The objective is to precipitate peptides without damaging them, so that they could be added as functional ingredients.
Is TCA precipitation a good approach?
dissolve protein by NaOH .1N then precipitate proteins by 0.1N HCl, then coagulation by water bath.
precipitate vegetable protein extract depend on isoelecteric point , therefore pH of precipitate protein different from vegetable type to type
for me, the best: TCA!!
Add 1vol of TCA stock to 4 volumes of protein sample. Incubate 10 min at 4°C. Spin at 12000 rpm, 10 min. Remove supernatant leaving protein pellet intact. Then wash pellet with 200μl cold acetone. Spin 12000 rpm, 5 min . Repeat wash whit acetone. then dry pellet to 50 ºC for 1 hour. and resuspend pellet in in desired buffer.
TCA precipitation is not food grade! Use a food grade compatible protocol right from the start if the project aim is a food ingredient product. Ethanol precipitation methods at industrial scale can be problematic, requiring special factories. If the aim is just proof of function, the preparation method is not so critical.
Any industrial extraction method must be cost-effective before food companies will be interested, no matter how functional the peptides. Otherwise you're looking at the nutraceutical market. Suggest you get a food ingredient formulator on board.
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