3-Aminothiophene-2-carboxylates 1-12 were screened for antimicrobial activities against gram positive bacteria Staphylococcus aureus (ATCC 29737) and gram negative bacteria Escherichia coli (ATCC 25922) and Candida albicans (MTCC 277) and Aspergillus niger (MCIM 545) fungi. Compounds 4, 5, 9containing chloro, methoxy and amide functionalities showed excellent to moderate antibacterial activities against Gram negative bacteria Escherichia coli and Gram positive bacteria Staphylococcus aureus with MIC 10-20μg/mL compared with standard antibiotic drug Gentamicin (10 μg/mL). Similarly, compounds5-7,8-12containing chloro, methoxy and amide groups showed excellent to moderate antifungal activities against Aspergillus niger and Candida albicans with MIC 10- 20μg/mL on comparison with standard drug Fluconazole (20 μg/mL).