Comparative Study of in Situ and ex Situ Enzymatic Hydrolysis of Milk Protein and Separation of Bioactive Peptides in an Electromembrane Reactor

Abstract

Tryptic hydrolysis of whey protein isolate was performed simultaneously during (in situ) and before (ex situ) fractionation by electrodialysis with ultrafiltration membrane (EDUF) to obtain bioactive peptides. Peptide migration to anionic (ARC –) and cationic (CRC +) peptide recovery compartments was strongly dependent on the digestion strategy used. Indeed, peptide migration to the ARC – was observed to be higher with in situ digestion while peptide migration to the CRC + was higher in an ex situ digestion: a final peptide concentration of 103.10 ± 2.76 μg/mL was found in the CRC + (ex situ) while it was 49.65 ± 6.13 μg/mL in the ARC – (in situ). HPLC-MS studies showed 23 major peaks that were generated by tryptic digestion of whey protein isolate. Seven of these peptides migrated to the ARC – while nine and eight peptides migrated to the CRC + for ex situ and in situ digestions, respectively. Among them, different antihypertensive, antimicrobial, and hypocholesterolemic peptides were recovered depending on the recovery compartment such as IDALNENK and VYVEELKP­TPEGDLEI­LLQK in ARC – and ALPMHIR, TKIPAVFK, VLVLDTDY­KK, and VAGTWY in CRC + regardless of the mode of enzymatic hydrolysis.

Fouling prevention of peptides from a tryptic whey hydrolysate during electromembrane processes by use of monovalent ion permselective membranes

“…The BiPRO WPI was hydrolyzed with pancreatic bovine trypsin (reference number: T9201) purchased from Sigma -Aldrich (St.Louis, MO, USA). Then, the hydrolysate was heated at 80°C during 30 min in order to inactivate the enzyme and avoid further breakdown of peptides 21 and freezedried. The final WPH was composed of 89% peptides and 11% salt.…”

Electrodialytic Processes: Market Overview, Membrane Phenomena, Recent Developments and Sustainable Strategies

“…Interestingly, the lowest-MWCO cationic compartment contained inhibitor peptides and amino acids, while the bioactive peptides were concentrated in the highest-MWCO cationic compartment. To avoid the pre-hydrolysis of the protein before EDUF treatment, Doyen et al [ 241 ] and more recently Suwal et al [ 242 ] tested the simultaneous enzymatic hydrolysis and fractionation of generated bioactive peptide. They observed that the peptide sequence ALPMHIR, identified as lactokinin and known to exert an important antihypertensive effect, was recovered with an estimated 66% migration rate [ 241 ] and that the peptide migration rate was found to be affected by the mode of enzymatic hydrolysis and separation [ 242 ].…”

“…To avoid the pre-hydrolysis of the protein before EDUF treatment, Doyen et al [ 241 ] and more recently Suwal et al [ 242 ] tested the simultaneous enzymatic hydrolysis and fractionation of generated bioactive peptide. They observed that the peptide sequence ALPMHIR, identified as lactokinin and known to exert an important antihypertensive effect, was recovered with an estimated 66% migration rate [ 241 ] and that the peptide migration rate was found to be affected by the mode of enzymatic hydrolysis and separation [ 242 ]. Current research is exploring other ways to optimize hydrolysis before EDUF, e.g., by using high hydrostatic pressure [ 243 ].…”

Lactoferrin-Derived Peptides as a Control Strategy against Skinborne Staphylococcal Biofilms

“…Furthermore, cosmeceutical formulations with biomimetic and bioactive peptides are industrially developed and marketed for stimulating collagen and elastin synthesis and improving skin healing [ 42 ]. Furthermore, emerging technologies based on simultaneous in situ enzymatic hydrolysis and fractionation by electrodialysis with ultrafiltration membranes (EDUF) could make the production of AMPs efficient and feasible from generally considered safe proteins [ 43 ]. Therefore, although for the first time a lactoferrin hydrolysate has been proposed to control skin-borne staphylococci, the results of our work could be profitably exploited in the production of water-based products compatible with the skin environment.…”