Bioactive peptides in foods

Bioactive peptides in foods Per Saris Department of Food and Environmental Sciences Viikki Biocenter Faculty of Agriculture and Forestry 9 th Annual ScanBalt Forum 2010 Healthy future September 22-24, 2010, Tallinn, Estonia Maatalous-metsä tieteellinen tiedekunta Faculty of Agriculture and Forestry www.helsinki.fi/yliopisto 23.11.2010 1

University of Helsinki Founded in Turku 1640, Helsinki in 1828 Main functions are research, teaching and societal interaction Two official languages, teaching provided also in English 40 000 degree students Staff 7 300, of which 4 700 are researchers and teachers Functions in four campuses in Helsinki and in 20 other places in Finland www.helsinki.fi/yliopisto 23.11.2010 2

Four campuses Maatalous-metsätieteellinen tiedekunta www.helsinki.fi/yliopisto 23.11.2010 3

Viikki Campus Maatalous-metsätieteellinen tiedekunta www.helsinki.fi/yliopisto 23.11.2010 4

Bioactive peptides in foods What is it about? www.helsinki.fi/yliopisto 23.11.2010 5

Bioactive peptides in foods Bioactive www.helsinki.fi/yliopisto 23.11.2010 6

Bioactive peptides in foods Bioactive a peptide that can can cause an biological effect, bad or good www.helsinki.fi/yliopisto 23.11.2010 7

Bioactive peptides in foods What is a peptide? www.helsinki.fi/yliopisto 23.11.2010 8

A peptide is Peptides (from the Greekπεπτός, "digested" from πέσσειν "to digest") are short polymers of amino acids linked by peptide bonds. They have the same chemical structure as proteins, but are shorter in length. Wikipedia www.helsinki.fi/yliopisto 23.11.2010 9

Peptide versus protein Conventions One convention is that those peptide chains that are short enough to be made synthetically from the constituent amino acids are called peptides rather than proteins Another convention places an informal dividing line at approximately 50 amino acids in length (some people claim shorter lengths). This definition is somewhat arbitrary. Long peptides, such as the amyloid beta peptide linked to Alzheimer's disease, can be considered proteins; and small proteins, such as insulin, can be considered peptides. www.helsinki.fi/yliopisto 23.11.2010 10

Bioactive peptides in foods What is foods? www.helsinki.fi/yliopisto 23.11.2010 11

What is foods? www.helsinki.fi/yliopisto 23.11.2010 12

Foods contain peptides The peptide profile changes in the food upon storage www.helsinki.fi/yliopisto 23.11.2010 13

Foods contain peptides The peptide profile of foods changes during digestion www.helsinki.fi/yliopisto 23.11.2010 14

Effects of food peptides Good effects Bad effects www.helsinki.fi/yliopisto 23.11.2010 15

Bioactive peptides act via the intestine S layer protein A of Lactobacillus acidophilus NCFM regulates immature dendritic cell and T cell functions. Konstantinov ym 2008. PNAS 105, 19474-19479 www.helsinki.fi/yliopisto 23.11.2010 16

Good effect of food peptides Peptides lowering blood pressure from Valio, Finland from Calpis, Japan www.helsinki.fi/yliopisto 23.11.2010 17

Peptides lowering blood pressure They contain Val-Pro-Pro and Ile-Pro-Pro (VPP and IPP) They are formed fromβ-casein by the proteolytic activity of Lactobacillus helveticus peptide drink placebo VPP and IPP inhibit ACE Time (months) Tuomolehto et al., 2004 www.helsinki.fi/yliopisto 23.11.2010 18

VPP and IPP inhibit ACE Angiotensiini I Angiotensiini II contraction verisuonia of supistava blood vessels ACE EC 3.4.15.1 Bradykiniini verisuonia rentouttava relaxation of blood vessels non-effective Tehottomat hajoamistuotteet degradation products ACE = angiotensin ACE is a protease and high activity leads to high blood pressure VPP and IPP inhibit ACE www.helsinki.fi/yliopisto 23.11.2010 19

Ways to release bioactive peptides from proteins Enzymes Chemical treatment Physical treatment www.helsinki.fi/yliopisto 23.11.2010 20

Utilization of databases for designed release Proteins IProClass Enzymes Expasy Enzyme MEROPS Database of bioactive peptides BIOPEP www.helsinki.fi/yliopisto 23.11.2010 21

Release of ACE-inhibiting peptides Termolysin digests when P1 = Leu, Val, Ile, Phe P2 P1 P1 P2 Proline oligopeptidase digests when P1 = Pro Together they release (in theory) ACEinhibiting peptides www.helsinki.fi/yliopisto 23.11.2010 22

Bad effects of bioactive peptides Celiacia from reactions to gluten derived from prolamins of wheat, rye or barley gluteeni on Terve suolinukka Healthy intestine off Tuhoutunut suolinukka KUVA: M A R K KU M Ä K I celiacic intestine Wieser & Koehler 2008 www.helsinki.fi/yliopisto 23.11.2010 23

The unhealthy structures of gluten contain much proline (P) and glutamine (Q) Peptidi Sekvenssi Alkuperä Jakso 33-mer LQLQPFPQPQLPYPQPQLPYPQPQLPYPQP QPF α-2 gliadiini 56-88 26-mer FLQPQQPFPQQPQQPYPQQPQQPFPQ γ-5 gliadiini 26-51 20-mer LQPQQPFPQQPQQPYPQQPQ γ-5 gliadiini 60-79 20-mer QQQQPPFSQQQQSPFSQQQQ Gluteniini 19-mer LGQQQPFPPQQPYPQPQPF A-gliadiini 31-49 17-mer QLQPFPQPELPYPQPQS A-gliadiini 57-73 15-mer VQGQGIIQPQQPAQL γ-gliadiini 15-mer QQPPFSQQQQQPLPQ Gluteniini 14-mer PQPQLPYPQPQLPY α-2 gliadiini 62-75 13-mer LGQQQPFPPQQPY A-gliadiini 31-43 www.helsinki.fi/yliopisto 23.11.2010 24 12-mer FSQPQQQFPQPQ γ-5 gliadiini 102-113

Less than 9-mers of gluten are not immunoreactive Hartmann et al. 2006 www.helsinki.fi/yliopisto 23.11.2010 25

Sour dough with rye malt 99.5 % of the prolaminin was degraded 160000 140000 90000 80000 120000 70000 100000 80000 60000 40000 20000 0 Rye meal CA 30 C LS 8 DSM 20451 GshR TMW 1.103 CA 37 C 60000 50000 40000 30000 20000 10000 0 522 879 695 550 374 319 Rye malt CA 30 C LS 8 DSM 20451 GshR TMW 1.103 CA 37 C Sour dough prolaminin mg/kg Sour dough with rye malt Loponen et al. J. Agric. Food Chem. 2009, 57: 746 753 www.helsinki.fi/yliopisto 26

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Bioactive peptide into food using starters/probiotics www.helsinki.fi/yliopisto 23.11.2010 28

30 % of human milk contains nisin producers www.helsinki.fi/yliopisto 23.11.2010 29

Others continued J Hum Lact 2008; 24; 311 Leonides Fernández, Susana Delgado, Helena Herrero, Antonio Maldonado and Juan M. Rodríguez The Bacteriocin Nisin, an Effective Agent for the Treatment of Staphylococcal Mastitis During Lactation www.helsinki.fi/yliopisto 23.11.2010 30

Soy yogurt was made containing bioactive peptide nisin (E234) www.helsinki.fi/yliopisto 23.11.2010 31

Bioactive peptides into food using genetic modification Starters and probiotics are modified to produce bioactive peptides or proteolytic activity Great potential!!!!!!!!!! www.helsinki.fi/yliopisto 23.11.2010 32

Shooting at long range www.helsinki.fi/yliopisto 23.11.2010 33

Shooting at close range www.helsinki.fi/yliopisto 23.11.2010 34

Shooting at close range Inhibition of foodborne pathogens by recombinant, pathogen-binding, bacteriocin- producing lactic acid bacteria L. monocytogenes (GFP) + L. lactis (CBD A500) or Lb. plantarum (CBD A500) LPxTG ss usp45 6xHis P45 CBD prtp CW-anchor The CBD surface display cassette. Cleavage and sortase-mediated covalent linkage to cell-wall www.helsinki.fi/yliopisto 23.11.2010 35

Survival of Listeria after 8 h incubation in PBST (ph 7) mixed with L. lactis strains. 10 9 10 8 10 7 10 6 Listeria + L. lactis Vector CBD A500 WT Nisin CBD A500 + Nisin www.helsinki.fi/yliopisto 23.11.2010 36

3.0 x 10 8 2.0 x 10 8 1.0 x 10 8 Listeria + Lc. lactis MG1363(CBD A500) Lb. plantarum ALC-01 Lb. plantarum ALC-01(CBD A500) Pediocin producer Pediocin producer + binding Listeria www.helsinki.fi/yliopisto 23.11.2010 37

Acknowledgements PhD Timo Takala PhD Hanan Abbas Hilmi PhD students Ruiqing Li Yurui Li Shanna Liu Kari Kylä-Nikkilä Collaborator: Prof. Martin Loessner, Switzerland Dr. Jussi Loponen for his figures www.helsinki.fi/yliopisto 23.11.2010 38