Amylases are among the most important industrial enzymes and also have great significance in biotechnological studies. In this study effect of different concentrations of various carbohydrates and amino acids on the production of amylase enzyme by Rhizopus oryzae were studied. The activity of amylase enzyme was determined by cup plate assay method as described by Chaurasia et al. (2015, 2017b). Five carbohydrates, viz., Glucose, Fructose, Galactose, Sucrose and Starch were used in different concentrations to study their effect on the production of amylase enzyme. The amylase enzyme production was increased with the increase in the concentrations of carbohydrate. The higher concentration (i.e, 1.5%) of carbohydrates proved to be the most effective for the maximum production of amylase enzyme. Among the carbohydrates, Starch and Glucose were found to be most favourable for maximum production of amylase enzyme while Galactose proved to be poor in this respect. In control (without carbohydrate) no trace of amylase enzyme activity was detected. Six amino acids, viz., DL- Iso-Leucine, DL-Methionine, DL-b-Phenylalanine, L-Asparagine, L-Glutamic acid and L-Lysine were also used in different concentrations to study their effect on the production of amylase enzyme. Among the amino acids, L-Asparagine (at 5000 ppm concentration) was found to be the most effective for the maximum production of amylase enzyme. DL-Iso-Leucine, DL- b-Phenylalanine and L-Lysine proved to be unfavourable for amylase production in which no trace of amylase enzyme was detected. Therefore, these amino acids have not influenced the production of amylase enzyme.