Digestomica delle proteine: piattaforme integrate ... - .Digestomica delle proteine: piattaforme

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Digestomica delle proteine: piattaforme integrate per lidentificazione di peptidi

bioattivi e funzionali derivati dagli alimenti

Dr. Gianluca PicarielloIstituto di Scienze dellAlimentazione (ISA) - CNR, Avellino

picariello@isa.cnr.it

Bologna, 21 ottobre 2016

1913 !

food protein

food-derivedpeptides

inactive

active (or immunogenic??)

FermentationEnzymatic/acid hydrolysisRipening (proteolysis) Gastrointestinal digestion

MILK DERIVED BIOACTIVE PEPTIDES

Caseins (78-80%)(phosphoproteins)

s1-caseins2-casein-casein-casein

Whey proteins (18-20%)

-lactalbumin-lactoglobulinlactoferrin

Bioactive peptides

Milk-derived bioactive peptides

http://www.google.it/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=Xm190mom8Aq4kM&tbnid=1FRgscou8neNvM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S0924224401000036&ei=lWGDUvueBOHb0QXA9YHwDQ&bvm=bv.56343320,d.ZGU&psig=AFQjCNEbTJ3J4e2_pEFSNH4RF4hUOLMxbg&ust=1384428290374358http://www.google.it/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=Xm190mom8Aq4kM&tbnid=1FRgscou8neNvM:&ved=0CAUQjRw&url=http://www.sciencedirect.com/science/article/pii/S0924224401000036&ei=lWGDUvueBOHb0QXA9YHwDQ&bvm=bv.56343320,d.ZGU&psig=AFQjCNEbTJ3J4e2_pEFSNH4RF4hUOLMxbg&ust=1384428290374358

1984

2009

Food derived-bioactive peptides

Do they have a functional role

Non-protein components of digestion buffers affect digestion

Digest from the entire gliadin fractionfrom hexaploid wheat

Digest from whole casein and whey protein fractions

Complexity of the food matrix

2011-2015

Including an intestinal (jejunal)step of simulated digestion villi BBM

http://www.google.it/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&uact=8&ved=0CAcQjRw&url=http://apbrwww5.apsu.edu/thompsonj/Anatomy%20%26%20Physiology/2020/2020%20Exam%20Reviews/Exam%203/CH23%20Intestinal%20Histology.htm&ei=6Xj5VJfvIsbYPYmlgfgL&bvm=bv.87611401,d.ZWU&psig=AFQjCNFxss9K7ReZyjqFZ_Zl3Yq9vxeMYw&ust=1425721910939377http://www.google.it/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&uact=8&ved=0CAcQjRw&url=http://apbrwww5.apsu.edu/thompsonj/Anatomy%20%26%20Physiology/2020/2020%20Exam%20Reviews/Exam%203/CH23%20Intestinal%20Histology.htm&ei=6Xj5VJfvIsbYPYmlgfgL&bvm=bv.87611401,d.ZWU&psig=AFQjCNFxss9K7ReZyjqFZ_Zl3Yq9vxeMYw&ust=1425721910939377

VALIDATING THE DIGESTION MODELS

Ex vivo

In vivo

Absorption of bioactive peptides (bioavailability)

A few peptides (potentially immunogenic ratherthan bioactive) at very low amount detected in the basolateral chamber

Gastro-duodenal digests of milk proteinsapplied to the apical chamber

Case studies: milk, wheat, ancient cereals, meat, tree nuts

March 2016

Mature milk (2-4 weeks after term delivery) from 12 healthy donors:- Six mothers observing a strict milk (and dairy-products) free diet- Six mothers drinking a cup of bovine milk daily and eating dairy products

Peptide from BOVINE MILK Protein of origin (position)Uniprot

protein ID

Number of times

identified

Motherson milk-free diet

(N=6)

Milk-drinking mothers

(N=6)

YVEELKPTPEGDL -lactoglobulin (42-54) P02754 2 - 2

YVEELKPTPEGDLEIL -lactoglobulin (42-57) P02754 1 - 1

SDIPNPIGSENSEKTTM*PLW s1-casein (180-197) P02662 1 - 1

Aug 2016

Concluding remarks

Many supposed food-derived peptides are not stable to gastrointestinal digestion

Stability to digestion should be assessed to confirm a possible (local or systemic)activity

The identification of digestion stable peptides should be performed on real foodsystems, using physiologically realistic digestion/absorption models

The characterization of the digestome requires specialized and specifically designedanalytical methodologies

Analysis of the protein "digestome" : needs of integrated approaches

bioinformatics

food

fractionation

immunoreactivity

cell biology

perspective: metagenomics

CREDITS

ISA-CNR

Gianfranco Mamone

Giuseppe Iacomino

PhD student: Luigia Di Stasio

Simonetta Caira (now ISPAAM)

Director: Sabato DAuria

UNINA Federico II

Prof. Pasquale Ferranti

Prof. Francesco Addeo (retired)

Diapositiva numero 1Diapositiva numero 2Diapositiva numero 3Diapositiva numero 4Diapositiva numero 5Diapositiva numero 6Diapositiva numero 7Diapositiva numero 8Diapositiva numero 9Diapositiva numero 10Diapositiva numero 11Diapositiva numero 12Diapositiva numero 13Diapositiva numero 14Diapositiva numero 15Diapositiva numero 16Diapositiva numero 17Diapositiva numero 18