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Study on the Isolation, Identification, Bioactivity and Secondary Metabolites of Mangrove Actinomycetes

Author: LinCheng
Tutor: ShenYueMao
School: Xiamen University
Course: Microbiology
Keywords: Mangrove plants Within the supergene actinomycetes Biological activity Secondary metabolites
CLC: R284
Type: Master's thesis
Year: 2009
Downloads: 405
Quote: 2
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Abstract


The natural product is an important source of the drugs. Actinomycetes (Actinomycetes) has a unique advantage to produce antibiotics, about 75% of the antibiotics by actinomycetes produce secondary metabolites from endophytic actinomycetes isolated many novel structure, biological a variety of active compound. Mangrove plant growth in the marine intertidal give it a rich microbial resources, unique living environment is currently much emphasis on new medicinal resources. This thesis is born of Zhangzhou City, Fujian floating palace town of Jiulong River Estuary, Fujian Xiamen Jimei and Haicang three sites in two mangrove plants (candel, and Aegiceras), leaves and roots collected from within the table actinomycetes separation of bacteria, 55 actinomycetes. Directions of Bacillus subtilis (Bacillus subtilis), E. coli (Escherichia coli), Staphylococcus aureus (Staphlococcus aureus), Bacillus pumilus (Bacillus pumilus), Candida albicans (Candida albicans) and Aspergillus niger (Aspergillus niger) bacteria, using the double agar diffusion method for antibacterial activity of these strains, screened a total of 26 active strains, 47.3% of the total number of strains, which is the main activity of the anti-Gram-positive bacteria; MTT assay was used to its in vitro The anti-tumor activity detection, found a relatively high inhibition 5 actinomycetes Hela (uterine cancer) and HepG2 (human hepatoma cells), 9.1% of the total strain. By 16S rDNA sequence analysis, molecular identification of the isolates, and the results show that only a actinomycetes belonging intends Nocardia (Nocardiopsis), others are Streptomyces genus (streptomyces). AHBA primer isolates screened, 6 actinomycetes result is positive, indicating that the 6 bacteria DNA may contain AHBA (3 - Amino - 5 - hydroxy-benzoic acid) synthase gene, including contemplated Novo Cartesian bacteria. Secondary metabolites of this thesis, the strain intends Nocardia (No.3 bacteria) and has high anti-bacterial and anti-tumor activity of other strains of Streptomyces bacteria) (18 conducted a preliminary study. Isolated from the strain of secondary metabolites and identified 13 compounds, wherein three of the new compound (3C, 3Y-10) belong Pyrones; 10 to the known structure of the compound (3A, 3Y-5, 3Y-7, 3Y-14, 3L-A, 3L-M, 3L-G, 3Y-18, 3Y-19, 3Y-20): 3A, 3Y-5, 3Y-7 and 3Y-14 for derivative of tryptophan; 3L - A 3L-M type compound of the salicylamide; 3L-G is acetic acid glycerides; 3Y-18 is acetyl tyramine; 3Y-19, and 3Y anthranilic acid glycerides; -20 for the cyclic peptides. 4 of the known structure of the compound (18-C ,18-F ,18-E ,18-G) was isolated and identified from the 18 strains of secondary metabolites, respectively p-hydroxyphenyl acetic acid methyl ester, α- ketone dimethyl glutarate, and 2,6 - dideoxy-pyranoside, 2,6 - dideoxy-pyranose. Antibacterial activity and anti-tumor activity on the isolated compound 3C, shows no antimicrobial activity in each piece of filter paper containing 50μg sample (filter paper method) on five kinds of indicator bacteria; antitumor experimental results show, 3C Hela cells only slight inhibition in the two concentrations 10ug/mL and 20ug/mL when inhibition rate of 2.73% and 7.39%, respectively. Our results show that the mangrove actinomycetes bears rich anti-bacterial and anti-tumor substances producing bacteria, also bears a rich novel structure of the active substance, is the development of antimicrobial potential anti-tumor drugs such resources.

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