Bioactive components in algae

Susan Løvstad HoldtPhD, Researcher at DTU Environment

Chairman of the Seaweed Network in DK

Annual Plant Biotech Denmark Meeting March 3-4 2011-

LIFE-KU

Basics about algae

Algae of the world-

and a world of algae

Algae utilisation

Bioactive components

Algal choice

Algal Biorefinery research

The Seaweed Network in Denmark

Outline

BasicsAlgaeAlgae are a large and diverse group of simple, typically autotrophicorganisms, ranging from unicellular to multicellular forms. The largest and most complex marine forms are called seaweeds. They are photosynthetic, like plants, and "simple" because they lack the many distinct organs found in land plants

Microalgae in fresh and marine water

Different unicellular green algae (from left to right): Chlorella, without flagella; Chlamydomonas, with flagella, and colonies of 4 cells of Scenedesmus. (Wegeberg og Felby, 2009)

Open-pond Test Facility at Ashkelon (Seambiotic)

Flat Panel Airlift Reactor in Stuttgart, Germany (Subitec)

Microalgae production designsLight, light and….light (CO2 and nutrients)

Pigments from algae

Astaxanthin –

8000 US$/kg

Haematococcus sp.

http://www.themagicisbac.com/bac-files/haematococcus.jpghttp://algae4oil.com/_borders/clip_image001_000.jpghttp://www.edwardtufte.com/bboard/images/0000c7-699.jpg

Highly antioxidant

Seaweed / macroalgaeSeaweed is a loose colloquial term encompassing macroscopic, multicellular, benthic marine algae. The term includes some members of the red, brown and green algae

Differentiated into:•Thallus: the algal body •Lamina/frond: a flattened structure that is somewhat leaf-like •Sorus: spore cluster •Holdfast: specialized basal structure providing attachment •Haptera:finger-like extensions of holdfast anchoring to benthic substrate

Basics

Figure 1

No roots-

just holdfasts and hapteraNutrient uptake takes place at the entire thalli

and there is no need for benthic substrateCultivate in suspension or on other substrates

Figure 2. (a) Spores from Palmaria palmata settled on vinylon string (2

mm in diam), (b) spores germinated in 3 weeks in nursery tanks with added nutrient and aeration and transferred to the field at this stage, (c) harvestable thalli after 4

months of field cultivation (a-c seeded and cultivated by Maeve Edwards). (d-e) Seeded string with Alaria esculenta coiled around culture rope and (f) Alaria after approximately 120

days culture at sea (Arbona and Molla 2006).

Basics

Figure 3. (a) “Storm toss”

Chondrus crispus (Irish moss) harvester from 1975 (Prince Edwards Island, Canada)

equipped with waders and basket to drag through the shallow water at the beach and (b) a typical

Irish moss handraker (Pringle and Mathieson 1986). (c) Painting

by Carl Locher (1882), Tangsamlere ved Hornbæk Strand (Seaweed collecters at Hornbæk beach, Denmark). (d) natural harvest of drifting populations of Furcellaria lumbricalis in Denmark getting loaded on trucks and sold to Litex A/S to extract the “Danish agar”

(e) Ascophyllum handraked in Ireland 2008 at low tide and bundled to a metric tonnes “climeen”

dragged up shore at high tide (wheel barrow upside down on top of the “climeen”), (f) where a lorry drives down to the shore at low tide and picks it up (pictures by Maeve Edwards).

A world of seaweed

H a r v e s t

Polyculture in embayments in Yellow Sea region, China J. FANG

China knows how!

A world of seaweed

Cultivation

Polyculture in embayments in Yellow Sea region, China; Google Earth

Seaweed production in Indonesia; travelling exhibition: Earth from Above

Boat

A world of seaweed

Figure

5. World seaweed production (without Asia: 12,600,000

metric tonnes; 99.7%) in marine and brackish water, 2006 (offshore; metric tonnes wet weight; FAO 2008).

Figure 4. Globally harvested (□) and cultivated seaweed (○) in offshore marine and brackish water from 1950-2006 (FAO 2008).

A world of seaweed

Russian Federation818 t Phaeophyceae spp (brown seaweed)France45 t Asparagopsis spp+Undaria spp (Wakame)Spain1 t species unspecified

Europe

World production without Asia: 42.000t

The commercial seaweed production worldwide accounts for 20

% of the total aquaculture production

Ireland

Palmaria palmata(dulse in english,”søl”

in Danish)

Århus

Saccharina latissima (former

Laminaria saccharina)

(”sukkertang”

in Danish)

Chondrus crispus (Irish moss)contains the valuable stabilisingagent carrageenan (E407) used in a wide range of product such as toothpaste, gelling agent for mar-malade, whipped crème etc.

Gelling purposes Soil enrichment

The seaweed can serve as fertilizer for crops if grounded into powder or made into nutrient rich extracts.

Snack, food, feed or health products

Pleasant taste, minerals and vitamin and bioactive compounds are just some of the reasons for the applications of seaweed in the kitchen, stall and health products

Reference: Havets dyr og planter

Seaweed utilisation

Energy: 11,000 MJ/tonnes dried seaweed

Table

1. Current and potential future uses of seaweeds (free after Indergaard and Jensen 1991).

Organic acids etc.Fine chemicals

CoalFodder/feed supplementEstersCarrageenan

ChemicalsFertilizer supplementAlcoholsAgar

GasVegetables/spicesMethaneAlginate

PyrolysisMechanical treatmentFermentationExtraction

Seaweed utilization

Variation and evolution

Storage Cell walls

Laminarin (β -1, 3 glucan) Alginate, fucans, cellulose

Floridian starch (amylopectin like glucan)

Agar, carrageenan, xylan, cellulose

Starch Mannane, ulvane, xylan, cellulose

Storage Cell walls

Laminarin (β -1, 3 glucan) Alginate, fucans, cellulose

Floridian starch (amylopectin like glucan)

Agar, carrageenan, xylan, cellulose

Starch Mannane, ulvane, xylan, cellulose

Variation in content

Holdt and Kraan, 2011

Sustainable Energy Ireland, 2009

Microalgae compositionHigh lipid content during stress

Plaza et al, 2009

Pigments/Carotenoids: astaxanthin, cantaxanthin, lutein, violaxanthin•Antioxidant activity, immunomodulation, and cancer prevention

Fatty acids: EPA, oleic, lineolenic, palmitic, palmmitoleic, and DHA fatty acid•Reduce risk of certain heart diseases, antioxidant, and anti-microbial activity

Proteins: Phycobiliproteins•Immunomodulation, and anti-cancer activity, hepatoprotective, anti-inflammatory, and antioxidant properties

Polysaccharides: Sulfated polysaccharides and insoluble fibers•Anti-viral, anti-tumor, antihyperlipidemia, and anti-coagulant, reduce total and LDL cholesterol

Vitamins/tocopherols (vitamin E): •Antioxidant activity

Phenolic and volatile compounds: •Antioxidant and anti-microbial activity

Microalgal bioactive components

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SPIRULINA

Beta CaroteneProduces Vitamin-A

Which is good for the eyes

GLAControls cholesterol

Improve skin tone

AntioxidantsSlow down the Ageing process

PhycocyaninStrengthens the immune system

CalciumFor healthy bones

And teeth

IronHelps in the formation

Of haemoglobin

Vitamin-B ComplexFor effective metabolism

Of Nutrients

ProteinsVital for growth and development

Functional microalgae

Recognized by WHO as oneof the best food supplements to combat malnutrition

Seaweed compositionSeaweed is known for its high content of polysaccharides, minerals and certain vitamins

Difficult to conclude on the contents of the different componentsas they vary with geography, environment, within populations and

season

Proteins: Generally low content: 5-15% of dry weightGreen and red: 10-30%Palmaria and Porphyra (red): up to 47%

Lipids: up to 4%, rich in the omega 3-fatty acids

Polysaccharides: 35-60%

Minerals: Na, K, P, Ca, Mg, Fe, I

Vitamins: Vitamin A, B1

, B2

, B6

, B12

, C, D og E

(Murata and Nakazoe 2001)

Minerals & vitaminsSeaweed contain more minerals than any other food. This is mainly dueto the the surface cell wall polysaccharides that freely and selectivelyabsorb inorganic nutrients from the sea.This also include undesirable compounds….. May work as biofilter.

•seaweed contain all the minerals human needs including trace metals

Seaweed…

more than just a fertilizer

Craigie, 2010

Due to the presence of:Classic growth hormones: auxin, cytokinin, gibberelins etc.Hormone-like plant growth regulators: Betaines, polyamines,

signal peptides etc.Cytokinin (isoprenoid and aromatic) and indole 3-acetic acid (IAA)…etc.

Proteins

35% is like soy beans

Seaweed as protein substitute of fish meal (Soler-Vila et al 2009): ~ 10% Porphyra in feed to rainbow trout:

-

no changes in growth performance-

enhanced pigmentation

~ 5% Ulva in tilapia fish feed (Sebahattin et al 2009): -

increased growth

-

better feed conversion ratio (FCR), -

better protein efficiency ratio (PER)

30% substitution i salmon feed:-

colour

-tasteContains:Phycobili proteins: antioxidative effect (Plaza et al 2008)Lectin: aggregate blood cells (Murata and Nakazoe 2001)

10-30 % DW

10-47 % DW

5-15 % DW

Soy beans: 35 %

LipidsLipids in seaweed can be divided into:

SterolsTri-, di-

og monoacylglycerols

Phospholipids

Fatty acids:n-3 Poly Unsaturated Fatty Acids (PUFA)(α–linolenic acid and

eicosapentaenoic acid (EPA))• other n-3 PUFA such as 18:4n-3, not present in other organisms

→ reduction of -cardiovascular diseases -cerebrovascular diseases (Plaza et al 2008)

→ active against-edema -inflammatories/erythema-blood flow (Khan et al 2007)

Rich in sterols, such as fucosterol (especially in brown algae (Fucus))

-possible reduction of blood cholesterol (Plaza et al 2008)-anti-inflammatory (inhibits infections; Plaza et al 2008)

PigmentsFucoxanthin: pigment/cartenoid from Fucus species (6%)

• Antioxidant (Le Tutour et al 1998)• UV-B defence (Heo and Jeon 2008)• Preventive effect on cerebrovascular diseases

(change in brain blood flow;

Plaza et al 2008)

• Increased metabolisme (Plaza et al 2008)•Anti-obesity•Possible up-regulation of UCP1 in BAT (brown adipose tissue)•2% lipids from Undaria reduce White AT (g/kg body weight)

of mice and rats (Maeda et al 2008)

Holdt and Kraan, 2011

Polysaccharides

Holdt, 2009

Phenols mainly in brown algaelocated in the outer membrane in physode-vesicles

Porphyra: (shinorine and pophyra-334 phenol)Sargassum: 2-3% og 6%Ascophyllum: 4-13% og 5 %Fucus: 2%Laminaria: 0.2% and 0.2-2.6%

Antioxidant og anti-Staphylococcus effekt (Zubia et al 2008) Other effects of phlorotannin:

antiherbivour (#1, #111)may form complexes with alginic acidthey may even be excreted to the surrounding mediaantioxidant (in vitro #8, ESR cellul no cytotoxicity on human fetal lung fibantidiabetes (ref in #8)radiation protection (ref in #8)anti-cancer (ref in #8)anti-HIV (ref in #8)anti-allergic (ref in #8)anti-plasmin inhibition (#10)photochemoprevention (#10)antiproliferative activities (#18)

Other…

Associated bacterial communities

halogenated furanones antifouling (Delsea pulcra)(#70)kahalalide F treatment of lung cancer, tumors and AIDS (Bryopsis)(#70)furanones anti-fouling (#147, #148)

Halogenated compounds, highly active

Seaweed extractsanti-fungicidal (ref in #30 and #69)

anti-ulcerative activity (ref in #30)anti-oxidation (#67)anti-bacterial (#69, #125, #127 and #165)

anti-fouling (#157, #164)

anti-pathogenic bacterial (#167 and #170)Anti-inflammatory (#61)

antifouling invertebrate larvae (#106, #107, #152-154)antifouling, bacteria (#143 and #152)antifouling, fungus (#152)anti-pathogenic (#107)antifouling, micro- and macroalgae (#153)

Undaria / Wakame in pasta•Antioxidant properties, due to the content of phenols,

lipid composition and fucoxanthins analysed•10 % addition did not change the fllavour of the pasta•n-3:n6 fatty acids relationship vas 1:3 in the seaweed pasta and

1:15 in the normal pasta•Heat from cooking did not destroy the fucoxanthin

(Prabhasankar et al 2009)

Undaria in synergy with fish oils in rats•Analysed the lipid concentration in liver and serum and

the enzymatic activity involved in the fatty acid metabolism of the liver

•Reduced concentration of tricylglyceroles in serum and liver•Seaweed(19%), fish oil (4%) or seaweed and fish oil in diet•Greatest reduction was with diet of both seaweed and fish oil

•Also a synergetic effect between seaweed and fish oil in the enzymatic activity

(Murata et al 2001)

High rate algal biomass production for food, feed,High rate algal biomass production for food, feed,biochemicals and biofuelsbiochemicals and biofuels

Algal Biorefinery

Selection of suitable Algal species

Algal cultivation optimisation

Production of food supplements from algal biomass

Production of biochemicals

Biofuels production (biohydrogen, biogas, bioethanol)

Technological, societal, environmental and economical assessment of sustainability

Demonstration

(alginates, β-carotene, omega-3 fatty acids)

(antioxidants, pigments, phenolic compounds)

Biofertilizer

Algal Biorefinery

CultivationMarin Centre of the Great Belt (MKS)

Funded by: Slagelse local authority and LAG Ministry of FoodPartners: Slagelse, GEMBA Seafood Consulting, Bisserup Fisk,DTU Food, Fishermen at Omø

Commercial utilization of organic seaweed for consumptionDFFE: Bælternes Fiskeriforening, GEMBA, DTU

The Danish Shellfish Centre in LimfjordenNew species-

including seaweed Fornyelsesfonden: DSC, DMU, DTU Environment etc.

High in added value productsHigh growth rate/yieldLow cultivation costs/manpower

-Soil enrichment and energy potential are low in priority, however possible in the waste of all species

Sustainable Energy Ireland, 2009

Criteria for species selection in DK

Micro MacroPolysaccharides Low (4-60 %) High (15 to 75 %)

Lipids High (up to 40 %) Low (max 4 %)

Proteins Similar (6-60 %) Similar (5-50 %)

Pigments Similar/but different types

Similar/but different types

Phenolics (flavonoids)

Similar

(up to 16 %)

Similar

(up to 14 % in brown sp.)

Phenolics/flavonoids/phlorotannins

Pigments alginatefish feed (omega-3 and protein)

Algal choice depends on end-product(s)

Landbased (tanks) and bioreactors

•

Controlled –

light

–

nutrients–

flow

–

fouling•

Safe

•

Harvest!•

Expensive (energy, nutrients)

•

Maintenance (man power)

Off-shore (lines/floating)•

No control on growth parameters

•

Higher risk•

Cheaper

•

No use of agricultural land

Macro and microalgal cultivation

Recommendations by the Seaweed Network in DK

White paper for aimed at guiding decision makers within research grants, business, local authorities and national politicians in Denmark

Steering committee

Some important point:Biomass or research within utilization first?Logistics no problem: follow the fish!Production including breeding (we need the biomass)Enzyme development for pretreatment in order to utilize the biomassClear guidelines of legislation in regard to non or “novel food”

according to EU and daily intake recommendationsFeed ingredient potential (organic feed)

Thank you!

The Seaweed Network in DKsusan@akvakultur.dk

www.akvakultur.dk

(”tangnettet)Information in Danish

Why eat seaweed?

Taste good: Different

taste and also

usedas flavour

enhancer

Healthy: Low

in calories, loow

in fat,high

in sugars

(but dietary

fibres),

antioxidants

High

content

of

vitamins and minerals

Health

effects

(scientifically): anti-cancer, anti-virus,lower

the

risk

of

cardiovascular

diseases

etc.

Easy: Dried

and long

shelf-life. Collect

yourself

Beautyful: Sprinkle as decoration

or

build

in….