• 한국식품영양과학회지
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• 제24권4호
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• pp.587-593
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• 1995

택사로부터 황산암모늄 분별 침전, DEAE-cellulose ion exchange chromatography, Sephadex G-150 chromatography 등의 방법을 이용하여 렉틴을 분리, 정제하였다. 정제한 렉틴의 분자량은 gel filtration법에 의해 측정한 결과 90,500 dalton이었으며, SDS polyacrylamide gel 전기영동을 실시한 결과 subunit 분자량은 각각 42,000, 27,000, 22,500 dalton으로 나타나므로 택사 렉틴이 heterotrimer임을 알았다. 정제된 택사 렉틴은 사람 적혈구의 경우 모든 혈액형에 대하여 응집현상을 나타내었으며 돼지, 쥐 및 개 등의 적혈구에 대해서도 모두 응집현상이 나타나 혈구 비특이성임을 보여주었다. 또한 이 렉틴은 sialic acid, glucose, ribose, sucrose, lactose, galactose 등 당에 의해서, 그리고 $Hg^{2+},\;Fe^{2+},\;Cu^{2+}$ 이온 등에 의해 적혈구 응집력이 저해되었다.

• 한국식품과학회지
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• 제12권1호
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• pp.24-33
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• 1980

한국산 두류 중 대두를 비롯한 팥, 녹두, 강낭콩에 대하여 trypsin 저해(沮害) 활성(活性)과 적혈구(赤血球) 응집(凝集) 활성(活性)을 측정하고 이들을 가열 처리에 의한 파괴 효과를 실험한 결과는 다음과 같다. 1. 각 두류의 건물(乾物) 중량 당 저해(沮害) 활성도(活性度) (% [TU] inhibited/mg)는 대두가 가장 높아 79.9를 나타낸 반면 팥, 강낭콩은 각각 46.4, 43.2로 대두의 반정도이었으며 녹두는 17.7로 제일 적었다. 이를 단백질 중량당으로 환산하면 팥이 제일 높아 222를 보였고 그 다음이 대두 194, 강낭콩 170의 순이었으며 녹두는 제일 낮아 75를 보였다. 2. 열에 의한 trypsin 저해(沮害) 활성도(活性度)의 파괴율은 팥, 녹두가 각각 $85{\sim}87,\;87{\sim}94\;%$로 높았고 강낭콩, 대두는 각각 $76{\sim}79,\;67{\sim}72\;%$로 낮았다 그러나 boiling과 autoclaving 간의 가열 방법에 따른 차이는 발견할 수 없었다. 3. 각 두류의 토끼 혈액에 대한 적혈구(赤血球) 응집(凝集) 활성(活性)(hemagglutinating unit/g)은 팥에서는 나타나지 않았고 강남콩이 제일 많아 건물 1g당 48,300을 보인데 비해 대두는 18,000, 녹두는 136이었다. 이를 단백질 중량당으로 환산하면 마찬가지 경향을 보여 강남콩 190,600, 대두 43,700, 녹두 581로 나타났다. 이들 두류의 적혈구(赤血球) 응집(凝集) 활성(活性)은 가열 처리에 의하여 모두 파괴되었다.

• Food Science and Biotechnology
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• 제18권5호
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• pp.1173-1179
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• 2009

Trypsin inhibitors and lectins are defense proteins produced by many organisms. From Chinese 'Large Black Soybeans', a 60 kDa lectin and a 20 Da trypsin inhibitor (TI) were isolated using chromatography on Q-Sepharose, Mono Q, and Superdex 75. The TI inhibited trypsin and chymotrypsin with an $IC_{50}$ of 5.7 and $5{\mu}M$, respectively. Trypsin inhibitory activity of the TI was stable from pH 3 to 13 and from 0 to $65^{\circ}C$. Hemagglutinating activity of the lectin was stable from pH 2 to 13 and from 0 to $65^{\circ}C$. The TI was inhibited by dithiothreitol, signifying the importance of disulfide bond. The TI and the lectin inhibited HIV-1 reverse transcriptase ($IC_{50}$=44 and $26{\mu}M$), and proliferation of breast cancer cells ($IC_{50}$=42 and $13.5{\mu}M$) and hepatoma cells ($IC_{50}$=96 and $175{\mu}M$). The hemagglutinating activity of the lectin was inhibited most potently by L-arabinose. Neither the lectin nor the TI displayed antifungal activity.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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• pp.161-162
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• 2000

Metalloprotease produced in Vibrio mimicus, in which zinc is an essential meta ion for catalytic activity, degrades a variety of biologically important substances including human collagen, several complement components, and immunoglobulin. For gene overexpression and convenient purification, VMC gene was constructed in pET22b(+) expression vector by using of PCR. (omitted)

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.612-619
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• 2000

To investigate the biochemical properties of V. mimicus metalloprotease, whose gene was isolated previously from Vibrio mimicus ATCC33653, overexpression and purification were attempted. The 1.9 kb of open reading frame was amplified by PCR from pVMC193 plasmid which ligated the VMC gene with pUC19 and introduced into Escherichia coli BL21 (DE3) using the overexpression vector, pET22b (+). The overexpressed metalloprotease (VMC) was purified with Ni-NTA column chromatography and characterized with various protease inhibitors, pHs, temperatures, and substrates. The purified VMC showed the proteolytic activity against gelatin, soluble and insoluble collagens, and synthetic peptides. Unlike the observations made with all metalloproteases originated from other Vibrio sp., the VMC did not hydrolyze the casein. The proteolytic activity was critically decreased when the VMC was treated with metal chelating reagents, such as EDTA, 2,2-bipyridine, and 1, 10-phenanthroline. In particular, the 71 kDa VMC exhibited the hemagglutinating activity against human erythrocyte. As the purified VMC was treated with $CuCl_2$ and $NiCl_2$ for the chemical modification of metal binding, the proteolytic activity and hemagglutinating activity were profoundly influenced. The multialignment analysis made on the reported Vibrio metalloproteases showed the difference of amino acid sequence similarity between the two distinctive classes of Vibrio metalloproteases.

• 한국동물위생학회지
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• 제17권3호
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• pp.174-180
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• 1994

The disease syndrome characterized by the acute vomiting and diarrhea with high mortality had been greatly epidemic in Kyungbuk West Area since March 1990 and it was followed serologically for the classification of the agent. The agent present in feces of dogs associated with this syndrome had characteristic feature in agglutinating pig red blood cells that was specifically inhibited by anti-CPV reference dog serum. This also showed the serological identity with the reference CPV antigen in Hemagglutinating inhibition test. The result obtained were summarized as follows : 1. During 5 years(March. 1990∼September. 1994), 1,470 dogs were investigated on the actual condition of CPV infections. The Infection rate of CPV from dogs was 62.5% and mortality rate was 59.8%. 2. Among 24 fecal samples collected from the dogs with enteric disease, all showed the hemagglutinating activity to porcine erythrocyte ranging from 40 to 5,120 of HA titers. 3. Among 12 sera samples collected from the dogs with enteric disease, all showed the serological identity with the reference CPV antigen from 5 to 5,120 of HI titers. 4. Bacteriologic examination of fecal specimens resulted in the isolation of pathogeric bacteria such as Staphylococcus sp, Streptococcus sp, Escherichia coli and Bacillus. Cultures for salmonella sp and Clostridium remaind negative. 5. The prevalence and identification of internal parasites were determined by fecal examination using the floatation methods. From 20 fecal samples 12(60.0%) were isolated and their species were Toxacara canis, Toxascaris leonina, and coccidium.

• Archives of Pharmacal Research
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• 제20권4호
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• pp.306-312
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• 1997

We attempted to isolate and characterize the lectins from stem and leaves of Korean mistletoe (Viscum album var. coloratum) by affinity chromatography. Lectin I was isolated only from stem. Lectin II was not isolated from Korean mistletoe, whereas lectin III was isolated from the stem and leaves. The hemagglutinating activity of lectin I was 16HU and inhibited by D-galactose, lactose, and N-acetyl-D-galactosamine. The lectin I has molecular weight of 60, 000D being composed of two basic subunits with molecular weights of 32, 000D and 28, 000D which are linked by a disufide bond. The lectin III from stem has molecular weight of 66, 000D being two basic subunits which have molecular weights of 34, 000D and 29, 000D and are linked by a disufide bond. The activity of lectin I was stable at the pH range of 4.00-8.50 and at a wide range of temperature (0-42.deg. C). The lectin I showed more potent mitogenic activity to murine lymphocytes than concanavalin A.

• BMB Reports
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• 제36권2호
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• pp.214-222
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• 2003

A lectin named AAL has been purified from the fruiting bodies of the edible mushroom Agrocybe aegerita. AAL consisted of two identical subunits of 15.8 kDa, its pI was about 3.8 determined by isoelectric focusing, and no carbohydrate was discerned. Being treated by pyrogultamate aminopeptidase, the blocked N-terminus of AAL was sequenced as QGVNIYNI. AAL agglutinated human and animal erythrocytes regardless of blood type or animal species. Its hemagglutinating activity was unaffected by acid or alkali treatment and demetalization or addition of divalent metals $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$. AAL was toxic to mice: its LD50 was 15.85 mg per kilogram body weight by intraperitoneal injection. In this study, two novel activities of AAL were proved. It showed inhibition activity to infection of tobacco mosaic virus on Nicotiana glutinosa. The result of IEF suggested that AAL attached to TMV particles. Mycelia differentiation promotion was the other interesting activity. AAL promoted the differentiation of fruit body primordia from the mycelia of Agrocybe aegerita and Auricularia polytricha. AAL antiserum was prepared and immunologically cross-reactived with several proteins from five other kinds of mushrooms. These results suggested that AAL probably was a representative of a large protein family, which plays important physiological roles in mushroom.

• Archives of Pharmacal Research
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• 제14권3호
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• pp.207-216
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• 1991

Pseudixus japonicus agglutinin (PJA) was isolated. And its characteristics were compared with those of concanavalin A (Con A). PJA is a glycopritein composed of 49.3% carbohydrate and 50.7% protein which had relatively high percentages of glutamic acid, aspartic acid and phenylalanine residues. The hemagglutinating activity of PJA was approximately one-eighth of that of Con A when tested with mouse crythrocytes. PJA failed to simulate the proliferation or transformation of human and mouse lymphocytes in contratst to Con A. PJA and Con A showed cytotoxicities against SNU-1 (human stomach cancer cells), SNU-CI (human colon cancer cells) and mouse Sarcoma 180 cells when tested by 3-(4, 5-dimethyl thiazol-2-yl)2. 5-diphenyl tetrazolium bromide (MIT) colorimetric assay. The antitumor activity of the lectin in vivo was also tested in Sarcoma 180 bearing mice. There was no significant difference in prologation of lifc span of the mice after the treatment with PJA and Con A for 10 consecutive days.

• BMB Reports
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• 제40권3호
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• pp.358-367
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• 2007

A novel mannose-binding tuber lectin with in vitro antiproliferative activity towards human cancer cell lines and antiviral activity against HSV-II was isolated from fresh tubers of a traditional Chinese medicinal herb, Typhonium divaricatum (L.) Decne by a combined procedure involving extraction, ammonium sulfate precipitation, ion exchange chromatography on DEAE-SEPHAROSE, CM-SEPHAROSE and gel-filtration on sephacryl S-200. The apparent molecular mass of the purified Typhonium divaricatum lectin (TDL) was 48 kDa. TDL exhibits hemagglutinating activity toward rabbit erythrocytes at 0.95 $\mu$g/ml, and its activity could be strongly inhibited by mannan, ovomucoid, asialofetuin and thyroglobulin. TDL showed antiproliferative activity towards some well established human cancer cell lines, e.g. Pro-01 (56.7 $\pm$ 6.8), Bre-04 (41.5 $\pm$ 4.8), and Lu-04 (11.4 $\pm$ 0.3). The anti-HSV-II activity of TDL was elucidated by testing its HSV-II infection inhibitory activity in Vero cells with $TC_50$ and $EC_50$ of 5.176 mg/ml and 3.054 $\mu$g/ml respectively. The full-length cDNA sequence of TDL was 1145 bp and contained an 813-bp open reading frame (ORF) encoding a 271 amino acid precursor of 29-kDa. Homology analysis showed that TDL had high homology with many other mannose-binding lectins. Secondary and three-dimensional structures analyses showed that TDL is heterotetramer and similar with lectins from mannose-binding lectin superfamily, especially those from family Araceae.