Dr Leticia Mora Institute of Agrochemistry and Food Technology IATACSIC Meat Industry Byproducts Byproducts represent a cost for the meat processing sector as well as an important environmental ID: 909042
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Slide1
Antioxidant and Antihypertensive Natural Peptides in Dry-Cured Ham By-Products
Dr. Leticia Mora
Institute
of
Agrochemistry
and
Food
Technology
(IATA-CSIC)
Slide2Meat Industry By-products
By-products represent a cost for the meat processing sector as well as an important environmental problem.
T
he
efficient use of meat by-products is very important
to reduce the environmental impact of this sector as well as an economical source.
Meat production sector is one of the most contaminants due to the high generation of:
Waste water, air pollution, solid waste.
Waste
water
Air
pollution
(CO
2
emissions
)
Slide3Most of the solid waste in the meat industry is produced during slaughtering.
I
ndustries
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.
0Live animalManureSlaughteredMeat
By-products
Edible
Non-
edible
Intestinal
contents
Pharmaceuticals
Hide
Bone
Blood
Human
food
Animal
feed
Meat
Industry
By-products
Fertilizers
Insulin
Leather
Soups
Slide4Human
food-related
uses
:
Household
broths and soups.
Fried rinds as appetizer.Dry-cured ham slicering process for its commercialization
Solid waste
Meat
Industry By-products
Commercial
slices
Animal
feed
industries
Slide5Bioactive
peptides
generation
Food processing
Gastrointestinal digestion
Natural
Under controlled
enzymatic conditionsBy-productsSmall 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.
Bioactive peptides
Slide6During
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.Dry-cured
ham
is a high-quality
product very
well
valorated
by
consumers
around
the
world
.
Dry-cured
ham as a source of bioactive
products
Proteolysis
Intact
protein
Naturally
generated
peptides
Why
not
potential
bioactive
peptides
??
Slide7High
content in salt
(5-6%) of
dry-cured
ham
makes it non-recommended for consumers
with CVDs.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.
Dry-cured
ham as a source of bioactive
products
Synthetic
drugs
cause
side
effects
.
Captopril
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
Slide8ACE I
inhibitory
peptides
role
in
the
regulation of blood pressure.Angiotensin IAngiotensin II
ACE Inhibitory peptidesACE IPotent vasoconstrictorIncrease of blood pressure
Inhibitors
of ACE
activity
reduce
systolic
blood
pressure
.
Antihypertensive
peptides
in
dry-cured
ham
Slide9Food
Research International
, (2015),
78, 71-78.
ACE INHIBITORY ACTIVITY IN A DRY-CURED HAM EXTRACT
DRY-CURED HAM ANTIHYPERTENSIVE ACTIVITY
After
8h of ingestion: Decrease in the SBP of-8.3 ± 0.5 mmHgIn vitro testIn vivo test
Antihypertensive
peptides in dry-cured
ham
Dry-cured
ham
is
a
source
of
naturally
generated
antihypertensive
peptides
.
Slide10SIZE-EXCLUSION CHROMATOGRAPHY
Fractionation
according
to
the
molecular mass
Extraction
Pretreatment
Fractionation
Identification
Quantitation
Slide11SIZE-EXCLUSION CHROMATOGRAPHY
Extraction
Pretreatment
Fractionation
Identification
Quantitation
In vitro
ACE inhibitory assay
82% of ACE
inhibitory
activity
Slide12SIZE-EXCLUSION CHROMATOGRAPHY
Extraction
Pretreatment
Fractionation
Identification
Quantitation
In vivo
antihypertensive assay
(2, 4, 6, 8, and 24h)
Slide13SIZE-EXCLUSION CHROMATOGRAPHY
Extraction
Pretreatment
Fractionation
Identification
Quantitation
After
8h
ingestion
: reduction
of SBP in -38.0 ± 0.5 mmHg
In vivo
antihypertensive
assay
(2, 4, 6, 8, and 24h)
Slide14HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
Extraction
Pretreatment
Fractionation
Identification
Quantitation
Slide15Extraction
Pretreatment
Identification
Quantitation
Highest
ACE
inhibitory
activity
Fractionation
Slide16Extraction
Pretreatment
Fractionation
Identification
Quantitation
nLC
-ESI-MS/MS
Q/ToF
Mass spectrometer
Sequence
IC
50
(
mM
)
PAPPK
199,58
KAAAAP
19,79
AMNPP
304,50
IKLPP
193,90
AAPLAP
14,38
KPVAAP
12,37
KPGRP
67,08
PSNPP
192,27
IAGRP
25,94
KVLPG
265,44
TGLKP
51,57
AAATP
100,00
KAAAATP
25,64
Slide17Peptide AAATP
IC50=100 mM
After
8h of
ingestion
:
Decrease in the SBP of -25.62 ± 4.5 mmHg
The presence of antihypertensive peptides in dry-cured ham could
counteract
the negative
effect
of
salt
and
contribute
for
a
better
possibility
of
consumption
.
J
Proteomics,
(2013), 78
, 499-507.
Antihypertensive peptides in dry-cured
ham
Slide18Peptide
SNAACIC50=75.2 m
M
Escudero
et al (2013), Food Chemistry, 138, 1282-1288.
Mora et al (2016), Food Chemistry, (under review).
Cooking resistance and
maintenance of antioxidant activitySimulated gastrointestinal digestionAntioxidant peptides in dry-cured ham
a
a
a
a
a
b
a
Slide19Effect
of
cooking
on
the antioxidant capacity of natural peptidesEffect of simulated gastrointestinal digestion
Cooking increases the antioxidant activity in
all
the assayed
antioxidant methodologies.
In vitro
digestion
affected
the
sequence
of
many
of
the
natural
peptides
,
resulting in differences in
the antioxidant
capacity of digested broths
. Natural antioxidant
peptides in dry-cured
ham by-products
Slide20Peptidomic
aproach to identify those peptides responsible for the main differences in antioxidant activity between types of cooking
Slide21Slide22Conclusion
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.
Slide23This work has received funding from the European Unions’ Seventh Framework Programme
for research, technological development and demonstration:Grant
Agreement No. PCIG13-GA-2013-614281
HIGHVALFOOD
Grant
from Generalitat Valenciana in Spain Grant No. GV/2015/138
Acknowledgments
Slide24Dr. Leticia Mora
lemoso@iata.csic.es
THANK YOU
FOR YOUR ATTENTION