1.THE ANTI-TUMOR EFFECT OF THE PROTEOGLYCAN FROM ABALONE VISCERA AGAINST H_(22) CELL LINE IN VIVO
Lili ZHU ; Liming SUN ; Dongmei LI ; Beiwei ZHU
Acta Nutrimenta Sinica 2004;0(05):-
Objective To study the anti-tumor effect of the proteoglycan extracted from abalone viscera (AVPF-I). Methods Hepatic carcinoma cell H22-bearing mice were randomized to negative control (physiological saline), positive control (cyclophosphamide) and three doses of AVPF-I groups. The tumor growth, cell mediated immune function including lymphocyte proliferation, phagocytosis of peritoneal macrophage, NK cell activity, and the level of serum TNF-?,IL-1and IFN-?were detected. Results A significant inhibition of the tumor growth was observed in the AVPF-I group(P
2.Identification and activity assay of a polysaccharide from abalone harslet
Tingting CHENG ; Dongmei LI ; Na LIU ; Beiwei ZHU
Chinese Journal of Marine Drugs 1994;0(02):-
Objective To determine the purity,composition and antitumor activities of polysaccharide AHP-12 from abalone harslet.Methods Its purity was checked by HPLC with TSK-GEL G4000PWXL column and agarose gel electrophoresis;Molecular weight was determined by gel filtration chromatography;Sulfate content was identified by gelatine nephelometry and aminohexose content by chromatometry;Gas chromatograph was applied to determine monosaccharide composition;Antitumor activities were investigated by MTT method.Results AHP-12 from abalone harslet was a homogeneous polysaccharide both measured by molecular weight and electric property;The molecular weight was about 3?105;It was contained sulfate 13.07% and aminohexose 4.98%;AHP-12 was composed of rhamnose,fucose and galactose(the ratio in mole is 1∶2.2∶1.7);the activity determined by MTT method showed it could hamper the growth of HeLa cell.Conclusion AHP-12 was extracted and purified from abalone harslet.It was a homogeneous polysaccharide,which contained sulfate and aminopolysaccharide,and with weak antitumor activities in vitro.
3.Recombinant expression and antibacterial activity of i-type lysozyme from sea cucumber Stichopus japonicus.
Xiuxia WANG ; Lina CONG ; Dan WANG ; Xijian YANG ; Beiwei ZHU
Chinese Journal of Biotechnology 2009;25(2):189-194
The cDNA of an i type lysozyme was cloned from Stichopus japonicus (named as SjLys). The DNA fragment of the mature SjLys was subcloned into expression vector of pET-32a (+) to construct the recombinant plasmid of pET32a (+)-SjLys. The recombinant plasmid was then transformed into Escherichia coli BL21 (DE3) pLysS and induced by isopropylthio-beta-D-galactoside (IPTG). The recombinant protein expressed as inclusion bodies was denatured, partially purified and refolded to be an active form. The bacteriolytic activity of recombinant protein purified by the metal-chelating was 19.2 U/mg. The antibacterial activity of the purified recombinant SjLys (rSjLys) was analyzed. The rSjLys protein displayed inhibitive effect on the growth of the tested Gram-positive and Gram-negative bacteria. In particular, rSjLys had a strong inhibitive activity on Vibrio parahaemolyticus and Pseudomonas aeruginosa, both the most common pathogenic bacteria in the marine animals. The heat-treated rSjLys exhibited more potent activities against all tested bacteria. These results indicated that the S. japonicus lysozyme was the enzyme with combined enzymatic (glycosidase) and non-enzymatic antibacterial action, and it had a wide antibacterial spectrum. Therefore, it is suggested that the S. japonicus lysozyme should be one of the important molecules against pathogens in the innate immunity of sea cucumbers.
Animals
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Anti-Bacterial Agents
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metabolism
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pharmacology
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Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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genetics
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Muramidase
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biosynthesis
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genetics
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pharmacology
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Recombinant Proteins
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biosynthesis
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genetics
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pharmacology
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Stichopus
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enzymology