Marine By-products: Raw Material usefor Functional Ingredient Development

Maria HayesMaria Hayes 11 & Tomas Lafarga& Tomas Lafarga1111 Food Biosciences, Teagasc Food Research Centre, Ashtown, DublinFood Biosciences, Teagasc Food Research Centre, Ashtown, Dublin 1515

Outline

• Introduction

• Why By-products could be used for ingredientproduction?

• What by-product ingredients can be generated?

• How these are generated? Processes

• Applications?

Changes to the EU Common FisheriesPolicy

• February 2013, the European Parliament approved apackage of major reform to CFP.

• Introduction of an Eco-Friendly system based onMaximum Sustainable Yields (MSY).

• By-products may be viewed as a valuable source ofbioactive compounds including Functional andTechnologically relevant peptides and carbohydrates.

The Marine Functional FoodResearch Initiative

Sources

Herrin

gM

ackere

l

Macroalgae

Shel

l and

leg

which

conta

in

mea

t

White

and

brown

crab

meat

By-p

rodu

ct/co

-

prod

uct

Fish fins

Fish scales

Fish heads

Cancer pagurus

Fish gelatin

Mic

roalg

ae

Algal bioactive ingredients

Antioxidants

Blood pressure

Cancer

Inflammation

Obesity

Diabetes

Mental health

Laminarin

Fucoidan

LIPIDS& small molecules(phlorotannins, carotenoids)

Development of Chitosan Bread:Food Safety

Inhibitory effects of chitosan bread against B. cereus and rope formation when directlyinoculated in control and chitosan-containing bread (A)

Bacillus cereus

Potential Hazards…

• May not be suitablefor persons withshellfish allergies

• Depends on thepurity of Chitosanand if all protein isremoved during theproduction process

Proteins For The Future

• Between 1995 and 2020, about 97.5 percent of theglobal population increase will be in developingcountries, by which time 84 percent of the world’speople (an estimated 6.3 billion) will be living indeveloping nations.

• Need to widen the choice of dietary proteins forconsumers

• Up-scaling and sustainable production and use ofproteins from alternative marine sources is required

10

GELATIN IN THE PAST AND PRESENT 1

2013

The Frenchman Papin reports on a cooking process in which hetried to obtain a jelly-like mixture from bones

Gelatine (Latin: gelatus = stiff, frozen). The word gelatin is first usedin Europe around 1700

The first patent in the adhesive sector including gelatin, is granted inEngland in 1754.

Profound discoveries by the English doctor Richard LeachMaddox lead to decisive breakthrough in photography.

This year is considered to be a milestone in modern gelatinmanufacture: emergence of small factories and large quantities ofgelatine could be manufactured.

The gelatin industry sees rapid technological developments and makes tremendous progress towards achievinghigh standards in the production of gelatine and the quality products

An international study initiated by the European Gelatine Association (GME) in 1999 confirmed once again thatraw material regulations and gelatine production processes guarantee maximum safety for consumers.

16821682

1700

1754

1871

1875

1950

2001

First studies and publications regarding generation of gelatine from fish materials1956

1 Source: Modified from All About Gelatin. The Gelatin Manufactures of Europe (GME).

Gelatin is derived mainly from bovineand pig skin (46% of worlds gelatinoutput).

Bones of pigs and cattle represent23% of world output.

Only 1% of world gelatin outputcomes from marine sources: skins,scales and bones of fish.

11

• Fish Gelatin releases aroma and shows a higher digestibility than animal gelatin.

• Lower gelling temperature: advantageous for certain uses such as in electronicaltrade or precipitation of emulsions.

• No risk of health-threatening outbreaks of bovine spongiform encephalopathy(BSE) and foot and mouth disease (FMD).

• Acceptable for Hindus and members of the Islamic and Jewish faiths.

• Most gelatine-allergic patients do not react to fish skin gelatin.

ADVANTAGES OF FISH SKIN COLLAGEN/GELATIN

ACTUAL SOURCES OF COLLAGEN ANDGELATIN

1 Source: Gelatine information, news, history and more. Gelatine Manufacturers Institute of America. Retrieved 2008-09-26.

12

Gelatin generation methods from Salmon coGelatin generation methods from Salmon co--product cutsproduct cuts

Gelatin generated with the NaCl method: (A) Salmon Bones, (B) Salmon Skin, (C) Salmon Heads and (D) Salmon Offal

Gelatin/Collagen from Fish By-Products

Underutilised Fish Species: PotentialUnderutilised Fish Species: Potentialraw material for peptide generationraw material for peptide generation

•• White fish includingWhite fish includingblue white andblue white andherringherring

•• Capros aperCapros aper(Boarfish)(Boarfish)

Blue whiting mince extract Boarfish Myofibrillar protein extract

Amino acid Content Concentration g/Kg extract powder Concentration g/Kg powderCysteic Acid 11.747 0.093

Methionine Sulfone 35.178Asp 78.602 2.439Thre 32.52 0.699Ser 29.596 0.672Glu 109.035 3.38Gly 29.523 0.999Ala 40.341 1.269Cys 5.565 1.496Val 38.39 0.826Met 0.496Ile 36.31 0.445

Leu 54.745 1.136NorLeucine 16400 16400

Tyr 24.538 0.649Phe 30.685 0.856His 23.023 0.563Lys 62.605 1.118

NH3 10.329 117.406Arg 47.991 2.152Pro 24.305 0.854

Amino acid content of blue whiting proteinmince extract and boarfish myofibrillar protein

**

*

* BCAA = Branched chain amino acids Hayes et al., 2013

CompanyName

Commercial Name Source Bioactivity

Senmie Ekisu Valtyron (sold as ingredient forsupplements or foods)

sardine muscle - dipeptide (Val-Tyr) ACE inhibitor + anti-hypertensive

NipponSupplement

Katsuabushi oligopeptide bonito -oligopeptide converted to bioactivetripeptide (Leu-Lys-Pro)

ACE inhibitor + anti-hypertensive

Natural Factors PeptACE bonito -oligopeptide converted to bioactivetripeptide (Leu-Lys-Pro)

ACE inhibitor + anti-hypertensive

Metagenics Vasotensin bonito -oligopeptide converted to bioactivetripeptide (Leu-Lys-Pro)

ACE inhibitor + anti-hypertensive

Calpis Ameal tri-peptide (Val-Pro-Pro) anti-hypertensive

Tokiwa Yakuhin Lapis Support dipeptide (Val-Tys) anti-hypertensive

Copalis Protizen white fish hydrolysate stress relief

Yalacta Stabilium200 atlantic fish autolysate Stress Relief

Biothalassol Fortidium Liquamen white fish hydrolysate (Molva Molva) stress relief, anti-oxidant,lowers GI

ProperNutrition SeaCure fish fillet hydrolysate IBS, Ulcerative colitis andCrohn's disease

Copalis Nutripeptin white fish hydrolysate lowers postpradial bloodglucose

Copalis Collagen HM hydrolysed fish collagen skin & cartilage regeneration

Copalis Prolastin hydrolysed fish skins promotes ligament regeneration+ anti-oxidant

PE

PT

IDE

SF

RO

MM

AR

INE

SO

UR

CE

S

Glycine Betaine & DMSP

• Glycine betaine and DMSP areosmolytic, zwitterionic compoundsfound in food, microalgae andseaweed.

• Formed by oxidation of bioactivecholine in mammals and helps tomaintain normal cell volume underosmotic stress.

• The beneficial effects of glycinebetaine relate to the maintenance ofnormal blood concentrations ofhomocysteine.

• In 2011, the Panel on DieteticProducts, Nutrition and Allergies (NDA)of the European Food Safety Authority(EFSA) established a cause and effectrelationship between consumption ofglycine betaine and normalhomocysteine metabolism and grantedit a health claim pursuant to Article 13of regulation (EC) No. 1924/2006.

Generation of Glycine betaine and DMSPGeneration of Glycine betaine and DMSPenriched seaweed extracts: Accelerated Solventenriched seaweed extracts: Accelerated Solvent

Extraction (ASE).Extraction (ASE).

ASE

Load cellLoad cell

Fill withFill with solvent(ssolvent(s))(0.5(0.5--1 min.)1 min.)

Static extractionStatic extraction(5 min.)(5 min.)

Flush with fresh solventFlush with fresh solvent(0.5 min.)(0.5 min.)

Extract readyExtract ready(total time, 12(total time, 12--14 min14 min.)

Heat and pressuriseHeat and pressurise(5 min.)(5 min.)

Load cellLoad cell

Fill withFill with solvent(ssolvent(s))(0.5(0.5--1 min.)1 min.)

Static extractionStatic extraction(5 min.)(5 min.)

Flush with fresh solventFlush with fresh solvent(0.5 min.)(0.5 min.)

Extract readyExtract ready(total time, 12(total time, 12--14 min14 min.)

Heat and pressuriseHeat and pressurise(5 min.)(5 min.)

Purge with nitrogenPurge with nitrogen(1(1--2 min.)2 min.)

Generation of Glycine betaine and DMSPGeneration of Glycine betaine and DMSPenriched seaweed extracts: Accelerated Solventenriched seaweed extracts: Accelerated Solvent

Extraction (ASE).Extraction (ASE).

Location and date of collection of seaweed samplesanalysed in this study.

Generation of Glycine betaine and DMSPGeneration of Glycine betaine and DMSPenriched seaweed extracts: Accelerated Solventenriched seaweed extracts: Accelerated Solvent

Extraction (ASE).Extraction (ASE).

• Extraction of polar metabolites DMSP and Glycinebetaine was carried out using the DIONEX ASE 200system and water with pressure and temperature of 120bars & 50˚C; 3 cycles of 5 minutes duration.

• Recovered ASE(R) fractions were centrifuged andaliquots dried under nitrogen and subsequently freezedried prior to 1H-NMR analysis.

0.51.01.52.02.53.03.54.04.55.05.5f2 (ppm)

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

NutraMara samplesHSQCGP

Using 1H-NMR for metaboliteidentification

3-(trimethylsilyl)propionic acid-d4 (TSP) sodium salt was used as an internal standard and resonanceidentification of samples was performed by comparison of reference compounds acquired under similar

conditions.

500.162 MHz300 K

5 mm PABBOBroad Band

Observe probe

Using 1H-NMR for metaboliteidentification

Acquired Spectra were Line broadened manually phased and Baseline corrected using MestReNova v6.0.2Baseline correction was carried out using Whittaker smoother algorithm.

Estimated quantities of glycine betaine and DMSP found indifferent macroalgae ASE® extracts in this study.

Bioactivities – Renin Inhibition

Renin inhibition by betaines extracted from Irish

seaweed extracts generated using ASE

0

50

100

A. nodosum

ISCG0365

Codium

fragile

ISCG0029

C. nodicaulis

ISCG0070

F. serratus

ISCG0299

Control

Potential applications of Betaineextracts…

Application in baked goodsto prevent loss of moisture

Extracts use in sportsperformance products

Cognitive functionapplications for pregnantwomen

*

*****

* Kilibwa, United States Patent US 6,217,930 B, April 2001** Pryor, J. L., et al., (2012) Effect of betaine supplementation on cycling

Sprint performance, Journal of the International Society of Sports Nutrition, 9, 12.*** Asia Pac J ClinNutr, 2013, 22, 2, 319-324.

FACTORS OF IMPORTANCE

Safety of ingredients generated

Compliance with labelling laws and EFSA regulationsfor Novel Foods and Nutritional claims

Thank you for your kind attention

Contact details:

maria.hayes@teagasc.ie

Tel: +353 (0) 1 8059957

Thomas.lafarga@teagasc.ie