Dahlia Plant Seed has been known as bio active homogeny content for anti microbe shown towards Escherichia coli aureus. This research is aimed to isolate the pure anti microbe from endophyte fungi fragmentation of Dahlia Plant Seed (Dahlia variabilis) and the structural determination of pure homogeny, as well as to conduct the pharmacological test found. In the first research step, it searched the optimum media as to product anti microbe from Endophyte fungi of Dahlia Plant Seed. This research used the fermentation media (Huang et al: 2007) varied the carbon source with peptone and ammonia nitrogen source. Screening was done with diffusion method in order to pathogenic microbe verily Escherchia coli, Staphylococus aureus, Aspergillus niger and Candida albicans. The extract concentration of impure endophyte fungi LBKRCC 41 for Fusarium sp 50 μl/disk showed that the small anti bacteria activity in the second media variance of Huang et al by watching in 5th , 10th , 15th and 20th days. Yet, the extract concentration of the same fungi for LBKURCC 43 (Sporothrix sp) was hard, and showed the big activity in the second day of in the same media variance. Media of Huang et al peptone variance gave the defend energy around 6 mm at E. coli and 8-9.5 mm at S. aureus. Media of Huang et al in ammonia variance gave the defend energy about 8-8,5mm at S. aureus. The hard Endophyte fungi of LBKURCC 43 Sporothrix sp 50 ul/disk gave the defend energy 20.65-23.3 mm at E. coli, 14.65-17.5 mm at S. aureus with peptone media variance of Huang et al (2007), for ammonia media variance of Huang et al (2007), it had the defend energy around 19.85-22.35mm at E .coli and 16.65-17.5 mm at S. aureus. This showed that the real difference and positive control applied. Yet, the both endophyte fungi did not show any activity to Aspergillus niger and Candida albicans. The second test result of metabolism with QC-MS was found out Xanthin and cathechin. The research result showed that the both endophyte fungi could be as the homogeny anti biotic source.