Burns Chronic wounds (Zn-polysaccharide) Skin infections

Preservatives (Ag-polysaccharide - proved) Acne (Zn-polysaccharide - proved) Skin soothing Hair treatments e.g. dandruff (Zn-polysaccharide)

Nutritional & gastrointestinal disorders Slow release of Zn

Applications of Metal ComplexesCOSMETICS

PHARMACEUTICS

NUTRACEUTICALS

Improves stability Bioavailability of metal

ion Decreases toxicity

Synergistic activity

•

Advantages

Why in Red Microalgae ?

* The polysaccharide has biological bioactivities

* The polysaccharide is an anion-exchanger

* The polysaccharide is resistant to a wide range of: pH’s & temperatures

* The polysaccharide is not degraded by hyluronidase & carbohydrolases

* Commercial algal production is feasible

Metal Complexes

Nutrient starvation

Physiology

ParticipantsBen Burion University

Prof. Shosh AradProf. Yuval GolanProf. Moshe GottliebDr. Ofer DubinskyDr. Roshan ShresthaDr. Li Shi-YangDr. Edward EtesholaDr. Miri LapidotDr. Lena PlesserDr. Oshrat Levy-OntmanDr. Kalpana ShresthaDr. Marina KeidanMrina WiscousanDorit van MoppesMia BalliIrit Tzabag

University of Maryland

Prof. V.N. VakhariaProf. Yoni Zohar

Soroka Medical SchoolProf. Ron DaganProf. Yacob WeinsteinProf. Mahmud Huliel

Industries

Cell wall formation

o Cell wall protein(s)

o Cell cycle

o Golgi involvement

o Cell wall inhibitors/mutants

o Carbon partitioning

o Sulfation

X 30,000 X 12,000

G

G

S

X 12,000

Cell wall modified DCBresistant mutants

2,6,Dichlorbenzonitrile

Cl Cl

CN

Cl Cl

CN

DCBDCB

Carbon Metabolism

Floridean Starch

Floridoside

14 CO2

FLORIDOSIDE Dynamic

Carbon Pool

Matusi et al. Appl Biochem Biotechnol. 104;13-22 (2003)

Irritation Prevention

Mixing Polysaccharide with antigen induce more Ab’s than injection of pure ovalbumin. The adjuvant effect is lower or similar to Freund’s complete adjuvant

Mice generated Antibodies when fed with transgenic

red microalgae expressing Ovalbumin or Hepatitis

Ovalbumin Hepatitis BTN

Feeding with transgenic algae generated relative higher titer of IgA (than IgG) as compared with injected Ab

Red Microalgae as Cell FactoriesIPV-VP2

Sufotransferase Erythropoietin

Proof of Concept

Immune Response

The Polysaccharide Act as an Adjuvant

Transgenic Red Algae

Constituent

(% of dry matter)

Carbohydrates 50-70

Protein 20-40

Fatty acids 6-10

Ash 7-13

Fibers

Soluble 5-8

Insoluble 15-27

Total 20-23

Palmitic (C16:0) 36

Palmitoleic (C16:1) 3.0

Stearic (C18:0) 0.9

Oleic (C18:1) 1.9

Linoleic (C18:2) 6.8

Arachidonic (C20:4) 17.8

EPA (C20:5- w3) 34

C 18.6

Niacin 4.3

B1 0.21Thiamine

B2 0.8

Riboflavin

B6 0.06

Pyridoxine

E 1.9

Zeaxanthin 0. 5%

Na 0.4

K 0.6

Ca 1.5

Se 10

Fe 110

Mg 1100

Al 50

Zn 50

Mo 2.0

I 250

Unique combinationFatty Acids Vitamins Minerals

Biomass Composition

Structure of phycocyanobilin (above) and phycoerythrobilin (below)attached by thioether

linkage to the apoprotein.

Visible absorption spectra of aqueous solutions of three dyes isolated from red

microalgae;a)red(pink);b)blue; c) purple.

Structure of phycocyanobilin (above) and phycoerythrobilin (below)attached by thioether

linkage to the apoprotein.

Visible absorption spectra of aqueous solutions of three dyes isolated from red

microalgae;a)red(pink);b)blue; c) purple.

PhycocyaninPhycoerythrin

Applications

Diagnostics

Cosmetics

Phycobiliproteins

A l g a l C o l o r s

Expression in AlgaeOversulfated alginate New types of highly sulfated agar, agarose, carrageenan etc.etc.

Expression in BacteriaOversulfated polysaccharides: xanthan gum, dextran, alginate alginate

Expression in PlantsSulfated cellulose – new type of wood and paperSulfated cotton – new fabricsSulfated polysaccharides resemble heparinProtection against fungi attack

Expression of ST in other organisms has a potential to generate new materials

Algal Sulfotransferase

16:0

18:0 18:1 18:2 18:3

18:3

desaturase

desaturase ElongaseElongase4

Desaturase

6

18:4

5

A l g a e

P l a n t s

9 Desaturase

12Desaturase

15 Desaturase

Engineering the pathway for production of EPA, AA and DHA in plants

AA

desaturase

Elongase6

desaturase6

20:3 20:4 20:5 22:5 22:6

DHA

5

EPA

SDAgLA

17desaturase

aLA

Plant-AL

Engineered PathwayProduction of EPA, AA & DHA in Plants

Control

Transgenic algae NuclearChloroplast

Transfer MethodsGlass Beads

Bombardment

Cell State

VectorsPromoters

Gene Transfer Into Algal Cells

Generation of anti-ovalbumin antibodies by feeding mice with transgenic algae

•••••••••••••••••

PROOF OF CONCEPT

Feeding withPure ovalbumin

Feeding withAlgal biomass

Mice were fed with algal biomass once a week (5 gr biomass powder)

O.D

. (4

50

nm

)

0.3

0.8

1.3

1.8

2.3

2.8

0 III V VII X

WTTransgenic

Weeks

0 III V VII

0.6

0.4

0.5

0.3

0.2

0.1

0

0.7

Weeks

Antibody generation after feeding

commercial ovalbumin

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

0 III V VII

No of feeding

O.D

. (6

30

nm

)

250 µg ova

2.5 mg ova

0 III V VII

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

O.D

. (4

50

nm

)

Weeks

Feeding with Transformed Algae

0

5000

10000

15000

20000

25000

1

Ant

ibod

y tit

ers

The polysaccharide act as an Adjuvant