I’m currently working on a possible research to determine possible optimal conditions of alfa amylase from bacillus sub and SSF. I’ve seen that the most used method to determine activity is the DNS method, but we don’t have access to those reactives. We would like to determine activity using the iodine-starch reaction.

The most used method with this reaction I’ve seen is the Fuwa method. I’ve seen this one been mostly used for amylase grown in mediums or previously crystallized, but our amylase will come from the moisture or the SSF, so directly following Fuwa’s procedure will not be possible. Could someone explicitly tell me which steps are followed with the Fuwa method or tell me an article where they are mentioned clearly for a similar arrangement that the one we want to use?

The other method I’ve seen is the one mentioned in Bergmeyer’s method of enzymatic analysis, but that one uses phosphate buffer and HCl instead of acetate buffer and acetic acid. I’ve not seen many people used this method though. I wonder why.