Isolation and Characterization of Protease Producing Bacterial from Soil and Estimation of Protease by Spectrophotometer

About 25 isolates were recovered from different soil samples collected from different field nearer to Ravulapalem village, East Godavari district, Andhra Pradesh, India. On the basis of colony size, texture, and microscopic characteristics, the isolates were categorized into 10types. Among these, five isolates showed good proteolysis activity, three isolates showed moderate activity, 2isolates showed poor activity, and eight isolates exhibited very poor activity. Structural, staining and biochemical activity results have revealed that four of five active enzyme producing bacteria are Bacillus sps, and one is staphylococcus sps. One Bacteria shown high Enzyme producing activity than the other. Estimation of enzyme producing activity of bacteria isolate was studied by Spectrophotometry. Crude enzyme extract was studied for protein degrade activity and producing amino acid amount  was measured by using folin phenol reagent and the absorbance was read at 660 nm using Spectrophotometer. The amount of protease produced was estimated and expressed in micro gram of tyrosine released under standard assay conditions. Based on the tyrosine released theprotease activity was expressed in micro gram of tyrosine released by 1 ml of enzyme in 30 minutes at 300C on tyrosine equivalent.