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The Idenfication of Bacillus Subtilis Pda0426 and Research of Antimicrobial Metabolites

Author: LiYan
Tutor: LiuXinLi
School: Shandong Institute of Light Industry
Course: Biochemical Engineering
Keywords: Bacteriostatic Bacillus subtilis Active product Appraisal Active
CLC: R915
Type: Master's thesis
Year: 2011
Downloads: 46
Quote: 0
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Abstract


Antibacterial substances produced by microbial screening antibacterial important source of new drugs , a lot of raw materials for the development of new drugs . With a large number of drug-resistant strains , the development of new antibacterial drugs is particularly important . Antibacterial activity was isolated strongly inhibited Staphylococcus aureus the bacterial strains pda0426, and the morphology and molecular biology of this strain identification to determine pda0426 strain as a Bacillus subtilis named as Bacillus subtilis pda0426. As an indicator of the antibacterial activity of the fermentation broth , the fermentation conditions of the Bacillus subtilis pda0426 preliminary optimization , to determine the fermentation medium was composed of peptone , glucose , NaCl, optimal culture conditions : initial pH was 8.0 , the culture temperature speed of 180 r / min, 30 ° C, the incubation time for three days . While the recovery of components of the index, the fermentation broth crude separation method , the use of LH-20 dextran gel column chromatography and reverse phase column chromatography using a crude separation of the active product . Crude separation and purification of the active component of the fermentation broth and purification , to give the nine main component . 1H-NMR, 13C-NMR, 2D NMR data , and MS data were analyzed to determine the components of the structure 4-11-1,4-11-2, R6-5-13.14-1 and R7-1.2 .3-13.18-5,4-11-2 with R6-5-13.14-1 compound of the same kind , the are macrolide ,4 -11-1 ring dipeptide . High flux and the MTT assay, the study of the biological activity , mainly to determine the structure of compound tested strains The minimum inhibitory concentration (MIC) as well as 50 % inhibition concentration ( IC50 ) on tumor cells was measured . The results show that the compound 4-11-2 and the R6-5-13.14-1 against Staphylococcus aureus the MIC of 2.0μg/mL, and PC3 IC50 41.0μg/mL activity against fungi is not very clear , the fermentation broth in inhibition of Staphylococcus aureus , the main active ingredient . As the crude extract of the fermentation broth on the number of phytopathogenic fungi have some inhibitory activity , a compound having antifungal activity is still need to continue the separation .

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