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Accepted Manuscript

Title: Food peptides: Purification, identification and role in the


Author: Naima Arroume Renato Froidevaux Romain Kapel

Benoit Cudennec Rozenn Ravallec Christophe Flahaut
Laurent Bazinet Philippe Jacques Pascal Dhulster

PII: S2214-7993(16)30016-9
Reference: COFS 130

To appear in:

Received date: 8-1-2016

Revised date: 11-2-2016
Accepted date: 12-2-2016

Please cite this article as: Arroume, N., Froidevaux, R., Kapel, R., Cudennec,
B., Ravallec, R., Flahaut, C., Bazinet, L., Jacques, P., Dhulster, P.,Food
peptides: Purification, identification and role in the metabolism, COFS (2016),

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Food proteins as source to produce bioactive peptides

1. There is a high biodiversity of bioactive peptides from animal or plant sources

2. Bioactive peptides play an important role in energy homeostasis

3. Peptidome identification is needed to understand the bioactivity
4. Controlled bioprocesses are needed for the separation of bioactive peptides


Page 1 of 15
Food peptides: purification, identification
and role in the metabolism
Naima Arroume1, Rnato Froidevaux1, Romain Kapel1, Benoit Cudennec1, Rozenn
Ravallec1, Christophe Flahaut1, Laurent Bazinet2, Philippe Jacques1,3 and Pascal Dhulster1


Univ. Lille, EA 7394, USC 1281 - ICV- Institut Charles Viollette, F-59000 Lille, France

Institute of Nutrition and Functional Foods (INAF), Department of Food Sciences and
Laboratory of Food Processing and Electromembrane Processes (LTAPEM),Universit Laval,
Qubec (QC), G1V 0A6, Canada.

Terra Research Centre, Microbial Processes and Interactions, Gembloux Agro-Bio Tech,
University of Liege, Passage des Dports, B-5030 Gembloux, Belgium

Abstract an
The search for new active molecules from food protein hydrolysates represents a real
challenge for animal feed, aquaculture, as fertilizer, but also in cosmetics and
pharmacological fields. The constant discovery of molecules with biological activities
continually brings new insights into the complex mechanisms involved in vivo. However, it
remains difficult to conclude about the structure, amino acid composition, size or the

physicochemical properties necessary for the activity. This review will highlight the research
carried out to determine the physiological role of these peptides and the development of
new tools for their production and study.


The multiplicity of peptide sequences generated by the hydrolysis process provides a wide
range of structural and physicochemical properties that can help, using technological
processes such as ultrafiltration membranes, to separate them by family properties and to
draw the necessary conditions to maintain the activity [1]. There are yet no general rules for

determining the best hydrolysate with the most important activity. Each raw material, each
enzyme and each hydrolysis conditions will give different final peptides. The relationship
between the sequence and activity is still studied and represents a basic research for the
discovery of the complex mechanism of action of these biological activities [2,3,4,5]. A set of
recent reviews described the high biodiversity of bioactive peptides from animal and plant
proteins [6,7*,8**,9*,10]. In this short review, we will focus on the recent evolution of
several research fields related to protein hydrolysis such as the role of bioactive peptides in
energy homeostasis, the importance of peptidomics and the development of controlled
bioprocesses for the separation of bioactive peptides.

Page 2 of 15
Fig. 1. Summary of tools and type of studies on bioactive peptides

Dietary proteins as sources of bioactive peptides involved in energy homeostasis


In economically developed countries, eating behaviors, characterized by an increase in

energy intake which causes an increase in body mass, led to the metabolic syndrome. This
syndrome predisposes to many diseases such as type II diabetes, a deregulation of food

intake, hypertension, chronic inflammatory bowel disease (IBD) [11] and became one of the
most alarming public health problems of this century. Digestion has for a long time been

considered as a black box producing unit nutrient compounds such as amino acids involved
in metabolism and indispensable to the physiological function of the body. In recent years,
an abundance of studies have shown the involvement of peptides generated during the
digestion in many physiological functions [12] and especially in energy homeostasis and

consequently in the "metabolic syndrome" and in its various symptoms.

Regulation of food intake

During a meal, the presence of nutrients in the gastrointestinal tract results in the
production of signals that inform the brain about their qualities and quantities. Among these
signals, peptide hormones secreted by enteroendocrine cells such as cholecystokinins (CCK)
and glucagon-like peptide 1 (GLP-1) were particularly studied and are of great interest for
the development of new drugs and dietary supplements targeting obesity [13]. During
digestion, the products of hydrolysis of food proteins are known to have a high satiating
power, greater than that exercised by carbohydrates or lipids, stimulating the secretion of
gastrointestinal hormones [13]. Thus, recent work has identified peptides able to exert a

Page 3 of 15
strong stimulation on the secretion of hormones involved in the regulation of food intake
[14]. Another promising approach in the research for natural molecules that can be used in
functional foods concerns the identification of peptides exerting agonistic action to that of
CCK and GLP-1 by binding to their cellular receptors distributed in the gastrointestinal tract
and central nervous system. Thus, studies have identified peptides able to bind to these
receptors and to exert in vitro and in vivo agonistic action [15,16,17]. In this way, Pupovac
and Anderson [18] evidenced in rats that peptides arising from digestion contribute to
satiety by independent activation of both opioid and CCK-1 receptors.

Glucose metabolism

GLP-1, an intestinal incretin, well known to be involved in the food intake regulation, also
acts on the functionality of pancreatic islets mainly by stimulating insulin secretion [19] and

slowing the glucagon secretion. An interesting approach is to assume that peptides able to
cross the intestinal barrier will be also able to inhibit the action of the dipeptidyl peptidase

IV (DPP-IV), a multi-functional enzyme mainly involved in the degradation of GLP -1 and thus
consequently in the regulation of glucose metabolism. The incretin effect is drastically
reduced or lost in type 2 diabetes mellitus (T2DM). Lately, DPP-IV inhibitors have indeed
been considered as innovative molecules for the treatment of T2DM enhancing the GLP-1
activity and recovering the incretin effect. Therefore, incretin-based therapy using DPP-IV
drug inhibitors (gliptins) is one of the most recent alternative treatments of T2DM as lately
reviewed [20]. Nowadays, several works focus on the strategy to identify natural peptide
inhibitors of DPP-IV activity as an alternative to synthetic drugs. Such peptides could be
obtained by processing food (protein hydrolysis and microbial fermentation) or by
proteolysis occurring during digestion. Numerous studies had first reported that milk derived
products induced DPP-IV inhibition in vitro [21, 22]. Other studies evidenced DPP-IV

inhibitory peptides from different protein sources like rice bran hydrolysates [23], fishes or
macro-algae [24, 25]. A recent study demonstrated, in vitro, for the first time that a
cuttlefish hydrolysate submitted to gastrointestinal digestion could extend the incretin

effect via two mechanisms: the stimulation of intestinal GLP-1 and the inhibition of intestinal
DPP-IV activity [26*]. These results could be linked to those obtained with a porcine skin
gelatin hydrolysate, harboring a DPP-IV inhibition activity in vitro, and improved glucose

tolerance in diabetic rats in correlation with the increase of insulin and the GLP-1 plasma
levels, the inhibition of plasma DPP-IV activity and the decrease of the plasma glucagon level
[27]. Recently, new prospects demonstrated that the computer-driven screening of bioactive

peptides and the in silico/in vitro combinatory approaches are attractive tools for the more
rapid discovery of new bioactive peptides [28, 29].

The need to identify peptidomes to understand peptide bioactivities

Long time challenging, the identification of all peptides of a complex mixture has become
easier, rapid and accurate with the emergence of omics techniques [30**]. In the field of
bioactive peptides, peptidomics is by definition the analytical method of choice for the
structural elucidation of peptides and of their post-translational modifications. Peptidomics
is defined as the comprehensive characterization of peptides in a sample [31*]. Practically,
peptidomics is a several steps procedure. First, pre-purification methods are used to
separate peptides from all other molecules (carbohydrates, proteins, lipids, salts). Then a

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combination of peptide fractionation techniques leads to the separation of the different
peptides based on different physico-chemical properties (size-exclusion chromatography
(SEC), liquid chromatography (LC), capillary electrophoresis (CE), gel-free isoelectrofocusing
(GF-IEF)). Finally, Tandem mass spectrometry (matrix assisted laser desorption/ionization-
tandem mass spectrometry (MALDI-MS/MS), electrospray-mass spectrometry (ESI-MS)) and
bioinformatics are used to identify and quantify all peptides in the complex mixture [32, 33,
34**, 35, 36]. Today, the identification of peptides relies on the MS-based fragmentation
methods (collision induced dissociation (CID), electron collision or transfer dissociation (ECD
and ETD, respectively)), the data acquisition speed and the accuracy of the most recent mass

spectrometers [37*]. Moreover, the MS-data acquire way (data dependent acquisition (DDA)

or data independent acquisition (DIA); positive or negative mode; ion-mobility), the
repetition of biological and technical experiments, the bioinformatics tools used to match
the experimental MS-data and the protein databases used have a direct impact on the

exhaustive characterization of peptides [38]. Although the MS-based quantification methods
(label-free, isobaric and isotope labeling) are available and validated [34], the relative

quantification of bioactive peptides remains rare and therefore the batch-to-batch
reproducibility is too less often reported [39]. Software (owned MS-manufacturer, software
developed by academic laboratories, free-of-charge or costly) of identification and
quantification of peptides are numerous, and all having advantages and drawbacks that
invite the user to integrate the bioinformatics results in a metascoring system to obtain a
comprehensive view of the peptide heterogeneity of a sample. Even if peptidomics is well
adapted some limitations exist such as ranging from the small peptides (< 5 amino acids)
that are challenging to identifying by tandem-MS due to the low number of MS fragments
obtained [40]. The hydrophobic and hydrophilic nature of peptides also complexifies their
separation and requires the use of several methods of peptide fractionation. To end with,
databases and bioinformatics tools need to be upgraded to limit the false positive rate that

is 100 times higher in peptidomics than for proteomics [41].

Controlled separation and purification of bioactive peptides


Bioactive peptides produced by enzymatic proteolysis are in complex hydrolysates that

contain more than a hundred different peptides. As a consequence, the level of bioactivity of

hydrolysates containing bioactive peptides is most often low. A classical way to improve this
is to carry out separation processes for enriching the starting mixtures into bioactive
peptides [42*]. Several approaches can be adopted, based on different types of

chromatography, electrophoresis and membrane separation techniques, in relation with

hydrophobic/hydrophilic, charge, molecular size or affinity properties of peptides. Adoui et
al [43*] have developed a simple and non-expensive method to recover the active peptide
from complex bovine casein hydrolyzate by optimizing the separation conditions through
ionic strength and pH parameters. This method was used to separate antimicrobial peptides
from complex peptic hydrolysate of casein and of RuBisCO [44*]. In this last case, three new
pure antibacterial peptides were obtained in these hydrolysis conditions. Processes have
been also developed and are summarized hereafter. We have chosen to present some
innovations based on surfactant and charge-size combining properties, such as membrane
filtration-electrodialysis or foaming-draining processes.

Membrane-based processes

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Membrane filtration such as ultrafiltration [45] or nanofiltration [46] has been widely used
to do so and a recent review on the subject is available [47, 48, 49].. However, the choice of
membrane, implementation mode and operating condition was rather empirical (non
specific to the peptide of interest and the starting hydrolysate properties). To tackle this
issue, an interesting work proposed to base the choice of membrane molecular weight cut-
off on overall peptide retentions (between 0.4 and 0.7) [50]. These authors also proposed a
calibration method of UF membrane based on size-exclusion chromatography (SEC) analysis
of permeate and retentate that relates peptide retentions to their molecular weights.

Recently, an original methodology for predicting yield and enrichment of a complex

hydrolysate in a bioactive peptide has been proposed. This method couples (i) SEC
chromatogram of the hydrolysate and targeted peptide molecular weight to mass balance
equations and membrane calibration [51, 52]. The main problem related to classical

membrane processes lies on their poor selectivity mainly based on solute hydrodynamic
radius. Interestingly, Bargeman et al. [53, 54] and Lapointe et al. [55] designed a filtration

module that uses one ultrafiltration membrane and an electrical field as a driving force. The
electrical field strength in the feed solution was maximized in relation to convection in order
to obtain a selective separation of the charged target compounds. However, in these
electromembrane configurations, the selectivity and purity of peptides are limited by
pressure application and membrane fouling. For these reasons, a technology called
electrodialysis with filtration membrane (EDFM) was developed and carried out successfully
for the production of bioactive peptides (anti-hypertensive, anti-cancer, anti-diabetic or anti-
microbial) from various protein hydrolysates [45, 47, 56, 57]. EDFM couples size exclusion
capabilities of porous membranes (NF, UF and MF) with the charge selectivity of
electrodialysis (ED) and no pressure is applied in the ED cell. The potential applications of
EDFM were enlarged by carrying-out simultaneously the hydrolysis and the fractionation of

the generated bioactive peptides [58] and also by stacking filtration membranes of different
molecular weight cut-offs [56].

Extraction processes

Fractionation processes have already been used for fractionation of bioactive peptides, such

as antioxidant [59], antihypertensive [60], DPP-IV inhibitory peptides [61], ACE Inhibitory
peptides [62]. Among them, separation processes have been carried out in taking account
hydrophobicity property of peptides. Indeed, researches have been carried out on the

selective separation of opioid or antimicrobial peptides through ion-pairing assisted

extraction in water/organic solvent biphasic systems. Ion pair formation in an aqueous
phase, thanks to the addition of alkyl sulfonates or Sodium Dodecyl Sulfate agents, allowed
to selectively extracting these bioactive peptides in organic phases consisting of
dichloromethane or octan-1-ol [63]. Such extraction system was intensified using an
ultrafiltration membrane which stabilized the interface and provided a greater exchange
surface [64]. Other authors have worked on the combination of a membrane contactor and
an organic interface consisting of octan-1-ol for the extraction of opioid peptides [65].
Another way to intensify a solvent extraction system is to implement it in centrifugal
partition chromatography (CPC). Recently, a mixed ion mode that uses a charged molecule
to displace equilibrium in the two liquid phases demonstrated very promising results for

Page 6 of 15
purifying the antihypertensive peptide Val-Trp from the alfalfa white proteins hydrolysate
(purification factor around 40 in a single step) [66].

Foaming-Draining processes

Selective separation studies of protein through the exploitation of their surfactant

properties are numerous [67,68] but works on selective extraction of bioactive peptides are
very rare. First study is demonstrated by Dordhain et al. [69*]. They have isolated
antibacterial peptides in a foaming-draining process in exploiting differences in surfactant

properties of the peptides present in the hydrolysate, related to differences in

hydrophilicity, hydrophobicity and molecular size. Experimental designs enabled highlighting
the major influent parameters and improving enrichment of the bioactive peptide fraction.

Development prospects

Despite the numerous successes of the production and commercialization of hydrolysates
containing one or more bioactive peptides, many industrial obstacles limit the sales of
nutraceutical or functional products. One of the main obstacles is the regulation aspect and
the application for a health claim authorization.
In addition, the characterization of the biological effect of the dietary peptides needs to be
improved. The development of new in vitro assay is of great importance to mimic the
physiological environment and get closer to the in vivo effect of the bioactive peptides. The
always growing number of studies on the human gastro intestinal digestion will help to
generate enough knowledge to understand the becoming of the dietary proteins and the
target of the biological active peptides generated during the enzymatic process [70]. This
integrated approach which confronts the biochemistry aspect of the food proteins and the

physiological aspect of the gut will permit to improve the efficacy of the bioactive peptides
in human and to give added value to food protein hydrolysates.
In this context of better efficacy of the peptides activities and a better knowledge of their

targets, the substitution of drugs by active peptides able to fight against specific diseases
such as T2D or hypertension will be a safely and economical way to decrease the harmful
side-effects of treatment usually induced by drugs, to potentiate the drug effect and to

upgrade food processing by-products.

Recently, a number of computational methods have been successfully applied in predicting
the potential of proteins as precursors of novel bioactive sequences. Two main approaches

are now reported: the former uses in silico enzymatic digestion and the bioactivity
prediction using a combination of sequence biochemical properties and databases of known
bioactive peptides; the latter is based on structure-based molecular modelling approach
coupled to a docking study (if the 3D structure of the target protein is known) to evaluate
the binding affinity between bioactive peptides and their target [71]. In the future, the
mixing of in silico, in vitro and in vivo approaches will constitute without doubt the road to

In order to extend the industrial growth perspectives, it may be interesting to develop

innovative separating processes of active fractions before the hydrolysis phase.
Furthermore, the active peptide fraction selectivity or even its yield could be improved by
combining separative processes or in association with the hydrolysis reaction.

Page 7 of 15
On the other hand, the valorization issue of natural resources or co-products from food
industry has an interest only if fraction of the co-products is enhanced in terms of its
bioactive properties. Nonetheless, the hydrolysate fraction containing several bioactive
peptides could show a higher effect on animal and human nutrition/health, as the
bioavailability of the peptides into a mixture is enhanced compared to isolated peptides.
Using of hydrolysates in aquaculture allows improving the survival rate of animals and could
be a new method to promote this technology. The antimicrobial properties of fractions or of
purified peptides, revealed through enzyme hydrolysis, constitute another means to
enhance hydrolysates in food safety, animal health and in cosmetology.

In case of proteins shortage, the use of more exotic, less conventional proteins should be
considered such as the transformation of insects.

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This article is a good example of an approach to elucidate the structureactivity

relationship of a bioactive property (antimicrobial),and peptide structural properties and
determines the minimum antimicrobial peptide sequence of the haemoglobin alpha chain

[43*] Adoui F, Boughrera F, Chataigne G, Chihib NE, Dhulster P, Zidoune MN, Nedjar-
Arroume N: A simple method to separate the antimicrobial peptides from complex peptic

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casein hydrolysate and identification of a novel antibacterial domains within the sequence
of bovine s-casein. Int Rev Chem Eng 2013, 5:179-187.

This interesting article develops a simple and non-expansive method to recover the active
peptide from complex bovine casein hydrolyzate by optimizing the separation conditions
through ionic strength and pH parameters.

[44*] Kobbi S, Bougatef A, Balti R, Le flem G, Firdaous L, Bigan M, Chaabouni S, Dhulster P,

Nedjar N: Antibacterial Activity Of Novel Peptides Isolated From Protein Hydrolysates Of

RuBisCO Purified From Green Juice Alfalfa. J Funct Foods 2015, (accepted)

This article shows that three new antibacterial peptides from RuBisCO, don't show hemolytic
activity towards bovine erythrocytes and suggest that these peptides from RuBisCO could be

a beneficial ingredient for nutraceuticals.

[45] Pouliot Y, Wijers MC, Gauthier SF, Nadeau L: Fractionation of whey protein
hydrolyzates using charged UF/NF membranes. J Memb Sci 1999, 158: 105-114

[46] Fernandez A, Zhu Y, FitzGerald RJ, Riera, FA: Membrane fractionation of a -

lactoglobulin tryptic digest: effect of the membrane characteristics. J Chem Technol
Biotechnol 2014, 89: 508515
[47] Mosser M, Kapel R, Chevalot I, Olmos E, Blanchard F, Oriol E, Marc I, Marc A:
Fractionation of yeast extract by nanofiltration process to assess key compounds involved
in CHO culture improvement. Biotechnol progr. 2012 31 (4): 875-882.

[48] Bazinet L, Firdaous L: Separation of bioactive peptides by Membrane processes :

technologies and devices. Rec Patents Biotechnol 2013 7: 9-27.

[49] Muro C, Riera FA, Fernndez A : Advancements in the Fractionation of Milk

Biopeptides by Means of Membrane Processes, In Bioactive Food Peptides in Health and
Disease, edited by Blanca Hernandez-Ledesma and Chia-Chien Hsieh, InTech publisher, 2013

Chapter 10, pp.241-266.

[50] Chabeaud A, Vandanjon L, Bourseau P, Jaouen P, Gurard F : Fractionation by


ultrafiltration of a saithe protein hydrolysate (Pollachius virens): Effect of material and

molecular weight cut-off on the membrane performances, J Food Eng 2009, 91 (3), 408-

[51] Kapel R, Klingenberg F, Framboisier X, Dhulster P, Marc I: An original use of size

exclusion-HPLC for predicting the performances of batch ultrafiltration implemented to
enrich a complex protein hydrolysate in a targeted bioactive peptide. J membr sci 2011,
383 (1-2): 26-34

[52] Bodin A, Framboisier X, Alonso D, Marc I, Kapel R : Size-exclusion HPLC as a sensitive

and calibrationless method for complex peptide mixtures quantification. J Chrom B, In
press, 10.1016/j.jchromb.2015.09.035.

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[53] Bargeman G, Dohmen-Speelmans M, Recio I, Timmer M, Van Der Horst HC: Selective
isolation of cationic amino acids and peptides by electro-membrane filtration. Lait 2000,
80: 175-185

[54] Bargeman G, Koops GH, Houwing G, Breebaart I, Van Der Horst HC, Wessling M: The
development of electro-membrane filtration for the isolation of bioactive peptides: the
effect of membrane selection and operating parameters on the transport rate.
Desalination 2002, 149: 369-374

[55] Lapointe JF, Gauthier SF, Pouliot Y, Bouchard C: Selective separation of cationic
peptides from a tryptic hydrolysate of -lactoglobulin by electrofiltration. Biotech Bioeng
2006, 94: 223-233.

[56] Doyen A, Udenigwe CC, Mitchell P, Marette A, Aluko RE, Bazinet L: Anti-diabetic and

antihypertensive activities of two flaxseed protein hydrolysate fractions revealed
following their simultaneous separation by electrodialysis with ultrafiltration membranes.
Food Chem 2014, 145: 66-76.
[57] Doyen A, Beaulieu L, Saucier L, Pouliot Y, Bazinet L: Demonstration of in vitro
anticancer properties of peptide fractions from a snow crab by-products hydrolysate after
separation by electrodialysis with ultrafiltration membranes. Sep Purif Technol 2011, 78 :
321- 329.

[58] Doyen A, Husson E, Bazinet L: Use of an electrodialytic reactor for the simultaneous -
lactoglobulin enzymatic hydrolysis and fractionation of generated bioactive peptides. Food

Chem 2013, 136: 1193-1202.

[59] Kleekayai T, Harnedy PA, OKeeffe MB, Poyarkov AA, CunhaNeves A, Suntornsuk W,
FitzGerald RJ: Extraction of antioxidant and ACE inhibitory peptides from Thai traditional

fermented shrimp pastes. Food Chem 2015, 176: 441447.

[60] Baltia R, Bougatef A, Sila A, Guillochon D, Dhulster P, Nedjar-Arroume N: Nine novel


angiotensin I-converting enzyme (ACE) inhibitory peptides from cuttlefish (Sepia

officinalis) muscle protein hydrolysates and antihypertensive effect of the potent active
peptide in spontaneously hypertensive rats. Food Chem 2015, 170: 519-525.

[61] Le Maux S, Nongonierma AB, Murray B, Kelly PM, FitzGerald RJ: Identification of short
peptide sequences in the nanofiltration permeate of a bioactive whey protein hydrolysate.
Food Res Intern 2015, 77: 534-539.

[62] Yousr M Howell N: Antioxidant and ACE Inhibitory Bioactive Peptides Purified from Egg
Yolk Proteins. Int J Mol Sci 2015, 16: 2916129178.

[63] Vanhoute M, Froidevaux R, Vanvlassenbroeck A, Lecouturier D, Dhulster P and

Guillochon D: Ion-pairing separation of bioactive peptides using an aqueous/octan-1-ol
micro-extraction system from bovine haemoglobin complex hydrolysates. J. Chromatogr.
B. 2009, 877: 1683-1688.

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[64] Dhulster P, Kapel R, Froidevaux R, Nedjar-Arroume N, Fertin-Bazus A, Choisnard L,
Guillochon D: Advancement in intermediate opioid peptide production in an enzymatic
membrane reactor assisted by solvent extraction. Desalination 2012 148 (1-3): 221-226.

[65] Balchen M, Reusaert L, Pedersen-Bjergaard S: Electromembrane extraction of peptides.

J. Chromatogr. A 2008, 1194: 143-149

[66] Boudesocque L, Kapel R, Paris C, Dhulster P, Marc I, Renault JH : Concentration and

selective fractionation of an antihypertensive peptide from an alfalfa white proteins

hydrolysate by mixed ion-exchange centrifugal partition chromatography. J Chrom B 2012,
905: 23-30.

[67] Wei L, Zhaoliang W, Yanji W, Rui L, Di H: Isolation of soy whey proteins from
isoflavones in the concentrated solution using foam fractionation. Sep. Purif. Technol.

2015, 149: 31-37

[68] Burghoff B: Foam fractionation applications. J. Biotechnol. 2012, 161: 126 137
[69*] Dhordain P, Bigan M, Vanhoute M, Pierlot C, Aubry JM, Dhulster P, Guillochon D,
Froidevaux R. Optimization of peptide separation from complex peptide mixture by
foaming-draining system. Sep. Sci. Tech. 2012, 47: 654-66219.

Its the first time that a paper describes selective separation of bioactive peptides in relation
with their interfacial property in using a simple, eco-friendly and low-cost foaming-draining


[70] Capriotti AL, Caruso G, Cavaliere C, Samperi R, Ventura S, Chiozzi RZ and Lagan A

Identification of potential bioactive peptides generated by simulated gastrointestinal

digestion of soybean seeds and soy milk proteins. J Food Composition Analysis 2015,

[71] Jao CL, Hung CC, Tung YS, Lin PY, Chen MC, Hsu KC: The development of bioactive
peptides from dietary proteins as a dipeptidyl peptidase IV inhibitor for the management

of type 2 diabetes. BioMedicine 2015, 5:14.

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