I would say that is a project.

Results will depend very strongly upon the preparation of the plant protein. You may want to produce a large amount of substrate and preserve (freeze) it. Or you may want to prepare a panel of substrates from various plant sources and various preparation protocols.

Result also will depend strongly upon the protease. Will you use a commercial product? Or one that you make from crude microbial supernatant? In any case I suggest you prepare a large lot to use for the entire study.

Also pay attention to the digestion conditions, especially salts and pH.

John Carter, Thank you for the response.

I am currently working on a project focused on producing bioactive peptides via enzymatic hydrolysis of plant proteins using non-commercial bacterial alkaline protease. I am in the process of selecting the most suitable protease and optimizing its production. Could you advise on the key characteristics I should prioritize when selecting a bacterial alkaline protease for this application, particularly regarding its pH and temperature, stability and activity?

You definitely want to choose an enzyme you can prepare reproducibly.

Other choices depend on your ultimate application (impact). Are you making food, dietary supplement, animal feed, or fertilizer? The choice will dictate the requirement for purity and reproducibility of your enzyme and total process. Scale-up is easier with enzymes that are flexible. But high-value products (like drugs) can justify expensive controls for scaling.

What is your substrate? This also will work best with a reproducible preparation, including salts and pH at least. Is it soluble or colloidal? Some enzymes only work on soluble substrates, others can effectively chew away at colloids. You may use a solid-liquid mixture if it it characterized.

I am not restriction to any one plant protein as substrate. I can choose any, in mind I am planning for rice bran protein or mushroom, or any nut (groundnut).

Three is a good number. Have you done any research into how other people typically prepare these commodities to generate protein-rich products? If you make them yourself, from raw commodities, be sure to allow time (etc. resources) to conduct your purification process a few times to get it worked out. If you buy simply them, then procure enough of a single lot of each to complete your work. Or choose one protein and procure three different lots to compare.

Have you looked into how will you assay for bioactive peptides? Again, if you're headed toward a product that will be used in food, feed, or pharmaceuticals, consider choosing a protease preparation that is already approved as GRAS, because you probably won't be able to remove it from the final product.

Thank you, John Carter, for the suggestions. I will definitely keep all this in mind.