Algae In Malaysia - Case Study.pptx

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Growing high value algae in a non algae business environment.

Malaysia - a case study. “T H E F U T U R E I S N O W” Frost & Sullivan 2011 Best Practice Award “Asia Pacific Green Excellence Award for Service Innovation in Algae Technology” BIONEXUS Status Award

ALGAE AS A BUSINESS Algae is already known to be an emerging and long-term sustainable source. This is due to its many benefits and business opportunities it offers: ALGAE GROW FAST ALGAE HAVE HIGH BIOFUEL YIELDS ALGAE CONSUME CO2 ALGAE DO NOT COMPETE WITH AGRICULTURE ALGAE CAN PURIFY WASTEWATERS

MICROALGAL BIOMASS CAN BE USED FOR FUEL, FEED & FOOD http://allaboutalgae.com/benefits/#sthash.NRk1qW4c.dpuf

BASIC ALGAE GROWTH CONDITIONS

Depending on the type of algae, these are the basic requirements for algae cultivation

MALAYSIA’S NATURAL CONDITIONS

• Temperatures fluctuate between 25 and 35 degrees during the year.

• Usually very hot and humid, especially in the major cities. • Between mid-October and the end of March the climate at the eastern side of the peninsula is affected by the rainy season or monsoon season

GROWING ALGAE IN MALAYSIA • Research on the production of microalgae biofuels is in the beginning stages and there is an urgent need for conducting more research on algae biofuels related to economic issues. • Geographically, the South China Sea borders Peninsular Malaysia in the east and both Sarawak and Sabah in the north that has natural advantages for algae culture. • Research and commercial applications of algae have gained more interest during the last few years in Malaysia. • Malaysia has a diverse knowledge based on algae, with more strength in biotechnology and bioengineering. • Industrial activity is also increasing slowly. Most of the active companies have valuable contributions and some existing collaborations with Malaysia universities; this inspires the interpretation of breakthroughs in academic fundamental research into industrial scale applications. Rajkumar R, Takriff MS (2015) Prospects of Algae and their Environmental Applications in Malaysia: A Case Study. J Bioremed Biodeg 7: 321. doi:10.4172/2155-6199.1000321

CASE STUDY: GROWING HAEMATOCCOCUS PLUVIALIS FOR THE PRODUCTION OF ASTAXANTHIN

WHAT IS ASTAXANTHIN?

Astaxanthin is a natural antioxidant with capability to scavenge free radicals Astaxanthin is a keto carotenoid, classified under Xanthophylls. It is a metabolite of Zeathanthin and / or Canthaxanthin. Astaxanthin is a colourful lipid soluble pigment Astaxanthin has two chiral centers/stereoisomers at the 3 and 3’ positions. There are three stereoisomers recorded so far in Astaxanthin 3R, 3’R, 2. 3R, 3’R and 3. 3S, 3’S

ASTAXANTHIN BENEFITS

Strong Antioxidant  Anti-Inflammatory  Provides Eye Fatigue Relief  Increase Muscle Endurance  Supports Healthy Blood Lipids  Aids in Wrinkle Reduction Astaxanthin is assimilated with lipoprotein and transported into the tissues.

Of several naturally occurring carotenoids, Astaxanthin is considered as one of the best carotenoids being able to protect cells lipids and membrane lipid proteins against oxidative damages (Olson, J.A., 2004. Pure Appl. Chem. 66: 1011 – 1016)

ASTAXANTHIN BENEFITS

• The colour of Astaxanthin is due to the extended chain of conjugated – alternating double and single bonds at the centre. The double bonds of hydroxyl and keto groups are responsible for antioxidant function of Astaxanthin. • Astaxanthin protects body tissues from oxidative and Ultra-violet damage through its suppression of NF-kB activation

(Lennikov, A., Nobyoshi, K. and Risa Fukase, 2012. Molecular vision 18: 455 – 464)

ASTAXANTHIN SOURCES

• The yeast, Xanthophyllomyces dendrorhous produces 3R, 3’R stereoisomers for the synthesis of Astaxanthin • Synthetic Astaxanthin obtained from petroleum products contains a mixture of all three stereoisomers • Haematococcus pluvialis Astaxanthin contains 3S, 3’S stereoisomers • Astaxanthin was approved by US, Food and Drug Administration (FDA) as a New Dietary ingredient in 1999

(Lennikov, A., Nobyoshi, K. and Risa Fukase, 2012. Molecular vision 18: 455 – 464)

HAEMATOCOCCUS PLUVIALIS Haematococcus pluvialis Haemato (Blood) Coccus (Seed) Pluvialis (Fresh Water)

Vegetative cell

Immature Aplanospore

Aplanospore

 Microalgae – includes Chlorella and Spirulina

 Found in standing pools and ponds world wide (non-GMO)

 Singlecell fresh water Microalgae (Plant source)

 During harsh environment conditions – Hibernates

WHY HAEMATOCOCCUS PLUVIALIS FOR ASTAXANTHIN?

 The freshwater unicellular microalga Haematococcus pluvialis synthesizes and accumulates the highest level of Astaxanthin compared to the other Astaxanthin producer organisms.  Commercially grown Haematococcus pluvialis can accumulate more than 40 g of Astaxanthin per kg of dry biomass.  Studies showed Astaxanthin’s health-beneficial effects in humans were performed on the stereoisomer found in Haematococcus, 3S-3’S.

 In Nutraceutical applications, scientists have proven that one of the main advantages of natural Astaxanthin esters is that the esterified form is inherently more stable than the free form, providing for a significantly longer shelf life without being oxidized.

WHY HAEMATOCOCCUS PLUVIALIS FOR ASTAXANTHIN?

The advanced commercial production of natural Astaxanthin from the microalga Haematococcus pluvialis supplies the market with high-quality products rich in Astaxanthin, suitable for human applications. Free Astaxanthin from H. pluvialis has 4.4 fold higher free radical scavenging activity (IC50 value of 8.1 microgram/ml) when compared to that of Astaxanthin esters (Kamath, 2007). Natural sources of Astaxanthin

FUNCTIONAL PROPERTIES OF ASTAXANTHIN AND OTHER COMPOUNDS

• Astaxanthin has shown both lipophilic & hydrophilic properties. • Astaxanthin donates the electrons and reacting free radicals to convert them to be more stable products & terminate free radical chain reaction. • Astaxanthin has a unique structure which enables to stay both in & outside cell membrane. • Astaxanthin gives better protection than beta carotene and vitamin C (Yamashita, 2013. Funct. Foods Health Dis. 3: 254 – 258)

THE RED COLOR FOUND IN NATURE IS ASTAXANTHIN

HEALTH BENEFITS OF ASTAXANTHIN

• For humans, Astaxanthin is a powerful antioxidant with broad health implications and unlike other antioxidants such as beta carotene, zeaxanthin, vitamin E, C, D. and selenium, Astaxanthin never becomes pro-oxidant in the body (Pearson, P., et al., 2006. BioDrugs 20: 271 – 273)

• Haematococcus pluvialis extract is known to inhibit the growth of human colon cancer cells by arresting cell cycle progression and promoting apoptosis (Palozza, P. et al., 2009. Cancer Lett. 283: 108 – 117) • Astaxanthin has shown excellent results for helping with skin moisture levels, smoothness and elasticity and preventing fine wrinkles and spots. (Camera, E., et. al., 2009. Exp. Dermotol. 18: 222 – 231)

HEALTH BENEFITS OF ASTAXANTHIN

• Bioavailability of Astaxanthin in human plasma was confirmed with single dosage of 100 mg (Osterlie, M., Bjerkeng, B. and Liaaen-Jensen, 2000. J. Nutr. Biochem. 11: 482 – 492) • Supplementation of Astaxanthin in human for eight weeks resulted in increased blood levels and improved the activity of killer cells which targeted to destroy the cells infected with viruses (Park, J.S. et al., 2010. Nutr. Metab. 7: 1 – 10)

• A clinical trial made on humans revealed that Astaxanthin helps diabetic retinopathy, macular degeneration, eye strain and fatigue and seeing in fine details (Sun, Z., et al., 2011. Food Funct. 2: 251 – 218)

HEALTH BENEFITS OF ASTAXANTHIN Oxygen Radical Absorbance Capacity (ORAC) values for H. pluvialis made Astaxanthin is about 3 times more than that of the synthetic product and about a third more than the yeast- made version (Nguyen, 2013). Table: Sustainability comparison on economical, environmental and societal impact of Astaxanthin production via chemical synthesis, yeast fermentation (Phaffia rhodozyma) and algal induction (H. pluvialis):

Per kg of Astaxanthin

Economical

Environmental

Production method

Raw material cost ($)

Land usage (Sq.km)

Energy cost ($)

Waste water

Emissions (Air)

Cost ($)

Oxygen Radical Absorbance Capacity (ORAC)

Human consum ption?

Chemical synthesis

40

10

26

0

2

2,000

33%

NO

Yeast fermentation

140

20

160

9

5

2,500

66%

Most

H. pluvialis synthesis

164

25

120

2

9

>7,000

100%

YES

Note: Green indicates the best option, yellow medium and red is the worse.

Societal

Comparisons of Astaxanthin content from Natural sources : Organism

Content % W/W dry wt

Reference

Green algae Haematocus pluvialis

2.0-3.0

Lorenz and Cysewski, 2000

Neochloris wimmeri

0.6

Orsa et al., 2000

Chloroccum

<0.2

Zhang et al., 1997

Nannochloropsis gaditana

<0.3

Lubian et al., 2000

Scenedesmus vacuolatus

0.01

Orsa et al., 2000

Chlorella zofingiensis

<0.01

Ip and Chen, 2005

Chlamydomonus nivalis

0.04

Bidigare et al., 1993

Phaffia rhodozyma

0.4

Jacobson et al., 2000

Yeast-Candida utilis

0.04

Miura et al., 1998

0.01

Yokoyama et al., 1995

Shrimp –Pandalus clarkii

0.015

Mayers and Bligh, 1981

Back snow crab-Chinoecetes opilio

0.011

Shahidi and Synowiecki,1991

Fungi

Bacteria Agrobacterium aurantiacum Animals

CULTIVATION SYSTEM OF H.pluvialis Astaxanthin production from H. pluvialis was developed on the industrial scale around the late 1990s. Many different systems have been developed for large scale production of H. pluvialis. First growth stage (Green stage): Usually facilitated by either closed indoor photo-bioreactors or enclosed outdoor systems. Closed system: -Environmentally controlled process with less biological and chemical contamination. -Much more popular and optimal growth conditions can be achieved. The cyst induction ‘reddening phase’ : Conducted via open raceway ponds, where nutrient deprivation and cellular stress are actually preferred. Industrial systems have utilized nutrient deprivation, high irradiance, and/or high temperature. Yields have hovered around 1-3% Astaxanthin harvested from cells.

Cell response to stress in the green alga Haematococcus Mode of action

Environmental Stresses:  Nutrient deprivation  Salt stress  High light  Low temperature  Drought  Aging

Slowdown of cell division

+

LIGHT

Excess of light Generation of ROS

Cellular sensing, mediators Activation of cell response Encystment

Motile cell

Palmelloid

1. Xanthophyll cycle 2. ROS quenching enzymes 3. antioxidants

Red cyst 21

Change in cell anabolism (lipids)

Astaxanthin accumulation

SYNCHRONIZED CULTURES AND LIFE CYCLE 40

10

1

0

Lag

enhanced productivity

10

10

20

30

40

50

60

70

1 0

80

20

40

60

Time (h)

Time (h)

Normal Light

High Light

22

80

100

CREATING A SYSTEM SUITABLE FOR HAEMATOCOCCUS PLUVIALIS

Continuous Cultivation •Green Stage to Red Stage is continuous

Acclimatization of Strain •Able to localize condition •Able to take up stress under tropical environment

Nutrients & Air •Preparation into system using non-conventional method of filtration •Multiple Filtration •No Contaminant •Humidity Control

Bio-Processing •Zirconia Beadmill •Controlled process to reduce oxidation of Astaxanthin from oxygen and light •Prevent Astaxanthin degradation to Astacene

Light •Lighting is optimized to compliment sunlight in tropical area •Improved growth during red stage with Sunlight & LED

ALGAETECH SYSTEM

Poly Bag •Prepared in-house under clean room system to avoid contamination •Multiple layers of plastic •Disposable •Specially Formulated

Media Preparation •Do not destroy nutrients •Improves Growth •Concentrated •Enzyme to induce stress for culture •Improve 30% Growth Density •Reduces deformation

CREATING AN ENVIRONMENT SUITABLE FOR HAEMATOCOCCUS PLUVIALIS

          

Fully Closed Cultivation Area Clean Room Facilities Temperature Control Light Intensity Control Centralised Aeration Media Distribution Facility Real-time Monitoring System 24 Hour Surveillance QA/QC Facilities Sufficient Power Supply 0.02 Micro Filtration System

CREATING A PROCESS FLOW SUITABLE FOR HAEMATOCOCCUS PLUVIALIS

UP-SCALING UP THE CULTIVATION PROCESS INOCULUM SCALE UP IN MOTHER CULTURE ROOM 1 Total flasks (10L) = 277 flasks Total blue bottles (17L) = 28 bottles Total PET bottles (5L) = 84 bottles Culture age:  0-7 days – 627L  8-15 days – 1,800L  16-23 days – 306L Parameters pH

Average data ±7.76-8.35

Cell Density

15,500-1,130,000 cell/mL

Temperature

20°C

Culture health

Juvenile lot, majority healthy some in dividing mode

UP-SCALING UP THE CULTIVATION PROCESS

CULTURE SCALE UP AND MATURATION IN GREEN STAGE ROOM Total bags = 1,092 bags Full capacity = 65,520L Current capacity = 58,080L (89% full) Culture age:  0-7 days – 26,340L  8-15 days – 0L  16-23 days – 35,880L  > 20 days – 35,880L Parameters

Average data

pH

±7.59

Sludge transfer

283L

Temperature

21.5°C

Culture health

Palmeloid high with contamination under control

UP-SCALING UP THE CULTIVATION PROCESS

CULTURE REDDENING IN RED STAGE ROOM Total bags = 2340 bags Full capacity = 140400L Current capacity = 129720L (92% full) Culture age:  0-7 days – 44520L  8-15 days – 50 400L  16-23 days – 28 440L  > 20 days – 6360L Parameters pH

Average data ±7.6

Light intensity

21,000 lux

Temperature

29°C

Culture health

Haema mostly in aplanospore stage with while some in brown color

WHY WE ARE BETTER?

Algaetech sees itself as competitive player due to its strength and ability in its innovative production process, without sacrificing high quality and standards. Controlled environment, higher productivity, shorter time PBR Bags offer maxiumum efficiency, and offer more sanitary solutions

• • • •

NO PLASTICISER NO HARMFUL TOXIN NO INSECT PARTS NO HEAVY METAL

Better control of gas transfer Lower evaporation More uniform temperature Less contamination Save space

CHALLENGES IN ASTAXANTHIN PRODUCTION Although natural sources have long been exploited for Astaxanthin production, it is still uncertain if natural Astaxanthin can be produced at lower cost than that of synthetic Astaxanthin or not. One of the major limitations with the H pluvialis production system is that the Astaxanthin gets trapped behind thick cells walls, thus complicating the extraction process and the production yields. Production capacity of H pluvialis is constrained by: • Its intrinsic slow growth, low cell yield • Ease of contamination by bacteria and protozoa, and • Susceptibility to adverse weather conditions.

H. pluvialis cannot be efficiently cultivated in dark heterotrophic mode and making the process economically less reasonable. (http://www.oilgae.com/non_fuel_products/astaxanthin.html#sthash.vyvgtBKR.dpuf )

ASTAXANTHIN PRODUCERS

• Algaetech will be the first company in Malaysia to commercialize Haema powder using the proven and latest technology for algae cultivation and bio process. • Currently there are 6 major pioneer prominent players in the market that produce Haema powder and Astaxanthin to meet worldwide demand. Company Cyanotech Corp. Mera Pharmaceuticals Inc. Algatechnologies LTD. Astareal (subsidiary of Bioreal, Inc.) Natural Astaxanthin Co. Ltd. (PA-Shun Group) Parry Neutraceuticals (Murugappa Group)

Location USA USA

Trademark BioAstin® Naturose® AstafactorTM

Israel Sweden

AstaPure® AstaCarox

China

NaturAstaTM

India

OleoResin 32

INDUSTRIAL PLAYERS

Currently there are a few major pioneer prominent players (production scale) in the market that produce Haema powder and Astaxanthin to meet worldwide demand.

Closed PBR + Raceway

Closed Tubular PBR

Cyanotech Hawaii Mera Pharma Inc

Algatechnologie s Ltd

Parry Nutra Ltd

Biogenic Co Ltd

Jingzhou Natural

Pigmentos SA Atacama Bio Natural Heliae Developm`ent LC

Closed Tanks/Cylinder PBR

Closed Plastic Bag PBR

Closed Flat Panel PBR

Fuji Chemicals Industry

Supreme Biotechnologies

Fraunhofer IGB @ Subitec

Algae Health

Maui tropical Algae Farm

Algae to Omega Inc

Algaetech International

BGG China Yunnan Alphy Biotech

MicroA AS

33

OUR ASTAXANTHIN FACILITY IN KUALA LUMPUR, MALAYSIA

Haematococcus Production Centre (1 acre)

Haematococcus Production Centre(5 acres)

2006 - Present - 1 Acre & Build up Area 25,000 sqft - R&D and Analytical Laboratory - 4 tons per year Haematococcus Cultivation - Pilot Bio-Processing Area - Mother Inoculum Room

2014 - Present - 5.013 Acres & Build up Area 150,000 sqft - World Class Laboratory Design - 38 tons per year Haematococcus Cultivation - High-Tech Bio-Processing Area 34 - Clean Room Mother Inoculum Area

TO BE OBTAINED

OUR ASTAXANTHIN FACILITY IN KUALA LUMPUR, MALAYSIA

A Brand New 150,000 sqft Production Facility

PREMIA ASTA-EX| OUR PRODUCTS 6000 times more powerful than Vitamin C!! 550 times more powerful than Green Tea!!

Why Astaxanthin? Astaxanthin is derived from a unique strain of microalgae called Haematococcus Pluvialis after acclimatizing to the local conditions in Malaysia. This type of microalgae strain is capable of producing high grade and 100% natural Astaxanthin. Unlike many antioxidants, which deal with one free radical at a time, our natural Astaxanthin handles multiple free radicals, with a unique molecular structure that lets it capture and neutralise multiple different free radical types.

Why Asta-EX?

RETAIL PRICE RM 98.00

The production of Astaxanthin at Algaetech is under strict control through use of advanced technologies to ensure high quality and potency of the product. The product is extracted into Oleoresin using Supercritical Fluid Extraction before being encapsulated into veggie caps. Launching of Asta-EX

• Product to be launched by Sept 2016 • To be sold in 300 pharmacies in across the nation by Sept 2016 • To be sold in retail chains and major pharmacies by Dec 2016

PREMIA ASTA-EXP| OUR PRODUCTS

Why Asta-EXP? The production of Astaxanthin at Algaetech is under strict control through use of advanced technologies to ensure high quality and potency of the product. The product is developed into powdered form using Cryogenic Vacuum Freeze Drying before being encapsulated..

ASTA-EXP PRODUCTS • • • •

Astaxanthin for Joints Astaxanthin for Cardio Astaxanthin for Sports (Active) Astaxanthin for General Health

COMING SOON! • • •

Launching of Asta-EXP Product to be launched by Sept 2016 To be sold in 300 pharmacies in across the nation by Sept 2016 To be sold in retail chains and major pharmacies by Dec 2016

ASTATUDE SKIN CARE| OUR PRODUCTS

Brightening Range 1. 2. 3. 4. 5. 6. 7.

ASTATUDE BRIGHTENING CLEANSING MILK ASTATUDE BRIGHTENING CLEASING FOAM ASTATUDE BRIGHTENING LOTION ASTATUDE BRIGHTENING SERUM ASTATUDE BRIGHTENING MOISTURIZER WITH SPF 35 ASTATUDE BRIGHTENING NIGHT MOISTURIZER ASTATUDE SPOT CORRECTOR

Anti Aging Range 1. 2. 3. 4. 5.

ASTATUDE CLAENSING MILK ASTATUDE CLEANSIN GEL ASTATUDE TONING LOTION ASTATUDE BOOSTER SERUM ASTATUDE DAY MOISTURIZER WITH SPF 35 6. ASTATUDE NIGHT MOISTURIZER

Specific Care Range 1. 2. 3.

ASTATUDE FACIAL SCRUB ASTATUDE FACIAL MASK ASTATUDE EYE SERUM

BUSINESS TO BUSINESS| OUR PRODUCTS

BUSINESS TO BUSINESS PRODUCTS • Haematococcus Powder – 2% - 5% • Astaxanthin Oloeresin – 5% & 10% • Astaxanthin Capsule – 4mg, 6mg & 8mg (Oleoresin) • Astaxanthin Capsule – 4mg (Powder) • White Label Astaxanthin • Booster Feed Additive for Poultry, Ruminants & Aquaculture • Other derivatives from microalgae

Launching of BUSINESS TO BUSINESS •

Effective immediately

THANK YOU