a dose-dependent fashion. 2. Sub-G1 fractions were significantly and dose-dependently increased by 18α-GA. 3. 18α-GA increased the caspase 3 and 9 activities in AGS cells. 4. 18α-GA inhibited proliferation of AGS cells via the modulation of c‑Jun N‑terminal kinase (JNK) signaling pathways, which results in the induction of apoptosis. 5. 18α-GA enhanced ROS accumulation in AGS cells. Conclusions : Our findings provide insight into unraveling the effects of 18α-GA in human gastric adenocarcinoma cells and developing therapeutic agents against gastric cancer.

Overexpression, Purification, and Characterization of $\beta$-Subunit of Group II Chaperonin from Hyperthermophilic Aeropyrum pernix K1

  • Shin, Eun-Jung;Lee, Jin-Woo;Kim, Jeong-Hwan;Jeon, Sung-Jong;Kim, Yeon-Hee;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • 제20권3호
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    • pp.542-549
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    • 2010
  • In the present study, overexpression, purification, and characterization of Aeropyrum pernix K1 chaperonin B in E. coli were investigated. The chaperonin $\beta$-subunit gene (ApCpnB, 1,665 bp ORF) from the hyperthermophilic archaeon A. pernix K1 was amplified by PCR and subcloned into vector pET21a. The constructed pET21a-ApCpnB (6.9 kb) was transformed into E. coli BL21 Codonplus (DE3). The transformant cell successfully expressed ApCpnB, and the expression of ApCpnB (61.2 kDa) was identified through analysis of the fractions by SDS-PAGE (14% gel). The recombinant ApCpnB was purified to higher than 94% by using heat-shock treatment at $90^{\circ}C$ for 20 min and fast protein liquid chromatography on a HiTrap Q column step. The purified ApCpnB showed ATPase activity and its activity was dependent on temperature. In the presence of ATP, ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}$ and $50^{\circ}$, respectively. Specifically, the activity of malate dehydrogenase (MDH) at $85^{\circ}$ was greatly stabilized by the addition of ApCpnB and ATP. Coexpression of pro-carboxypeptidase B (pro-CPB) and ApCpnB in E. coli BL21 Codonplus (DE3) had a marked effect on the yield of pro-CPB as a soluble and active form, speculating that ApCpnB facilitates the correct folding of pro-CPB. These results suggest that ApCpnB has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.

Aflatoxin $B_1$의 검출을 위한 효소면역측정법의 개발 (Development of an Enzyme-Linked Immunosorbent Assay for the Iletection of Aflatoxin $B_1$)

  • 손동화;박애란;서병철;김진철;이인원;남영중;허우덕
    • 한국미생물·생명공학회지
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    • 제20권2호
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    • pp.225-232
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    • 1992
  • 효소면역측정법에 의한 aflatoxin $B_1(AFB_1)$의 정량법을 개발하기 위하여, 항체를 생산.정제하여 분석법을 확립하고 직접법과 간접법(direct/indirect competitive ELISA)의 특성 및 문제점을 비교. 검토하였다. Bovine serum albumin(BSA)을 carrier protein으로 한 $AFB_1$-1-(O-carboxymethyl)oxime -BSA를 토끼에 면역하여 항$AFB_1$항혈청을 생산하였다. 정량 침강반응에 의하여 항혈청으로부터 항BSA항체를 제거하고 황산암모늄 침전법 및 EDAE-Sephadex A-50 이온교환 크로마토그래피를 통하여 순도 높은 IgG항체를 정제하여 항$AFB_1$항체를 사용하였다.

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(E)-3-(3-methoxyphenyl)-1-(2-pyrrolyl)-2-propenone displays suppression of inflammatory responses via inhibition of Src, Syk, and NF-κB

  • Kim, Yong;Jeong, Eun Jeong;Han Lee, In-Sook;Kim, Mi-Yeon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • 제20권1호
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    • pp.91-99
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    • 2016
  • (E)-3-(3-methoxyphenyl)-1-(2-pyrrolyl)-2-propenone (MPP) is an aldol condensation product resulting from pyrrole-2-carbaldehyde and m- and p- substituted acetophenones. However, its biological activity has not yet been evaluated. Since it has been reported that some propenone-type compounds display anti-inflammatory activity, we investigated whether MPP could negatively modulate inflammatory responses. To do this, we employed lipopolysaccharide (LPS)-stimulated macrophage-like RAW264.7 cells and examined the inhibitory levels of nitric oxide (NO) production and transcriptional activation, as well as the target proteins involved in the inflammatory signaling cascade. Interestingly, MPP was found to reduce the production of NO in LPS-treated RAW264.7 cells, without causing cytotoxicity. Moreover, this compound suppressed the mRNA levels of inflammatory genes, such as inducible NO synthase (iNOS) and tumor necrosis factor (TNF)-${\alpha}$. Using luciferase reporter gene assays performed in HEK293 cells and immunoblotting analysis with nuclear protein fractions, we determined that MPP reduced the transcriptional activation of nuclear factor (NF)-${\kappa}B$. Furthermore, the activation of a series of upstream signals for NF-${\kappa}B$ activation, composed of Src, Syk, Akt, and $I{\kappa}B{\alpha}$, were also blocked by this compound. It was confirmed that MPP was able to suppress autophosphorylation of overexpressed Src and Syk in HEK293 cells. Therefore, these results suggest that MPP can function as an anti-inflammatory drug with NF-${\kappa}B$ inhibitory properties via the suppression of Src and Syk.

치즈 및 된장에서의 쓴 맛 펩타이드 특성 (Characteristics of Bitter Peptides from a Cheese and a Soybean Paste)

  • 김수호;이형주
    • 한국식품과학회지
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    • 제17권4호
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    • pp.276-282
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    • 1985
  • 단백발효식품에서의 쓴맛 펩타이드의 특성을 규명하기 위하여 여러 시료로부터 소수성 펩타이드를 2 1(v/v) chloroform-methanol로 추출하였다 이 중 모짜렐라치즈와 된장으로부터 얻은 쓴맛 텝타이드 추출물을 다시 Sephadex C-25 겔 크로마토그라피한 결과 분획 I II, III을 얻었는데 이들 시료 자체와 각각의 크로마토그라피 분획을 구성하고 있는 펩타이드의 아미노산 조성을 분석하여 평균소수도를 계산하였다. 시료들 자체는 쓴 맛을 나타내지 않았으나 용매로 추출된 소수성 펩타이드 분획은 강한 쓴 맛을 나타내었다. 시료의 용매추출 수율은 총 질소 함량의 0.08내지 62.50%의 범위로 나타났다. 모짜렐라치즈 질소물의 평균 소수도는 1.376cal/mole, 용매추출 분획은 1.623cal/mole. 젤 크로마토그라피 분획 I은 1,797cal/mole, 분적 II 는 2,454cal/mole, 분획 III은 1,559cal/mole 이었다. 된장의 경우 된장 자체, 용매추출 분획, 겔 크로마토그라피 분획 I, II, III의 평균소수도는 각각 1,229. 1,654, 1900, 2016, 998cal/mole 이었다. 쓴맛 펩타이드에서 중요한 아미노산을 leucine, Phenylal anine. p-roline, valine으로 나타났다.

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오미자(五味子)의 일반성분(一般成分), 유기산(有機酸) 및 Anthocyanin 색소(色素)에 관하여 (On the Proximate composition, Organic acids and Anthocyanins of Omija, Schizandra chinensis Baillon)

  • 김경임;남주형;권태완
    • 한국식품과학회지
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    • 제5권3호
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    • pp.178-182
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    • 1973
  • 오미자(五味子) 열매의 일반성분(一般成分)을 살폈고, 유기산(有機酸)을 정량(定量)하였으며, 색소(色素)를 분획(分劃)하여 일부(一部) 동정(同定)하였다. 1. 숙성(熟成)한 오미자(五味子)에는 수분(水分) 84.2%, 지방(脂肪) 0.9%, 환원당(還元糖) 10.9%, 그리고 단백질(蛋白質)이 1.1% 함유(含有)되었으며, Total anthocyanin의 함량(含量)은 168mg%이다. 2. 오미자(五味子) 중(中) 확인(確認)된 유기산(有機酸)의 함량(含量)은 5.1%로서 citric acid 3.5%, malic acid 1.4%, 그리고 succinic acid 0.2%로 조성(組成)되었다. 3. 오미자(五味子)의 색소(色素)를 polyvinylpyrrolidone column에 통과(通過)시켜서 3개(個)의 획분(劃分)을 분리(分離)하였고, 그 중 80%를 차지하는 anthocyanin을 peonodin 3-glucoside로 잠정적으로 동정(同定)하였다.

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폐경 전·후 여성들의 체위, 영양소 섭취 상태 및 혈중 지질과의 관련성에 관한 연구 (Correlation of Anthropometric Data, Nutrient Intakes and Serum Lipids in Premenopausal and Postmenopausal Women)

  • 최윤희;송태희
    • 한국식품영양학회지
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    • 제26권3호
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    • pp.476-484
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    • 2013
  • This study was conducted to investigate the anthropometric data, nutrient intakes and serum profiles in premenopausal and postmenopausal women living in Gyeonggi-do Province, Republic of Korea. The subjects were 49 premenopausal women and 63 postmenopausal women who are not taking any hormone or cardiovascular drugs. Anthropometric measurements were taken by a trained practitioner and the data for dietary intakes were obtained by a 24-hour recall method. Serum samples were collected and analyzed for the total cholesterol, triglyceride (TG) and lipoprotein fractions. The mean age of the premenopausal women was $45.17{\pm}3.28$ years and that of the postmenopausal women was 2$62.5{\pm}4.14$ years. The height and weight were $157.86{\pm}$4.35 cm, $58.75{\pm}6.01$ kg in premenopausal women and $156.42{\pm}3.62$ cm, $57.63{\pm}5.38$ kg in postmenopausal women, respectively. WHR (waist hip ratio) in postmenopausal women was significantly higher than that of premenopausal women (p<0.05). There were no differences between the pre-and postmenopausal women in the intakes of energy, protein, fat, Ca, Fe, vitamin A, vitamin $B_1$, vitamin $B_2$, niacin, vitamin C and vitamin E. However carbohydrate and Na intakes in postmenopausal women were significantly higher than those of premenopausal women. In postmenopausal women, Ca intake was below and Na intake was extremely high considering KDRIs (Dietary Reference Intakes for Koreans). Serum triglyceride in postmenopausal women was positively correlated with age, BMI (body mass index) and WHR. Serum total cholesterol and triglyceride in postmenopausal women showed significantly negative correlations with fiber intake. These results suggest that it is necessary to help postmenopausal women maintain a healthy body weight. Postmenopausal women needs to increase Ca (calcium) intakes and diet quality by decreasing the intakes of Na (sodium). In addition, an adequate intakes of fiber is recommended for postmenopausal women to prevent cardiovascular disease.

Production of Iron-Binding Peptides from Colostral Whey by Enzymatic Hydrolysis

  • Kim, Sang-Bum;Ku, Min-Jung;Cho, Won-Mo;Ki, Kwang-Seok;Kim, Hyeon-Shup;Nam, Myoung-Soo
    • 한국축산식품학회지
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    • 제30권6호
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    • pp.923-929
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    • 2010
  • Colostral whey prepared from colostrum (pooled from first six post-partum milkings) was heated for 10 min at $100^{\circ}C$ Heated colostral whey was incubated with 1% enzymes (protein equivalent basis) for 15, 30, 60, 90, and 120 min at $50^{\circ}C$. Papain, pepsin, trypsin, and alcalase produced different degrees of hydrolysis (DH), 10.66%, 12.42%, 10.83%, and 25.31%, respectively, at an incubation time of 120 min. The SDS-PAGE reveals that significant amounts of bovine serum albumin (BSA), ${\beta}$-lactoglobulin (${\beta}$-LG), and ${\alpha}$-lactalbumin (${\alpha}$-LA) survived papain digestion. In contrast, pepsin completely removed BSA but not ${\beta}$-LG present in heated colostral whey. Alcalase completely eliminated BSA, ${\beta}$-LG, and ${\alpha}$-LA. This differential hydrolysis was confirmed by reversed-phase HPLC analysis. Using ion-exchange chromatography, fraction-1 (F-1) was obtained from alcalase hydrolysate at a NaCl gradient concentration of 0.25 M. Reversed-phase HPLC chromatograms of alcalase F-1 showed numerous small peaks, which probably indicate that a variety of new peptides were produced. Iron content of alcalase F-1 was 28.94 ppm, which was the highest among all enzyme fractions, whereas iron content of colostral whey was 36.56 ppm. Main amino acids contained in alcalase F-1 were Thr (15.45%), Glu (14.12%), and Ser (10.39%). Therefore, alcalase can be used to generate good iron-binding peptides in heated colostral whey, and the resulting iron-binding peptides could be suitable as a value-added food ingredient for food supplements.

미나리과 산채의 아질산염 소거능 (Nitrite Scavenging Effect of Umbelliferaeceae)

  • 노광석;양미옥;조은자
    • 한국식품조리과학회지
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    • 제18권1호
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    • pp.8-12
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    • 2002
  • 미나리과 산채인 참나물, 신선초, 방풍의 조단백은 22.99~31.00%, 조지방은 3.51~5.08%였으며, $\beta$-carotene은 5.40~19.81 mg%, ascorbic acid는 미량 함유하였고, Selenium은 모든 시료에서 검출되지 않았다. 총 페놀 함량은 617.87~875.77 mg%를 나타내었으며, 이 중 방풍이 가장 높은 함량을 보였다. 참나물, 신선초, 방풍의 지방산 조성은 전 지방산 중 linoleic acid와 linolenic acid가 66.46~77.44%를 차지하였다. 아질산염 소거능은 참나물, 신선초, 방풍 모두에서 메탄을 가용성 획분이 수용성 획분에 비하여 높은 소거능을 보였고, pH 1.2〉 4.2〉6.0 순으로 소거능이 높았다. 방풍의 메탄을 가용성 획분의 아질산염 소거능이 pH 1.2에서 95.3%로 가장 높았으며, 다음이 참나물, 신선초 순이었다. 총 페놀과 아질산염 소거능은 높은 상관관계를 나타내었다.

Camembert Cheese 숙성 중 Casein의 변화에 관한 연구 (Changes in Caseins during the Ripening of Camembert Cheese)

  • 이지환;이수원;정재홍;양융
    • 한국식품과학회지
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    • 제22권3호
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    • pp.337-342
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    • 1990
  • P. caseicolum V.B.와 복합유산균을 사용하여 제조한 Camembert cheese를 45일간 숙성시키면서 카제인 변화와 그에 따른 기호성을 조사하였다. 숙성과정 중 수용성 질소, 비카제인태 질소, 비단백태 질소량은 증가하였다. pH 4.6-insoluble casein의 전기영동 결과 숙성 10일에는 4개의 band에서 45일에 12개의 band로 분리되었는데 ${\alpha}_{s1}-casein$은 숙성 17일에 완전히 분해되었으며, ${\beta}-casein$도 숙성 45일 동안 거의 대부분이 분해되었다. 또한 pH 4.6-soluble casein fragments의 gel filtration에서 숙성 10일에는 3개의 fraction, 24일까지는 4개의 fraction이 나타났고 31일부터는 5개의 fraction으로 분별되었다. 관능검사 결과 숙성 31일된 Camembert cheese의 기호성이 제일 높았으며, 숙성이 진행됨에 따라 hardness는 감소하였고 관능적 기호성과는 0.1%의 유의수준으로 높은 상관관계를 나타냈다.

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