Casein hydrolysis in PEG-salt two phase system by Eudragit-chymotrypsin bioconjugate

Area of Science:

• Biocatalysis
• Enzyme Engineering
• Protein Chemistry

Background:

• Enzyme immobilization is crucial for biocatalyst reusability and process efficiency.
• Aqueous two-phase systems (ATPS) offer a promising environment for enzymatic reactions.
• Eudragit S-100 is a pH-sensitive polymer suitable for enzyme conjugation.

Purpose of the Study:

• To develop and evaluate a novel bioconjugate of alpha-chymotrypsin and Eudragit S-100 for casein hydrolysis.
• To optimize casein hydrolysis using the bioconjugate in an aqueous two-phase system.
• To assess the reusability and efficiency of the developed bioconjugate.

Main Methods:

• Conjugation of alpha-chymotrypsin to Eudragit S-100.
• Implementation of an aqueous two-phase system (polyethylene glycol/phosphate) for enzymatic hydrolysis.
• Optimization of the reaction by periodic replacement of the salt phase.
• Assessment of product yield and bioconjugate reusability.

Main Results:

• The alpha-chymotrypsin-Eudragit S-100 bioconjugate effectively catalyzed casein hydrolysis.
• Replacing the lower salt phase during the reaction significantly increased product yield.
• The bioconjugate demonstrated good stability and reusability for up to six cycles.

Conclusions:

• The developed bioconjugate and ATPS provide an efficient and reusable system for casein hydrolysis.
• Process optimization through phase replacement enhances product recovery.
• This approach offers a sustainable alternative for enzymatic protein modification.