The prime need of molecules which is used for multiple drug resistance pathogens, including bacteria and fungus particularly for mycobacteria. Synthesized the heterocyclic compound containing dipeptide (DP4) using solid phase peptide synthesis (SPPS). Characterized by mass spectroscopy and amino acid sequence analysis using LC-MS-MS. Antibacterial and antifungal evaluation has completed by zone of inhibition and minimum inhibitory concentration. The antimycobacterial evaluation has completed using percentage of reduction in relative light units (RLU) by Luciferase Reporter Phage (LPR) Assay. The compound was found to be extremely powerful inhibition against Bacillus subtillis and pseudomonas vulgaris at the concentration of 6.25 µg/ml as compared with standard at 25 µg/ml. The percentage of reduction in the relative light unit (RLU) of the compound DP4 was found to be 44.20% at the concentration of 100 µg/ml against clinical isolate of Mycobacterium Tuberculosis (streptomycin, isoniazid, rifampicin and ethamputol resistant) as compared with control.
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