Macro and micro algal natural products

1. Dr. T. Citarasu
Associate Professor
Tele-fax: + 91-4652-253078
Mobile: +91-9994273822
Email: citarasu@gmail.com
citarasu@msuniv.ac.in
Website: www.msuniv.ac.in
MANONMANIAM SUNDARANAR UNIVERSITY, TIRUNELVELI

2. BIO-PROSPECTING OF MARINE
ALGAL NATURAL PRODUCTS AND
THEIR HEALTH IMPORTANCE

3. Importance of Marine Biotechnology
Contribute to oxygen cycle & help to regulate the
Earths’ climate
Explore medicines, cosmetics, nutraceuticals,
enzymes, industrial products, bio fuels, foods,
aquaculture etc.
Defined as “ Application of scientific & engineering principles to the processing
of materials by marine biological agents to prove goods and services”.

4. 7516.6
km
News Courtesy :
Daily Thanthi 23.02.2014
Strengthen our marine biotechnology research…….
Indian Council of Agricultural Research (ICAR)
Department of Biotechnology (DBT)
Ministry of Earth Sciences (MOES)
Ministry of Environment and Forest (MOEFS)
Council of Scientific & Industrial Research (CSIR)

5. Need for Marine Bio-prospecting
Repeated search in terrestrials leading to known compounds
Marine ecosystems are unique and rich reservoir of biodiversity with an
enormous potential towards improving the quality of human life.
Marine organisms have highly developed defense system in order to survive
in the hostile conditions such as extreme temperatures, varied pressures
(low or high), low energy and lack of sunlight

6. Multi-OMICS methodologies: Genomics, Transcriptomics, Proteomics,
Metabolomics, Metagenomics and Meta transcriptomics
Production and analysis of massive biological data
Once, the compound of interest is identified, bioactivity screening,
heterologous expression, genetic engineering and/or chemical synthesis can
follow
Characterize the activity and produce the desired bioproducts with
pharmaceutical, nutraceutical, cosmeceutical and biofuel production
applications.

7. MARINE ALGAL BIO-PROSPECTING

8. ALGAE IN THE WORLD
Sea, rivers and lakes, on soil and walls, in animal and plants
Symbiosis with plants, animals, bacteria and Marine organisms
Macro algae (seaweeds) occupy the littoral zone, which included green,
brown and red algae
Micro algae are found in both benthic and littoral habitats and also
throughout the ocean waters as called phytoplankton

9. MICRO ALGAE
Microscopic photosynthetic organism called phytoplankton found in fresh
and marine waters , plays a key role in food chain
Similar photosynthetic mechanism as like plants & they utilize sunlight,
atmospheric CO2 and nutrients in the water to produce biomass
Classification based on morphology, pigmentation , chemical nature of
photosynthetic product & organization of photosynthetic membranes
Diatoms (bacillariophyta), dinoflagellates (dinophyta), green and yellow–
brown flagellates (chlorophyta; prasinophyta; prymnesiophyta, cryptophyta,
chrysophyta and rhaphidiophyta) and blue–green algae (cyanophyta).
Micro algae are found in both benthic and littoral habitats and also
throughout the ocean waters as called phytoplankton

10. MICRO ALGAL COMPOUNDS & ITS BENEFITS

11. PHARMACEUTICAL & COSMETIC APPLICATIONS
Carotenoids and Pigments
Powerful antioxidants, nutraceuticals use in Eye applications
Preventing Cancer , Immunostimulant & veterinary use (Aquaculture use) & cosmetics
Commercially available
Dunaliella salina

12. Astaxanthin
Isolated from Micro algae
Powerful antioxidants in aquaculture
Fucoxanthin
Promote DHA & omega-3 fatty acids
Beta carotenoid
Found in many algae
Colored carotenoids
Dunaliella salina
Phytoene
Phytofluene
Lutein
Zeaxanthin

13. ANTIBACTERIAL COMPOUNDS
C-lactone malyngolide
(Lyngbya majuscula)
ANTIFUNGAL COMPOUNDS
Majuscuiamide (cyclic depsipeptide)
Gambieric acid
(Gambierdiscus toxicus –Dinoflagellate)
IMMUNOSUPPRESSIVE COMPOUNDS
Lipoproteins, microcolins A and B
ANTICANCER COMPOUNDS
Curacin A – anticancer (antimitotic agent)
Cytotoxic pentapeptides:
majusculamide D
Deoxymajusuculamide D
Amphidinolide B
(dinoflagellate, Amphidinum sp.)
ANTIMICROBIAL & ANTICANCER COMPOUNDS

14. FATTY ACIDS AND HIGH VALUE OILS
Very Long chain PUFA (vlc PUFA)
Eicosapentaenoic (EPA)
Docosahexaenoic acid (DHA)
Arachidonic acid (AA)
Flexibility, fluidity and selective permeability
to cellular membranes
Brian development
Beneficial for the cardiovascular system
Nutraceutical in human and animal health

16. Antibiotics (fatty acids, bromophenols, tannins, terpenoids, polysaccharides,
alcohols)
Phycobiliproteins (Phycocyanin & phycoerythrin)–Natural dye for cosmetics
(lipsticks & eyeliners)
Algicides –toxin (neurotoxic and hepatotoxic)
Vitamin and vitamin precursors (ascorbic acid, riboflavin and tocopherol)
Chlorella and Spirulina are used in skin care, sun protection and hair care
products

17. ALGAL SMALL MOLECULES
Group of secondary metabolites synthesized by biosynthetic gene clusters
(Enzyme systems producing products such as antibiotics)
Cyclic peptide (microcystin), depsipeptide ( majusculamide)
Lyngbyatoxin-A ( Lyngbya majuscule) – antifungal activity
Cryptophycin from Nostoc sp. shows a fungicidal activity &microtubule
depolymerizing agent for Cancer cells.
Heterocyclic compounds, polycyclic ethers, oxygenated polyketides and
macrolides from dinoflagellates
Dinoflagellate toxins like saxitoxin , neosaxitoxin and gonyautoxins are sodium
channel blockers

18. ALGAL BIOSURFACTANTS
Extracellular Amphiphilic compounds
Glycolipids, lipopeptides, fatty acids, polymeric surfactants & particulate
natures
Broad range of applications
Antibacterial, antiviral, antifungal, anti-adhesive, anti-cancer, antioxidants,
adjuvant etc
Arthrospira sp., Synechococcus nidulans, Chlorella minutissima, C. vulgaris and
C. homosphaera used for various industrial and pharmaceutical applications

19. ALGAL BIO POLYMERS AND BIOPLASTICS
Typically, long chain polymers, present in the algae lipids are used for making
bioplastics
Algae is attractive for bioplastics because of its high growth rate and high protein
content.
The plastics market is worth more than $400 billion and has grown at an average of
3.5% per year over the last two decades.
Naturally occurring polymer have many applications in medical, pharmaceutical and
food industries
Biodegradable Poly hydroxy alkanoates (PHA) are promising biopolymer which can be
alternative to petroleum based polymer
PHB poly-(hydroxy butyrate) is the most abundant PHA which is homopolymer of
hydroxybutyrate that presents in various cyanobacteria such as Chlorogloea fritschii,
Spirulina sp., Aphanothece sp. & Gloeothece sp.

20. MICRO ALGAE AS FOOD
Human Food – Good Nutraceuticals
High protein content
Chlorella and Spirulina, Dunaliella, Haematococcus, Schizochytrium, Scenedesmus,
Aphanizomenon, Odontella, and Porphyridium are health-food
Chlorella, the prevention and treatment of heart diseases with the Spirulina, Vitamin A
precursor (Dunaliella), anti-inflammatory (Haematococcus), treatment of brain and
heart disorders (Schizochytrium),
Dunaliella bardawil could improve the drug’s effect on HDL-cholesterol levels in
patients
30 % of the food in Astronauts is algae due to high protein

21. Microalgae as animal feed
Traditional uses of microalgae in Aquaculture
Food for
All growth stages of bivalves
Crustacean
some larval stages
Cattles
Some fish larvae
Zooplankton
used as food for
larval crustaceans
and fish
Fatty acid enrichment of zooplankton
Dunaliella sp.
Chlorella sp.
Nanochloropsis sp.

22. MICRO ALGAE IN SPACE RESEARCH
Algae produce oxygen in space
Chlorella and Euglena grown in
photobioreactor is used to produce oxygen
in the space for the Astronauts using their
waste and help them for long term
expedition in space
NASA and many other space research
station in the world open opportunities for
planktologist

23. ENERGY PRODUCTION
Bio-hydrogen production
Photosynthetic microorganisms are attracting
nowadays by high photosynthetic conversion
efficiencies diverse metabolic capabilities, superior
growth rates, and ability to store or secrete energy-
rich hydrocarbons.
Two methods; (1) microalgae use the light energy
to produce hydrogen from water, (2) bacteria capable
of fermenting the carbohydrates (either structural or
stored in the microalgal cell wall) are utilized
to produce hydrogen.
Manipulation of competing metabolic pathways by
modulating the certain key enzymes such as
hydrogenase and nitrogenase may enhance the
evolution of H2 from photoautotrophic cells
Several photobioreactors have been developed for
large-scale biomass and hydrogen production

24. Algal Methane Production
In a biogas reactor the substrates used are converted in several reaction
steps into biogas, consisting mainly of the components CH4 and CO2
The biogas yield of plants is generally limited by the greater or lesser
proportion of lignocellulose, which is difficult to recycle.
However, the use of microalgae with a low lignocellulose content, for
example Chlorella vulgaris, Phaeodactylum tricornutum and Spirulina
platensis, permits an almost complete utilization of the organic substance.
.

25. ALGAL OIL PRODUCTION
Storing the Sun’s Energy
(Photosynthesis)
What is needed Storage of Energy
Sunlight Lipids and oils
CO2 Carbohydrates
Nutrients

26. How to produce more oils ?
The lipid content of an algae species can be increased by imposing a stress
factor, such as starving of nitrogen or silicon.
In this manner, lipid fractions up to 70-85% have been reported for nitrogen
starvation and 60% for silicon starvation

27. Algal Cell Harvest
Centrifugation- small microalgae
Filtration- different size of algae
Flocculation- Change of pH
Acid flocculation – using alum and sulphuric
acid
Alkali flocculation – using NaOH and KOH
Autoflocculation – Interrupting the CO2 level

28. Algal Oil Extraction
Mechanical methods: Expeller press, with vegetable oils, with chemical
solvents
Osmotic methods: Sudden reduction in osmotic pressure
Ultrasonic assisted extraction: ultrasonic reactor, ultrasonic waves crate
the rupture of cell walls
Sonoenzymatic treatment: Accelerates extraction and increases yields
Chemical methods: Algal powders + organic solvents (1:2)
Supercritical fluid extraction: liquid CO2 With different pressure
Enzymatic extraction: Enzymes to degrade the cell walls

29. Transesterification
A reaction between an ester of one
alcohol and a second alcohol to
form an ester of the second alcohol
and an alcohol from the original
ester, as that of methyl acetate and
ethyl alcohol to form ethyl acetate
and methyl alcohol
Methods for produce biodiesel

30. Advantages
Rapid and faster growth rates than terrestrial crops
The per unit area yield of oil from algae is estimated to be from between
5,000 to 20,000 gallons (18,927 to 75,708 litres) per acre, per year; this is 7 to
31 times greater than the next best crop, palm oil (635 gallons or 2,404 litres)
Certain species of algae can be harvested daily
Algal biofuel contains no sulphur, non-toxic and highly bio-degradable

31. Health aspect of algal oils
The oil extracted from
Crypthecodinium cohnii contains 40-
50% DHA useful for infant formula
Schizochytrium produce algal oil
known as DHA Gold for use as an
adult dietary supplement in food and
beverages as well as animal feeds
(OmegaTech (USA)).
Other applications include food for
pregnant and nursing women and for
cardiovascular health

34. Effluent Remediation
Phycoremediation
Effectively remove ammonia (75 %) ; Nitrite (84 %) ;
Phosphorus (89)
Removal of nutrients
Remove excess nutients and CO2 in waste water
systems
Solve BOD problems
Excess algal biomass may harvested
Removal of organic pollutants
Chlorella, ankistrodesmus in paper mills & olive oil
mills
Algae can produce O2 to bacteria and bacteria
degrade the nutrients
Diatomite- silicon compound for chelating
PHYCOREMEDIATION

35. ALGAL ASSOCIATED MICROBES AND ITS IMPORTANCE
The symbiotic relationship helps to enhancing the efficiency of micro
algal biomass and production of valuable organic compounds
Enhancement in the intracellular levels of carbohydrates, lipids and
pigments in microalgae (Emiliania huxleyi & Scrippsiella trochoidea)
Interactions can be beneficial to the massive production of microalgae
and algal products.

37. ALGAL GENOMICS
Genome sequences can be
instrumental in the reconstruction of
metabolic pathways
The metabolic pathways help to study
the production of various secondary
metabolites and increased production
etc
Sequence analyses provide information
on the evolution of different groups of
algae
Potential to develop new and
economically important products and
technologies.

39. Transcriptomics
Transcriptomes are sequenced among the available micro algal genomes
Transcriptome sequences have been deposited in public databases GenBank
A specific web-based database, named AlgaePath, is available for some of the
algae (http://algaepath.itps.ncku.edu.tw/)
AlgaePath integrates gene information, biological pathways and NGS datasets
Considering the great market interest of MNPs, the transcriptomic approach has
been used in order to reveal the gene pathways responsible for their synthesis.

40. Proteomics
Genomics and transcriptomics alone are insufficient to understand the complex
biology of microalgae, and should be complemented with a proteomic approach
Proteomics explores the mechanisms involved in many biological processes and
network functions by providing information on proteins, including post-
translational modification, sub cellular localization and protein-protein
interaction
Proteomics analysis helps to biofuel, lipid bodies, cytoplasmic lipid droplets, β
carotene-rich plastoglobuli , biosynthesis and regulation , nitrogen depletion,
membrane protein etc (http://alga-pras.riken.jp/)

41. Metabolomics
Metabolites are dynamic, and their properties and levels of biosynthesis depend
on genetic and/or environmental changes
Their synthesis can be triggered by physiological and/or environmental stimuli
(e.g., the presence of predators, nutrient depletion or starvation and light etc
For example metabolomic and lipidomic profiles of Nannochloropsis salina
under cold stress, and observed a total fatty acids accumulation
Metabolomics information available in Kyoto Encyclopedia of Genes and
Genomes (KEGG), PubChem and MetaCyc etc.

42. TRANSGENIC ALGAE
Possibility of manipulating genes is an attractive opportunity, and could lead to
leaps forward in reducing costs and improving production efficiency
Recombinant DNA technology and recently the gene editing approach, CRISPR
etc
The engineered microalgae can be modified to enhance growth properties,
photosynthetic efficiency or enhanced production of metabolites
Recombinant products including vaccines, oils, novel carotenoids, antibodies,
insecticidal proteins and biohydrogen etc

43. Nuclear transformations in
chlorophytes, diatoms and
dinoflagellates
Agitation with glass beads,
electroporation, microparticle
bombardment and
Agrobacterium mediated
transformation are used to
transfer foreign genes into
unicellular eukaryotic
microalgae
Chloroplast transformation is
achieved in Euglena gracilis
and unicellular red alga,
Porphyridium sp. for the
production of biosynthetic
enzymes

44. MACRO ALGAE (SEAWEEDS)
Seaweeds belong to the lower plants, do not have roots, stems and leaves.
Instead they are composed of a thallus (leaf-like) and sometimes a stem
and a foot and some species have gas-filled structures to provide buoyancy.
They are subdivided in three groups, the red (Rhodophyta), green
(Chlorophyta) and brown (Phaeophyceae) seaweeds
Top cultivators: China, France, UK, Japan, Chile, North Korea, Philippines,
Norway and Indonesia etc.

45. HIGH VALUE ADDED PRODUCTS
Bio molecules
Iodine
Algin
Mannitol
Glycerol & Proline
Polymers
Phyto colloids (Alginate, agar, carragennan etc)
Cyanobacterial origin –Cyanophycin
Hydrocolloids
Thickening, Emulsify & stabilizers
Also a polymer D-mannuronic & l-guluronic acid
Laminaria, Ascophyllum & Macrrocystis

46. Alginate
Polyuronic saccharaide, inhibit Virus
Laminaria and Fucus
D- mannuronic and L-galuronic acid
Emulsifier in food industries
Sargassum & Gracilaria sp
Fresh seaweed washed & Dried
Pulverize to get powder
Treat with 1N HCl (pH 2-3)-ON
Wash with water & add Na2Co3
Filter & bleach with 2.5 % NaClo
Evaporate @ 60° C
Pure sodium alginate

47. Carrageenan
Isolated from sea weeds
Sulfated polysaccharides
Powerful antioxidants
Broad range of applications in food
and cosmetics

48. Fucoidon (Foo–Koy–Den)
Sulfonated polysaccharides
Antioxidants & immunostimulants
Brown sea weeds
Blood purifier
Reduced Cholesterol
Fight off viral infections
Scavenge free radicals
Liver tonic
Anticancer

49. Ulvan
Composed of rhamnose, glucuronic acid,
iduronic acid, xylose and sulfate
Green seaweed, Ulva and Enteromorpha
Good gelling agent
Prevent biofilm formation
Good Nutraceuticals
Antioxidants
Anti-thrombotic activities
Treatment of gastric ulcers
Anticancer
Immunostimulants

50. Laminarian
Water-soluble polysaccharide
(1,3)-β-D-glucan with β (1,6) branching
and contains 20–25 glucose units
Regulator of intestinal metabolism
Tumor-inhibiting agent
Antibacterial agent
Protection against severe irradiation
Boosting the immune system
Reducing cholesterol levels in serum

51. Cadalmin Green Algal extract
Cadalmin Green Algal extract contains a unique
blend of 100% natural marine bioactive anti-
inflammatory ingredients extracted from selected
seaweeds or marine macro algae with a patented
ecofriendly “green” technology .
The product is effective to combat arthritic pain and
inflammatory diseases in human beings
Patients suffering with chronic joint pain and
arthritis clinical trial-based studies revealed that
more than 98% of the respondents were satisfied
with the product with about 70-85% relief in joint
pain and arthritis

52. PHARMACEUTICAL & COSMETIC APPLICATIONS
Bioactivity Seaweeds Compounds
Antioxidant Cymopolia barbata Cymopol
Avrainvillea longicaulis Avrainvilleol
Halimeda opuntia Polyphenols
Undaria pinnatifida Fucoxanthin
Anti-inflammatory Laureafincia claviformis Pacifenol
Stypopodium flabelliforme Styfotriol
Undaria pinnatifida Fucoidon
Anticancer Gracilaria corticata Polysaccharides
Gracilaria tenuistipitata
Plocamium telfairiae
Capsosiphon fulvescens
Sargassum oligocystum

54. FOOD APPLICATIONS
Human Consumption
Seaweeds are utilized as food in China, Japan, Korea, Philippines,
and several other Asian countries as human food
They provide a large profile of natural vitamins, minerals, and
essential fatty acids for human health
Porphyra, Monostroma sp., Ulva sp., Laminaria sp., Undaria sp,
Hizikia fusiformis, Chondrus crispus, Caulerpa sp., Alaria
esculenta, Palmaria palmata, Callophyllis variegata, Gracilaria sp.
and Cladosiphon okamuranus

55. Sea Lettuces
Genus Ulva, a group of edible green algae that is widely distributed along the
coasts of the world's oceans.
Food source for humans in Scandinavia, Great Britain, Ireland, China,
and Japan
Used as salads and cooked in soups.
High in protein, soluble dietary fiber, vitamins and minerals, especially iron.

56. Ulva reticulata

57. Animal Consumption
Seaweeds are grazed by sheep, horses, Pig, cattle, and a nutritive
feed for fish and shrimps
Also a good food for Cats, dogs, aquarium fish, ornamental birds,
poultry and breeding bulls
Ulva sp., Porphyra sp., Palmaria palmata, Gracilaria sp., and Alaria
esculenta are used as animal feed
Hypnea cervicornis and Cryptonemia crenulata rich in protein used
for shrimp diets

58. FERTILIZER
Broad spectrum fertilizer that is rich
in beneficial trace minerals and hormones that
stimulate plant growth
High carbohydrates which are essential
building blocks in growing plants, and low in
cellulose so it breaks down readily
Trace elements found in organic seaweed
fertilizers include magnesium, potassium, zinc,
iron and nitrogen—all of which are beneficial to
plants
Ecklonia
Ascophyllum – Agri & Horticulture purposes

59. Bioremediation
Removal of heavy metals
Sargassum sp special affinity to heavy metals
S. vulgare, S. fluitans and S. filipendula , S. crassifolium
Effectively uptake Cd & Cu etc
Binding capacity due to alginic acid, with carboxyl groups and sulfated
polysaccharides, as well as sulfonic acid
Seaweed generates O2 which benefits hypoxic dead zones. Nutrients such as
ammonia, ammonium nitrate, nitrite, phosphate, iron, copper, as well as
CO2 are rapidly consumed by growing seaweed.

60. Algal Identification
Zooplankton culture
Algal associated microbes
Carotenoid production
Algal oil production
Algal secondary metabolites
Aquaculture live feeds
Other compounds
Research Training @ CMST

61. Aquaculture companies
Amalgam Enterprises
Arbee Agencies
Arianee Landscapes
Avanti Feeds Ltd
Bright and Associates
Castlerock Group Of Companies
CIBA
Chirag International
Cochin Frozen Food Exports Ltd
Geo Seafood
Guybro Chemicals
L G Sea Foods
Carotenoid Companies
Allied Biotech Corporation
Carotenoid Technologies
Sociedad Anonima
Novepha Company
Agro Bio Chem (India) Pvt Ltd
Yunnan Green A Boiolgical Company
Algart International Inc.
Proalgen Biotech Ltd, Chennai
MNC Globle
Divis Laboratories Ltd
Agro BioChem (India) Pvt Ltd
BOBO Corp
Hovid Bhd
Global Opportunities

62. Job Opportunities in Algal Industry
Shrimp farm and Hatcheries
Algal oil industries
Carotenoids industries
DHA Production industries
Space research
Health product industries
Cosmetic industries
Biofertilizer industries

63. Algal oil producing companies
Aquaflow Binomic Corporation (ABC)
Aquatic energy
Circle Biodiesel and Ethanol Corporation
Amalgam Enterprises
Green Star Products Inc., (GSPI)
Inventure Chemicals
LiveFuels OriginOil, Inc.,
PetroAlgae
Solazyme, Inc.,
Solix Biofuels
Texas Clean Fuels

64. CONCLUSIONS & FUTURE DIRECTIONS

65. Marine Algal derived compounds provide a novel and rich source of
chemical diversity that can contribute to design and development of new
and potentially useful pharmaceutical agents.
Algal genomics provided good opportunities to identify and characterize
high-value bioactive compounds derived from marine microalgae.
Utilizing our salt lakes and coastal area , the production of macro and
micro algal may increased. This approach help to more income generation
to our country.
Consumption of Seaweed foods helps to improve our health status and
also act as good nutraceuticals for many diseases.
A proactive interaction between researchers, the pharmaceutical marine
biotech sector and government regulating agencies is crucial to the
incorporation of this challenging new tool in clinical medicine.

66. THANK YOU