• Parasites, Hosts and Diseases
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• v.42 no.2
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• pp.57-60
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• 2004

A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

• Parasites, Hosts and Diseases
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• v.51 no.2
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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.

• Journal of Life Science
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• v.8 no.5
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• pp.598-603
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• 1998

A marine microbe, Vibrio anguillarum was isolated from fish and studied for its concerning pathogenic substance of hemolysin. Purification of hemolysin was achieved by the procedure of ammonium sulfate precipitation from cul-ture filtrate, DEAE-cellulose chromatography, and G-200 gel filtration with 36 fold of purification and 2.3% yield. The molecular weight of the purified hemolysin was 38,000 dalton by SDS-PAGE. The purified hemolysin was stable at pH 6-9, below 45$^{\circ}C$, and up to 1% of NaCl, respectively. $Ca^{2+}, Cu^{2+}, Zn^{2+}, Fe^{2+}$ inhibited the hemolytic activity whereas EDTA and $Mg^{2+}$ did not.

• Journal of Ginseng Research
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• v.7 no.2
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• pp.156-162
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• 1983

Stability of nitrogen compounds in the water-extract of ginseng has been investigated in terms of concentration and SDS-PAGE protein pattern as functions of times and temperature. The concentration of free amino-nitrogen in the extract of fresh ginseng varied with time at various temperatures. Especially at temperature over 90$^{\circ}C$, it showed a tendency of steady declease at a considerably fast rate. In the case of red ginseng which had been commercially processed and stocked for 1-7 years, it was found that the free amino-N concentration diminished gradually depending on the length of stock period of samples. This result may be comparative to another observation that the amount and density distribution of precipitate from extract varied with the lapse of stock time, based on accelerated sedimentation test. The number of SDS-PAGE protein bands was observed to be at least seven when determined with the extract of fresh ginseng. However, it tended to reduce finally to be one band when the extract was kept at a rather high temperature for a long time.

• Journal of Food Hygiene and Safety
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• v.14 no.3
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• pp.233-237
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• 1999

The bacteriocin produced by Lactobacillus sp. GM7311 was purified by sequential steps including n-propanol/acetone treatment, CM-cellulose chromatography, and gel filtration on Sephacryl HR-100. The relative activity of bacteriocin increased 493-fold after final purification step with a recovery of 8.3%. Two protein bands of ca. 8,200 and 2,500 were detected by SDSPAGE of bacteriocin purified through CM-cellulose and sephacryl HR-100 chromatography and both of them had bacteriocin activity.

• Korean journal of food and cookery science
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• v.20 no.3
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• pp.256-264
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• 2004

The effects of irradiation on the functional and structural characteristics of soy protein isolates were studied. Soymilk was irradiated at 1, 5, and l0kGy, after which soy protein isolates were prepared. The functional properties of soy protein isolates were examined including solubility, emulsion capacity and stability, foam capacity and stability, structural properties as represented by SDS-PAGE pattern, and secondary and tertiary structures. The solubility and emulsion capacity were increased by radiation treatment at 1kGy however the values were adversely affected again as dosage was increased above 5kGy. As irradiation dosage increased, an increase of foaming capacity at 1kGy and a decreasing turnover afterwards were also noted in foaming capacity, although the differences were not statistically significant. The SDS-PAGE pattern showed fragmentation and aggregation of protein molecules as affected by irradiation in proportion to the dosage increase. The results of CD and fluorescence spectroscopy revealed increased aperiodic structure contents with the dosage increase. It was assumed that irradiation dosagefrom 5 to l0kGy could initiate minimal denaturation of protein in various foods compared to general heat treatment.

• KSBB Journal
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• v.14 no.3
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• pp.322-327
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• 1999

The cholesterol oxidase(EC.1.1.3.6) produced from Streptomyces sp. No.4 which isolated from soil was purified and investigated for the enzymatic properties. The enzyme was purified specifically by cholesterol affinity column chromatography with a yield of 28.3%. The purified enzyme showed a single polypeptide on SDS-PAGE and the molecular weight was estimated to be 60,000 daltons. The enzyme activity was strongly inhibited by metal ions such as $HgCl_2$ and $CuSO_4$. Dithiothreitol and mercaptoethanol inhibited the enzyme activity at concentration of 1mM. The Michaelis constant(Km) for cholesterol was found to be 1.38mM by Lineweaver-Burk plot analysis. Amino acid analysis showed that the enzyme protein was composed of 416 amino acid residues including 52moles of glycine and 19moles of tryptophane.

• Culinary science and hospitality research
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• v.11 no.2
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• pp.110-124
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• 2005

This study was conducted to investigate the proteolytic properties of Sarcodon aspratus on meat proteins. The analytical condition for the measurement of enzyme activity was determined and the effect of Sarcodon aspratus on beef protein and its fractions were determined by SDS-PAGE and the spectrophotometric method respectively. Optimum temperature and pH of Sarcodon aspratus were $73~78^{\circ}C$ and pH 8 respectively. However, enzyme tended to be denatured at $50^{\circ}C$ for 10 min incubation. Proteolytic activity of Sarcodon aspratus was higher than of kiwi and pear 66 and 990 times by dry weight respectively. It appeared that proteolytic activity of Sarcodon aspratus toward beef protein by SDS-PAGE was prominent when compared to that of kiwi and bromelain. Furthermore, Sarcodon aspratus showed the highest proteolytic activity toward all the beef protein fractions, which was followed by collagenase and bovine protease. Transmission electron microscopy showed the muscle fiber started to be degraded when treated with Sarcodon aspratus(1,000 unit) for 10min at $25^{\circ}C$. No distinct sarcomere, A-band, or z-line was observed when treated with Sarcodon aspratus for 60min at the same condition.

• BMB Reports
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• v.34 no.4
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• pp.310-315
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• 2001

The uneven distribution of acidic and basic chitinases in different parts of rice seed, and also the characterization of hull-specific chitinases, are reported here. After extraction of chitinases from polished rice, bran, and rice hulls, the chitinases were separated into acidic and basic fractions, according to their behavior on an anion exchanger column. Both fractions from different parts of rice seed showed characteristic activity bands on SDS-PAGE that contained 0.01% glycol chitin. The basic chitinases from rice hulls were further purified using chitin affinity chromatography. The chitinase, specific to rice hulls (RHBC), was 88-fold purified with a 1.3% yield. RHBC has an apparent molecular weight of 22.2 kDa on SDS-PAGE. The optimal pH and temperature were 4.0 and $35^{\circ}C$, respectively. With [$^3H$]chitin as a substrate, RHBC has $V_{max}$ of 13.51 mg/mg protein/hr and $K_m$ of 1.36 mg/ml. This enzyme was an endochitinase devoid of ${\beta}$-1,3-glucanase, lysozyme, and chitosanase activities.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.5
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• pp.721-724
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• 2002

Postmortem Proteolysis of breast (BM) and leg (LM) muscles in Chiayi native chickens at $5^{\circ}C$ were compared. Myofibrils were purified from BM and LM samples that were randomly taken from carcasses after 0, 1, 3, 7 and 14 days of storage at $5^{\circ}C$. Fragmentation of myofibrils were determined, and degradation of myofibrillar proteins were analyzed by the SDS-PAGE and western blots. The results showed that myofibril fragmentation index (MFI) was significantly (p<0.05) higher in BM than in LM samples. Disappearance of titin and nebulin and appearance of the 30 kDa component were more rapidly as seen on SDS-PAGE in BM than in LM samples. Western blots labeled with a monoclonal antibody to desmin also demonstrated that desmin degraded more quickly in BM samples. Our data suggested that postmortem proteolysis occurred more rapidly in breast muscles in Chiayi native chickens.