Biotechnology of algae in South Corea. Challenge in the present and in the future.

Marine Algal Biotechnology in Korea:

present and future perspective

Taejun Han, Prof, Dr

The Blue Economy

“The Blue Economy” ▷ Dr. Gunter Pauli

▷ Published in 2010

Ocean of the Earth

▷ 300,000 species

▷ 75% of the total O2 production

What is Blue Economy ?

The Red Economy

planned/based on what we do not have

The Blue Economy

invest less/innovate more /based on what we have

Ecosystem service

Service market

▷ Total $33,000 billion per year in 21st _century

▷ Marine ecosystem ($20,900 billion)

Biomonitoring of water quality

“sporulation”

The Ulva biotest

y = -409.9x + 98.551 r2 = 0.8809

0 20 40 60 80 100

0.0 0.2 0.4 0.6 0.8 1.0 Cu concentrations (mg/L)

S

p

o

ru

la

ti

o

n

(

%

)

image analyser

y = -407.86x + 99.688 r2 = 0.8975

0 20 40 60 80 100

0.0 0.2 0.4 0.6 0.8 1.0 Cu concentrations (mg/L)

S

p

o

ru

la

ti

o

n

(

%

)

visual

The Ulva biotest

Toxicants No. of samples

Metals 13 species

VOCs 7 species

Herbicides 3 species

individual compounds 3 species

sludge elutriates 9 species

slag leaching water 4 species

Oil 1 species

Wastewater 4 species

Stream water 13 species

Sea water 17 species

Totals 74 species

Heavy metals

Toxic chemicals

Waste Waters

Oils

TU Min Max CV(%)

Oil 10.6 8.8 18.1 9.74

Emulsifier 111.1 66.6 141.5 12.79

Oil + Emulsifier 534.6 455.2 655.1 7.57

0 100 200 300 400 500 600 700

Oil Emulsifier Oil + Emulsifier

T

o

x

ic

u

n

it

Set point

Mapping Data input

(Surfer 7.0 program) 126 126.1 126.2 126.3 126.4 126.5 126.6 126.7

37.2 37.3 37.4 37.5 37.6 37.7 37.8 1 2 3 4 6 7 8 10 11 12 13 14 17 18 23 24 °- È- µµ ÀΠõ½Ã ¿µÁ¾µµ ±³ µ¿µµ ¼®¸ ðµµ ½ÃÈ- ¹ æÁ¶Á¦ ´ ëº Î µµ ¿µÈï µµ ´ öÀûµµ

1µî ±Þ 2µî ±Þ 3µî ±Þ µî ±Þ¿Ü

¿¬¾È ¼öÁú 4º бâ COD

-0.2 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4

126 126.1 126.2 126.3 126.4 126.5 126.6 126.7 37.2 37.3 37.4 37.5 37.6 37.7 37.8 1 2 3 4 6 7 8 10 11 12 13 14 17 18 23 24 °- È- µµ ÀΠõ½Ã ¿µÁ¾µµ ±³ µ¿µµ ¼®¸ ðµµ ½ÃÈ- ¹ æÁ¶Á¦ ´ ëº Î µµ ¿µÈï µµ ´ öÀûµµ

1µî ±Þ 2µî ±Þ 3µî ±Þ µî ±Þ¿Ü

¿¬¾È ¼öÁú 4º бâ TN

-0.1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1

126 126.1 126.2 126.3 126.4 126.5 126.6 126.7 37.2 37.3 37.4 37.5 37.6 37.7 37.8 1 2 3 4 6 7 8 10 11 12 13 14 17 18 23 24 °- È- µµ ÀΠõ½Ã ¿µÁ¾µµ ±³ µ¿µµ ¼®¸ ðµµ ½ÃÈ- ¹ æÁ¶Á¦ ´ ëº Î µµ ¿µÈï µµ ´ öÀûµµ

1µî ±Þ 2µî ±Þ 3µî ±Þ µî ±Þ¿Ü

¿¬¾È ¼öÁú 4º бâ TP

-0.05 -0.02 0.01 0.04 0.07 0.1 0.13 0.16 0.19 0.22 0.25 0.28 0.31 0.34 0.37 0.4 0.43 126 126.1 126.2 126.3 126.4 126.5 126.6 126.7

37.2 37.3 37.4 37.5 37.6 37.7 37.8 1 2 3 4 6 7 8 10 11 12 13 14 17 18 23 24 °- È- µµ ÀΠõ½Ã ¿µÁ¾µµ ±³ µ¿µµ ¼®¸ ðµµ ½ÃÈ- ¹ æÁ¶Á¦ ´ ëº Î µµ ¿µÈï µµ ´ öÀûµµ

1 2 3 4 5

¿¬¾È ¼öÁú 4º бâ Ulva Index

-0.2 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8

Ulva index COD

T-N T-P

autumn winter spring 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 50 100 150 200 250 300 350 B O D ( m g · L -1) TU (S) BOD 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 20 40 60 80 100 120 140 160 C O D ( m g · L -1) TU (S) COD 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 10 20 30 40 50 60 70 T -N ( m g · L -1) TU (S) T-N 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 50 100 150 200 250 300 350 B O D ( m g · L -1) TU (S) BOD 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 20 40 60 80 100 120 140 160 C O D ( m g · L -1) TU (S) COD 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 10 20 30 40 50 60 70 T -N ( m g · L -1) TU (S) T-N 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Month TU 0 50 100 150 200 250 300 350 B O D ( m g · L -1) TU BOD 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Month TU 0 20 40 60 80 100 120 140 160 C O D ( m g · L -1) TU COD 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 10 20 30 40 50 60 70 T -N ( m g · L -1) TU T-N 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 50 100 150 200 250 300 350 B O D ( m g · L -1) TU (S) BOD 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 20 40 60 80 100 120 140 160 C O D ( m g · L -1) TU (S) COD 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 10 20 30 40 50 60 70 T -N ( m g · L -1) TU (S) T-N 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 2 4 6 8 10 12 T -P ( m g · L -1) TU (S) T-P 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 2 4 6 8 10 12 T -P ( m g · L -1) TU (S) T-P 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 2 4 6 8 10 12 T -P ( m g · L -1) TU T-P 0 2 4 6 8 10 12

1 2 3 4 5 6 7 8 9 10 11 12 13

Site TU 0 2 4 6 8 10 12 T -P ( m g · L -1) TU (S) T-P summer

0 5 10 15 20 25 30 35 pH3 filtration pHi filtration Anion exchange pH11 filtration Cation exchange Mix exchange Spiking Mass balance

T o xi c U n it

Before Zn addition After Zn addition

0 5 10 15 20

Nov Dec Jan Feb Mar Apr May Jun Jul Aug Sep Oct

TU

Month

Output waste water (D)

After changing Fenton regent

Toxicity identification by Ulva test

Operational procedure

Space : <1m2

Person : 1 Time : 96 h

Facilities : temperature-controlled culture chamber, 24 well plates, pipets, tips, hole borer etc.

The Ulva kit

<Photo = jennifer R.S. Gordon, M.D>

 Physiological and biochemical damage

 Generation of reactive oxygen species (ROS)

(Rastogi et al., BBRC, 2010) DCFH-DA

395 nm 320 nm

Control

280 nm

• 3rd _{line of defense}

mechanisms • 2nd _{line of defense}

mechanisms • 1st _{line of defense}

mechanisms

 Production of quenching

 Antioxidant systems

 Migration  Mat formation

 Changes in morphology

to increase self-shading

 Production of

UV-absorbing compounds

• 4th _{line of defense}

mechanisms  Repair mechanism

PHOTOPROTECTIVE MECHANISMS UV

Seaweeds

Porphyra mats UV screening

compounds Photorepair Porphyra

UV-B

Porphyra yezoensis 0 0.1 0.2 0.3 0.4

280 300 320 340 360 380 400

- 133 species of domestic algae (17 of green algae, 34 of brown algae, 82 of red algae)

0 0.05 0.1 0.15 0.2 0.25

Palythine A-330 Shinorine P-334

Porphyra yezoensis (2001.1) Porphyra yezoensis (2001.3) Chondrus ocellatus (2001,7) Porphyra yezoensis (2001,2) Chondrus ocellatus (2000,1) Grateloupia filicina

Chondrus ocellatus (2001,5) Gloiopeltis furcata (2000,1) Pachymeniopsis elliptica (1999,12) Carpopeltis affinis (2001,2) Carpopeltis affinis(2000,1) Grateloupia turuturu Gymnogongrus flabelliformis (2000,1) Gelidium amansii (2000,1) Gloiopeltis furcata (2001,5) Schizymenia dubyi

Halymenia acuminata (2000,1) Pachymeniopsis elliptica (2001,1) Gigartina tenella Gelidium amansii (2000,1)

Lomentaria catenata(2000,1) Gymnogongrus flabelliformis (2000,1) Champia parbula Corallina spp.(2001,5)

Corallina officinalis Pelvetia wrightii

Pachymeniopsis spp.(2001,5) Lomentaria catenata(2000,1) Carpopeltis affinis (2001,5) Grateloupia filicina(2001,5) Grateloupia yezoensis(2001,5) Hypnea saidana

Corallina officinalis (2000,1) 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 Samples Conc ent ratio ns (%)

Porphyra yezoensis (2001.1) Porphyra yezoensis (2001.3) Chondrus ocellatus (2001,7) Porphyra yezoensis (2001,2) Chondrus ocellatus (2000,1) Grateloupia filicina

Chondrus ocellatus (2001,5) Gloiopeltis furcata (2000,1) Pachymeniopsis elliptica (1999,12) Carpopeltis affinis (2001,2) Carpopeltis affinis(2000,1) Grateloupia turuturu Gymnogongrus flabelliformis (2000,1) Gelidium amansii (2000,1) Gloiopeltis furcata (2001,5) Halymenia acuminata (2000,1) Pachymeniopsis elliptica (2001,1) Gigartina tenella

Gelidium amansii (2000,1) Lomentaria catenata(2000,1) Gymnogongrus flabelliformis (2000,1) Champia parbula

Corallina spp.(2001,5) Corallina officinalis

Pelvetia wrightii Pachymeniopsis spp.(2001,5) Lomentaria catenata(2000,1) Carpopeltis affinis (2001,5) Grateloupia filicina(2001,5) Grateloupia yezoensis(2001,5) Hypnea saidana Corallina officinalis (2000,1) Corallina officinalis (2000,1)

▷ Red algae (Rhodophyceae, Bangiales, Bangiaceae) ▷ Grows in the intertidal zone

▷ The most commonly eaten seaweed (ex. „Nori‟ in Japan, „Gim‟ in Korea) ▷ 60 to 70 species of Porphyra worldwide

 Porphyra

Porphyra yezoensis in vivo amplification

MAAs induction upon UV-A exposure

UV scatter

UV absor-bent

Absorb UV to inactive or intoxify

PABA (P-aminobenzoic 2 hydroxy-4 methoxurocanic acid, urocanic acid & benzophenone derivatives)

Severe side effects

Strictly regulated in most countries

Function

Type

Weakness

Reflect or scatter UV

Titanium oxide zirconia oxide zinc oxide

Little side effects

Less comfortable to apply on the skin

Function

Type

Weakness

Question 1

 Are Porphyra extracts strong UV absorbers?

 Structure and extinction coefficients of MAAs and synthetic filters

 Comparative analysis of MAAs in between Porphyra extracts and Helionori

UV–absorbing capacity

Porphyra extracts

Hellionori

 Comparison of UV absorbance between Porphyra extracts and ethylhexyl methoxycinamate

UV-absorbing characteristics

Question 2

UV-protective effects

Question 3

 Are there any other novel functions in Porphyra

 Melanin synthesis

 Tyrosinase inhibition assay

M el an in c on te nt o f c on tro l ( % )

Porphyra extracts (μL/mL)

T yr os in as e In hi bi tio n (% ) Whitening effects Arbutin (μL/mL)

+ α-MSH

- α-MSH

Re la ti ve A ct iv it y of M M Ps (% )

 Zymography for MMP2 & MMP9

Porphyra extracts (μL/mL)

 Collagen synthesis

Anti-aging effects

Porphyra extracts (μL/mL)

Product _(mg/mL) EC50 EC_(%) 50 CV

Porphyra

extracts 3.662 2.628

Tocopherol 8.879 0.663

 Comparison of ROS reducing power between Porphyra extracts and tocopherol

Antioxidant activity

Question 4

A bs orb an ce a t 5 40 nm 0 0.1 0.2 0.3 0.4 0.5

UVA - UVA + 100% hemolysis

UVA -UVA +

100% hemolysis

No photo- and cyto-toxicity of

Porphyra

Samples

3T3 NRU phototoxicity

Mean IC50(UV-)

(mg/mL)

Mean IC50(UV+)

(mg/mL)

PIF

IC50(UV-)/ IC50(UV+)

Result

Porphyra

extract > 1,000 > 1,000 1

Non- phototoxic

Samples IC50 Result

Porphyra

extracts >5,000μg/mL

Biochemical composition of

Porphyra

Component Contents Component Contents Component Contents

Moisture 11.1g Fiber 1.8g Vitamin B2 3.4mg

Protein 38.8g Potassium 2.1g Vitamin C 100mg

Non-fibrous 39.5g Calcium 390mg Vitamin E 4.3mg

Ash 6.9g Iron 12mg Niacin 9.8mg

Lipid 1.9g Vitamin B1 1.15mg Carotene 25mg

Question 5

 Is it economic and sustainable to obtain Porphyra extracts

Organisms [dry mass]

DDW/MeoH

/EtoH _{24-48h incubation}

40_C

Filtration

Evaporation Purification

Store at 40_C/-200_C

Application Photoprotection

Sun

Jangbong

Seochun

Shinan

Haenam

Jangbong Island

Relative amounts of

Porphyra

Re

la

tiv

e

am

ou

nt

of

P

or

ph

yr

a

ex

tr

ac

VS

 Floating-rafts method & Pole-type farming method

 Jangbong island

Years

V

o

lu

m

e

o

f

p

ro

d

u

cti

o

n

(M

T)

MAAs = 350-450 ton/yr

2012 2011

2010 2013 2015

Seaweed cosmetic brands

INcheon + COsmetic + BEAUTY; url : www.incobeauty.com connected to e-bay, Amazon, Yahoo, Rakuten, Taobao

Viral marketing

PR marketing

Mass media

PR marketing

Mass media

PR marketing

Arirang TV and famous drama

Radio

Social Media

PR marketing

Working with British company (Kilpatrick) for global marketing

Magazine (COSMOPOLITAN, PRIMA )

Export to China and Hong-Kong

Opening brand shops in China and

Export to the USA

Export to the UK

International licensing

China

Halal

New product developments

International cooperation

Market analysis

Cosmetic formulations PR marketing

Sale Profit

Malaga Univ. (Spain)

Incheon Nat‟l Univ. (Korea)

Ghent Univ. (Belgium)

SAG Gottingen Univ. (Germany)

There is hope from the sea,

but none from the grave

Taejun Han Prof., Director, CEO

Incheon National Univ., Institute of Green Environmental Research, Green Pioneer Co., Tel +82 32 850 5710, Mobile +82 10 3017 8248

hanalgae@incheon.ac.kr