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Enzymatic Modification of Food Proteins (Characterizing hydrolysis ((3) Mw…
Enzymatic Modification of Food Proteins
Hydrolysis with
Proteases
Occurrence of protein hydrolysis
:
● Digestibility of proteins
● Fermented food products
● Hydrolysed food ingredients
Properties of protein hydrolysates
Nutritional
● Easier digestibility
● Loss of allergenicity
Bio-functional
● Anti-microbial properties
● ACE-inhibition
● Anti-oxidant activity
● Immunostimulation
● Antithrombotic Peptides
● Mineral Binding Peptides
Techno-functional
● Solubility
● Foam
● Emulsion
● Taste
Sources
:
Plant
● Papain (papaya fruits)
● Bromelain (pineapple)
Animal
● Pepsin; Rennin (stomach)
● Trypsin; Chymotrypsin (pancreas) ...
Microbes
● Bacteria
● Subtilisin (Bacillus species) ...
● Fungi
● Aspergillus oryzae
● Mucor miehei ...
Classification
:
● Based on
site of attack on the protein substrate
(endo-E or exo-E)
● Based on
catalytic amino acid residue in the active centre
of the protease molecule
(Serine protease - Aspartic protease - Cysteine protease - Metallo-protease)
Crosslinking with
Tyrosinase
/
Peroxidases
*
Crosslinking with
Transglutaminase
*
Chemical
(heat-induced)
● Crosslinking with lysino-alanyl formation
● Maillard reaction
Glycation
Crosslinking
Brown colour formation
● Acid hydrolysis
Characterizing hydrolysis
(3)
Mw distribution of peptides
● Possible link to techno-functionality (Foam, Solubility, Bitterness etc.)
● Descriptor for reproducibility of hydrolysis
● Size-exclusion chromatography (Reversed-phase chromatography)
(4)
Identification of peptides
● Presence of specific peptides, e.g. ACE inhibitors
● Determination of selectivity of proteases
● Reversed-phase chromatography (+ MS)
(2)
Concentration of intact protein
● Understanding the mechanism of hydrolysis
● (Zipper/One-by-one) Linderstrøm-Lang theory
● Size-exclusion chromatography
● Reversed-phase chromatography (+ MS)
(5)
Quantification of peptides
● Presence of specific peptides, e.g. ACE inhibitors
● Determination of selectivity of proteases
● Reversed-phase chromatography (+ MS)
(1)
Degree of Hydrolysis (DH)
● Overall extent of hydrolysis
● Kinetics of hydrolysis
● pH-stat (addition of HCl / NaOH)
● OPA/TNBS/Formol etc. (release of NH2 groups)