Transcript Dry-cured ham as a source of bioactive peptides

Antioxidant and Antihypertensive Natural
Peptides in Dry-Cured Ham By-Products
Dr. Leticia Mora
Institute of Agrochemistry and Food Technology (IATA-CSIC)
Meat Industry By-products
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By-products represent a cost for the meat processing sector as well as an important
environmental problem.
Meat production sector is one of the most contaminants due to the high generation of:
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Waste water,
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air pollution,
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solid waste.
Waste water
Air pollution (CO2 emissions)
The efficient use of meat by-products is very important to reduce the environmental impact of this
sector as well as an economical source.
Meat Industry By-products
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Most of the solid waste in the meat industry is produced during slaughtering.
Live animal
Manure
Slaughtered
Meat
By-products
Non-edible
Intestinal
contents
Fertilizers

Edible
0
Pharmaceuticals
Insulin
Hide
Bone
Blood
Human food
Animal feed
Leather Soups
Industries are making a strong effort converting by-products into useful sources of edible and
non-edible products, producing valuable products and functional ingredients with added-value
and economic potential.
Meat Industry By-products
Dry-cured ham slicering process for its commercialization
Commercial
slices
Animal feed industries
Human food-related uses:
•
•
Household broths and soups.
Fried rinds as appetizer.
Solid waste
Bioactive peptides generation
Bioactive peptides
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Small peptides (2-20 amino acids).
Inactive within the sequence of the parent proteins.
When liberated by the action of enzymes can act as regulatory compounds.
Natural
Food processing
Gastrointestinal digestion
Under controlled enzymatic conditions
By-products
Dry-cured ham as a source of bioactive products
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Dry-cured ham is a high-quality product very well valorated by consumers around the world.
During the processing of dry-cured ham occurs an intense proteolysis responsible for its
characteristic flavour and texture that results in the generation of small peptides and free
amino acids.
Proteolysis
Intact protein
Naturally generated peptides
Why not potential
bioactive peptides??
Dry-cured ham as a source of bioactive products
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High content in salt (5-6%) of dry-cured ham makes it non-recommended for
consumers with CVDs.
Synthetic drugs cause side effects.
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Captopril
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Search for non-toxic and naturally generated
peptides to control the factors influencing CVDs
as alternative to synthetic drugs is of great
interest among researchers.
Enalapril
A clinical study to evaluate the effect of dry-cured ham ingestion on
CVDs in 13293 individuals resulted on no association between the
higher levels of consumption and the incidence of CVDs.
Medicina Clinica (2009) 133, 754-580
HYPOTESIS: Dry-cured ham as a source of bioactive peptides
Antihypertensive peptides in dry-cured ham
ACE I inhibitory peptides
role in the regulation of blood pressure.
ACE Inhibitory peptides
Angiotensin I
ACE I
Angiotensin II
Inhibitors of Potent
ACE activity
vasoconstrictor
reduce systolic blood pressure.
Increase of blood pressure
Antihypertensive peptides in dry-cured ham
In vivo test
In vitro test
DRY-CURED HAM
ANTIHYPERTENSIVE ACTIVITY
100
90
80
70
60
50
40
30
20
10
0
After 8h of ingestion:
Decrease in the SBP of
-8.3 ± 0.5 mmHg
0
-2
0
5
10
15
20
25
-4
SBP changes (mmHg)
ACE inhibitory activity (%)
ACE INHIBITORY ACTIVITY IN A DRYCURED HAM EXTRACT
-6
-8
0
1
2 ham
3 is a source
4
5
Dry-cured
Peptides extract (mg/mL)
of naturally generated
antihypertensive peptides.
6
-10
-12
-14
Control
Iberian Ham extract
-16
Time after administration (h)
Food Research International, (2015), 78, 71-78.
30
Extraction
Pretreatment
Fractionation
Identification
Quantitation
SIZE-EXCLUSION CHROMATOGRAPHY
Fractionation according
to the molecular mass
Extraction
Pretreatment
Fractionation
Identification
Quantitation
SIZE-EXCLUSION CHROMATOGRAPHY
In vitro ACE inhibitory assay
82% of ACE inhibitory activity
Extraction
Pretreatment
Fractionation
Identification
Quantitation
SIZE-EXCLUSION CHROMATOGRAPHY
In vivo antihypertensive assay
(2, 4, 6, 8, and 24h)
Extraction
Pretreatment
Fractionation
Identification
Quantitation
SIZE-EXCLUSION CHROMATOGRAPHY
In vivo antihypertensive assay
(2, 4, 6, 8, and 24h)
After 8h ingestion: reduction
of SBP in -38.0 ± 0.5 mmHg
Extraction
Pretreatment
Fractionation
Identification
Quantitation
HIGH-PERFORMANCE LIQUID
CHROMATOGRAPHY
Extraction
Pretreatment
Fractionation
Identification
Quantitation
Highest ACE
inhibitory activity
Extraction
Pretreatment
nLC-ESI-MS/MS
Q/ToF Mass spectrometer
Fractionation
Identification
Sequence
PAPPK
KAAAAP
AMNPP
IKLPP
AAPLAP
KPVAAP
KPGRP
PSNPP
IAGRP
KVLPG
TGLKP
AAATP
KAAAATP
IC50(mM)
199,58
19,79
304,50
193,90
14,38
12,37
67,08
192,27
25,94
265,44
51,57
100,00
25,64
Quantitation
Antihypertensive peptides in dry-cured ham
Peptide AAATP
IC50=100 mM
After 8h of ingestion:
Decrease in the SBP of -25.62 ± 4.5 mmHg
The presence of antihypertensive peptides in drycured ham could counteract the negative effect of
salt and contribute for a better possibility of
consumption.
J Proteomics, (2013), 78, 499-507.
Antioxidant peptides in dry-cured ham
Peptide SNAAC
IC50=75.2 mM
Cooking resistance and maintenance of
antioxidant activity
a
a
a
a
Simulated gastrointestinal digestion
a
a
b
Escudero et al (2013), Food Chemistry, 138, 1282-1288.
Mora et al (2016), Food Chemistry, (under review).
Natural antioxidant peptides in dry-cured ham by-products
Effect of cooking on the antioxidant capacity of natural peptides
Cooking increases the
antioxidant activity in all the
assayed antioxidant
methodologies.
Effect of simulated gastrointestinal digestion
In vitro digestion affected the
sequence of many of the natural
peptides, resulting in
differences in the antioxidant
capacity of digested broths.
Peptidomic aproach to identify those peptides responsible for the main differences
in antioxidant activity between types of cooking
Conclusion
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The intense proteolysis occurred during the dry-curing process is responsible for the generation
of small peptides, many of them showing antihypertensive or antioxidant activities.
The cooking process and in vitro gastrointestinal digestion of dry-cured ham by-products
positively affect the sequence of natural peptides and the observed antioxidant activity of broths.
The identification of those peptides responsible for main bioactivity is possible through the
development of novel approaches using peptidomics and modern bioinformatic tools.
Acknowledgments
This work has received funding from the European Unions’ Seventh Framework
Programme for research, technological development and demonstration:
Grant Agreement No. PCIG13-GA-2013-614281
Grant from Generalitat Valenciana in Spain
Grant No. GV/2015/138
THANK YOU
Dr. Leticia Mora
[email protected]