Biotechnology being multidisciplinary subject has applications in different areas. Marine Biotechnology is the field dealing with the uses of marine organisms for human use.

1. Module III

2. The Marine World
● More than 80% of living organisms on earth are found in the aquatic ecosystem.
The largest ecosystem on the planet is the ocean; it can be divided into photic,
pelagic, benthic, epipelagic, and aphotic zones.
● More than 40 000 different kinds of species are present in the marine environment,
and they are classified as microorganisms, seagrasses, algae, corals, and animals.
● The marine world is considered as a huge reservoir of various biological active
compounds. Marine organisms have the capacity to produce unique compounds due
to exposure to exceptionally different oceanic environments, such as temperature,
chlorophyll content, salinity, and water quality

3. Introduction
● Marine Biotechnology is carried out using biological resources which have come from
the marine environment rather than from the terrestrial environment.
● It is the application of scientific and engineering principles to the processing of materials
by marine biological agents to provide good and services.
● Marine biotechnology is an innovative field of research in science and technology
concerning the support of living organisms with marine products and tools.
● It is a novel way to produce genetically modified food, drugs, and energy to overcome
global demand.

5. Applications of Marine Biotechnology
1. Marine Aquaculture:
Fish is one of the most important
marine sources for protein
supplement in human food.
By applying marine
biotechnological tools, we may
be able to provide or improve
aquaculture procedures through
recombinant technology to
develop genetically modified
organisms , which could be
useful to overcome the global
food demand.

6. 2. Marine Natural products for medicine
Marine bioresources are huge reservoirs for various potential biological molecules, which have
tremendous potential as human medicines.Numerous marine compounds are isolated from
marine animals, algae, fungi, and bacteria with antibacterial, anticoagulant, antifungal,
antimalarial, antiprotozoal, antituberculosis, and antiviral activities.

7. 3. Marine Nutraceuticals
● Nutraceuticals are the products isolated from food sources which have a nutritional
value.
● Marine nutraceuticals can be derived from a vast array of sources, including marine
plants, microorganisms, and sponges. Marine nutraceutical products currently
promoted to various countries include fish oil, chitin, chitosan, marine
enzymes,chondroitin from shark cartilage, sea cucumbers,and mussels.
● For example, Fish protein such as collagens and their gelatin derivatives operate at
relatively low temperatures and can be used in heat sensitive processes such as
gelling and clarifying. Polysaccharides derived from alga, including alginate,
carrageenan, and agar types are widely used as thickeners and stabilizers in a
variety of food ingredients.
● Fucoidan is a complex-sulfated polysaccharide, which can be derived from brown
algae. It is important for its high bioactive properties, for example, antibacterial,
anticoagulant, antiviral, antitumor.

8. 4. Marine Biomaterials
● Marine bioactive substances for
healthcare would be the most
important and fastest-growing
sector.
● Non-toxic, biocompatible, natural
chitin and chitosan from crustaceans
have potential use in cosmetics,
food, and pharmaceuticals.
● Seaweeds are the abundant source
for polysaccharides, which are
commercial products (alginate, agar,
agarose, and carrageenan).

9. 5. Marine Bioenergy:
● Biofuels derived from marine algae are a potential source of sustainable energy that
can contribute to future global demands. The realization of this potential will require
manipulation of the fundamental biology of algal physiology to increase the
efficiency with which solar energy is ultimately converted into usable biomass.
● Anaerobic digestion of microalgae is a necessary step to make microalgae
biodiesel and biogas sustainable . The potential biomass sources for bioenergy are
photosynthetic microalgae and cyanobacteria. There are versatile marine organisms
that can be used in the production of biogas, biodiesel, bioethanol, and biohydrogen.

11. 6. Marine Bioremediation:
● Bioremediation is also an important area of marine environmental biotechnology.
Marine microorganisms have the capacity to degrade the variety of organic
pollutants.
● Pseudomonas chlororaphis produces pyoverdin, which catalyzes the degradation of
organotin compounds in seawater.
● Biopolymers and biosurfactants are also applied to environmental waste management
and treatment.

13. Marine Derived Pharmaceuticals

14. Introduction
● Marine-derived secondary metabolites have become a promising source
for the design and development of drugs.
● Marine derived-secondary metabolites, includes halogenated terpenes,
steroids and sterols, and polyphenols, have therapeutic potentials.
Marine proteins and lipids are also targeted to speculate on their role in
human health.

15. Marine Secondary Metabolites
● Exposure to extreme environmental conditions such as a broad range of variable
pHs, high hydrostatic pressure, and very low and very high temperatures may change
the morphology of marine organisms. So, the adaptation strategies of marine
organisms towards these conditions causes the production of secondary
metabolites.
● Marine natural products or secondary metabolites are quite species specific and have
been employed for the defense mechanisms against predation, infection, parasitism,
or interspecies communication and maintenance of homeostasis in marine
organisms.
● Most marine secondary metabolites play a physiological role in marine ecological
systems and are known to be involved in specific binding mechanisms with desired
target receptors.

16. Marine derived secondary metabolites
● Halogenated Terpene: Terpenes are derived from five carbon isoprene molecules
and described as a unique type of secondary metabolites
● Steroids and Sterols: A steroid comprises a characteristic arrangement of four cyclic
rings joined together. Sterols are a form of steroids and contain a hydroxyl attachment
at the 3-carbon position.
● Polyphenols: Polyphenols are a class of large secondary metabolites associated with
multiple phenol structural units. Polyphenols serve as a reactive species,tend to be
oxidized and are attributed as antioxidants, which can commonly be found in plant
materials.

17. Halogenated diterpenoid
Flexibilide
Poly hydroxylated steroid
glycosides
Phloro fucofuroeckol A and 8,8’-
bieckol
Halogenated Terpene Steroid Polyphenols

18. Halogenated Terpenes
Sr.
No.
Terpene Marine Organism Effect
1 Flexibilide and
Dihydroflexibilide
soft coral Sinularia flexibilis tissue necrosis
2 Halogenated terpenes Laurencia spp. Biofunctional effects
3 Brominated diterpene Malaysian laurencia Antibacterial activity
4 Palisadin
Palisol
Pacifigorgiol
Laurencia snackeyi Anti-inflammatory
effects

19. Steroids and Sterols
Sr.
No.
Steroid Marine Organism Effect
1 Isofucosterol and
clionasterol
marine sponge Petrosia
weinbergi
antiviral activities
2 Fucosterol Pelvitia siliquosa (brown
algae)
Antidiabetic and
antioxidant activity
3 Saringosterol Lessonia nigrescens (brown
algae)
inhibit the growth of
Mycobacterium
tuberculosis
4 Linckoside Linckia laevigata (Starfish) Neuritogenic and thus
might help to treat
damaged neuronal cells

20. Polyphenols
Sr.
No.
Polyphenol Marine Organism Effect
1 Eckol E.bicyclis (brown algae) Anti-diabetic properties
2 Eckol and dieckol Brown alga E.cava Inhibitory effect of
melanogenesis and
protective effects against
photo oxidative stress
induced by UV-B radiation
3 Dioxinodehydro eckol and
7-phloroeckol
Brown alga E.cava Anticancer drugs for
human breast cancers

21. Marine Proteins
● Most marine bioresources are composed of a higher content of proteins than other
components.
● Marine microalgae, muscle tissues of fish, shellfish, and other invertebrates show a
range of 15-65 w/w% proteins
● On the other hand, marine fish and shellfish waste as processing by products are also
considered to contain large quantities of high-quality proteins (10-23 w/w%).
● These sources are considered as being a potential source forming active proteins and
bioactive peptides
● They have high bioavailability and biospecificity to the targets
● Low toxicity, structural diversity, and little or no accumulations in the body
tissues

22. Marine Proteins

23. Marine Enzymes

24. ● One of the characteristic components in marine resources are lipids. The lipid
fractions of marine bioresources consist of polyunsaturated fatty acids (PUFA),
including omega-3 and omega-6 fatty acids.
● the consumption of seafood once or twice a week may have a protective effect against
coronary heart disease, may reduce the death risk by 36% and 17% from total
mortality due to a high content of docosahexaenoic acid (DHA) and
eicosapentaenoic acid (EPA) in the diet.
● These health benefits may bring about relievable effect against physiological
conditions such as diabetes, cancers, and inflammatory diseases.
Marine Lipids

25. Marine Lipids

26. Marine actinobacterial
metabolites & their
pharmacological
potential

27. ● Actinobacteria are either aerobes or anaerobes, motile or non-motile, and spore-/non-
spore forming, gram positive bacteria with a high G+C content.
● Marine actinobacteria are considered as a treasure house of secondary metabolites, and the
large numbers of these bioactive metabolites belong to the family Actinomycetaceae
including the genera of Streptomyces, Actinobaculum, and Acanobacterium and others are
commercially available due to their capability to produce novel bioactive molecules
● Actinobacteria are widely distributed in marine environments such as sponges, fish,
mollusks, mangroves, and seaweeds, besides seawater and sediments.
● Actinobacteria produce unique bioactive compounds like antibiotics; antioxidants; and
cytotoxic, antitumor, immunosuppressive, and cardiovascular agents with their unique
carbon skeletons that also provide a strong base for the synthesis of therapeutics
Introduction

28. Antimicrobial Activity of Actinobacteria
Sr.No. Compound Bacteria Effect
1 Bisanthraquinone
(Antibiotic)
Streptomyces sp. Inhibiting the growth of
methicillin resistant
S.aureus and vancomycin
resistant Enterococcus
faecalis.
2 bonactin the liquid culture of a
Streptomyces sp. BD21-2
antimicrobial activity
against both Gram-
positive and Gram-
negative bacteria as well
as antifungal activity

29. Anticancer Activity of Actinobacteria
Sr.No. Compound Bacteria Effect
1 Caprolactones
(Antibiotic)
Streptomyces sp. phytotoxicity and
promising activity against
cancer cells with
concomitant low general
cytotoxicity
2 salinosporamide A
(bicyclic beta-lactone
gamma-lactam)
Salinispora tropica orally active proteasome
inhibitor that induces
apoptosis in multiple
myeloma cells

30. Antitumor Activity of Actinobacteria
Sr.No. Compound Bacteria Effect
1 Marinomycins
(Antitumor Antibiotic)
Marinispora sp. antimicrobial activities
against drug resistant
bacterial pathogens and
demonstrate impressive
and selective cancer cell
cytotoxicities against
melanoma cell lines
2 Chinikomycins Streptomyces sp. antitumor activity against
different human cancer
cell lines,

31. Cytotoxic Activity of Actinobacteria
Sr.No. Compound Bacteria Effect
1 Mansouramycin C
(Isoquinoline quinones
antibiotic)
Streptomyces sp. Cytotoxicity for non-small
cell lung cancer, breast
cancer,melanoma, and
prostate cancer cells
2 Pyridinium
(salt antibiotic)
Amycolatopsis alba cytotoxic activity against
cancer cell lines of cervix
(HeLa), breast (MCF-7),
and brain (U87MG) in
vitro and also exhibited
antibacterial activity
against Gram-positive and
Gram-negative bacteria

32. Green Fluorescent
Protein (GFP)
● Characteristics
● Applications

33. Characteristics
● Green Fluroscent Protein is isolated from jellyfish Aequoria victoria
● GFP is a fluorescent protein that can be expressed in vivo. If GFP is exposed to light,
it emits a green fluorescent signal.
● GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a
minor one at 475 nm. Its emission peak is at 509 nm.
● MW of GFP- 269 KDa
● No. Of amino acids: 239
● GFP have a β-barrel structure. In the center of this β-barrel, there are 3 amino acids
(Threonine 65, Tyrosine 66, and Glycine 67). These acids and the cyclization and
oxidation of their backbone form a two-ring chromophore.

34. GFP Structure

35. ● GFP in cell biology and Biotechnology: It is used in various applications of
biotechnology such as protein fusions, imagining whole organism, and
transcriptional reporters.
● In the study of bacterial protein localization: Using GFP fluorophore, it is now
possible to monitor DNA and protein localization revealing the essential proteins
involved in sporulation and cell division and development that are dynamically
identified during the cell cycle. The GFP gene expression is utilized to examine the
primary cellular functions such as DNA replication, protein translation and
signal transduction.
● GFP as a reporter gene: GFP is considered as the most important and powerful tool
to monitor gene expression in different kinds of cells. It is favorably used as a
reporter gene since it does not require any supplement of an additional substrate.
Applications

36. ● GFP as active indicator
This protein is capable of acting as an indicator of the surrounding environment or in
the organelle. Phosphorylation sites of GFP are engineered to provide modifications in
fluorescent properties under a particular condition. The GFP engineered with
potassium channel is the first optical sensor to be genetically encoded in membrane
potential.
● GFP as fusion tag
The most thriving application of GFP is considering it as a genetically fusioned partner of
host proteins to observe their localization and destiny. The GFP gene is fused with the
gene encoding the protein of interest and expressed in the cell. The incorporation of GFP
can be done at either the amino or the carboxyl terminus of a protein. The fused protein
maintains its normal function along with acquired fluorescent property through GFP
expressing gene. Through GFP, all major cell organelles such as plasma membrane, Golgi
apparatus, nucleus, and endoplasmic reticulum are targeted.

37. Red Fluorescent
Protein (RFP)
● Characteristics
● Applications

38. ● The red fluorescent protein from Discosoma coral (DsRed or drFP583) holds great
promise for biotechnology and cell biology as a spectrally distinct companion or substitute
for the green fluorescent protein (GFP) from the Aequorea jellyfish.
● Once fully matured, the fluorescence emission spectrum of DsRed features a peak at 583
nanometers whereas the excitation spectrum has a major peak at 558 nanometers
● There were several problems associated with dsRed such as slow maturation,
fluorescence in green region, forming large protein aggregated in living
cells,inconsequential as a reporter.
● So, The second-generation RFP was developed by mutagenesis known as DsRed2,it
contains several mutations at the peptide amino terminus that prevent formation of
protein aggregates and reduce toxicity. In addition, the fluorophore maturation time is
reduced with these modifications.
Characteristics

39. ● Third Generation Ds Red (Ds Express): Further reductions in maturation time have
been realized with the third generation of DsRed mutants, which also display an
increased brightness level in terms of peak cellular fluorescence. Red fluorescence
emission from DsRed-Express can be observed within an hour after expression, as
compared to approximately six hours for DsRed2 and 11 hours for DsRed.
● A yeast-optimized variant, termed RedStar, has been developed that also has an
improved maturation rate and increased brightness.
● Several additional red fluorescent proteins showing a considerable amount of promise
have been isolated from the reef coral organisms. One of the first to be adapted for
mammalian applications is HcRed1, which was isolated from Heteractis crispa and
is now commercially available.It have an absorption maximum at 588 nanometers
and an emission maximum of 618 nanometers.

40. Applications
● Monitoring Physiological processes
● Visualizing protein localization
● Detecting transgenic expression in vivo

41. Chitosan: products and applications

42. Introduction
● Chitosan, sometimes known as deacetylated chitin, is a natural polycationic linear
polysaccharide derived from partial deacetylation of chitin.
● Chitin is the structural element in the exoskeleton of insects, crustaceans
(mainly shrimps and crabs) and cell walls of fungi, and the second most
abundant natural polysaccharide after cellulose.
● Chitosan is composed of β-(1-4)-linked D-glucosamine and N-acetyl-D-
glucosamine randomly distributed within the polymer.
● The cationic nature of chitosan is rather special, as the majority of polysaccharides
are usually either neutral or negatively charged in an acidic environment. This
property allows it to form electrostatic complexes or multilayer structures with
other negatively charged synthetic or natural polymers.

43. Advantages:
● Biocompatibility
● Non-toxicity
● Low allergenicity
● Biodegradability
Properties:
● Antitumor
● Antioxidant
● Antimicrobial

44. Chitosan Structure
Polymer of chitosan

45. Chitosan based drug delivery system

46. Application of chitosan and its derivatives in beverage industry

47. Application of chitosan and its derivatives in medicine and biomedicine

48. Application of chitosan and its oligosaccharides in dentistry

49. Application of chitosan and its oligosaccharides in veterinary medicine

50. Application of chitosan and its oligosaccharides in cosmetics

51. Application of chitosan and its oligosaccharides in agriculture

52. Application of chitosan in aquaculture

53. Application of chitosan and its derivatives in textile industry

54. Application of chitosan and its derivatives in pulp and paper industry

55. Application of chitosan and its derivatives in Biotechnology

56. Application of chitosan and its derivatives in chemistry

57. Application of chitosan and its derivatives in environmental chemistry

58. Application of chitosan in miscellaneous domain

59. Seaweeds for removal of metal pollutants

60. Introduction
● Biosorption is the process of adsorption of metal ions from wastewater.
● The biosorption processes occur when metal ions interact with the functional groups
present in biopolymers that are part of the biomass. Chemical groups such as amide,
hydroxyl, carboxylate, sulfonate, phosphate, and amino are responsible for the
quantitative adsorption of metals.
● Several interaction mechanisms such as complexation, coordination, chelation, ion
adsorption, cation exchange, and microprecipitation have been proposed as the
participants in the metal biosorption processes.
● One of the most promising types of biomasses suitable for their use as biosorbents is
marine algal biomass (seaweeds), which exhibit a high abundance in the oceans.

61. ● The biosorption capability of algae biomass is mostly related to their cell wall
chemical composition, which exhibits a fiber‐like structure and an amorphous
embedding matrix of polysaccharides such as alginates and fucoidan.
● In brown algae, alginates have a high affinity for divalent cations and sulfated
polysaccharides give account of the uptake of trivalent cations.
● The physical and chemical nature of the interaction between the metals and the
functional groups present in the biomass has been intensively studied, in order to
develop technologies for the sequestration of metals to clean, or to recover, valuable
metals from industrial effluents.

62. Seaweed Biomass
● Algae are autotrophic organisms that contain chlorophyll and carry out oxygenic
photosynthesis; they are widely distributed and have great diversity.
● Algae do not represent a formal taxonomic group of organisms, but a highly
heterogeneous collection of organisms of different evolutionary lineages and high
genetic diversity, which is reflected in the huge diversity that algae in morphological
terms, ultrastructure, ecological, biochemical, and physiological.
● Macroalgae, or seaweed, are a group of fast‐growing aquatic organisms including
about 9000 species. They are commonly classified into three groups according to the
color of the thallus, which correspond to the Chlorophyta (green algae), Rhodophyta
(red algae), and Heterokontophyta phylum, class Phaeophyceae (brown algae)

63. Algal divisions(seaweed Biomass) and its
characteristics

64. Process of removal of metal pollutants by seaweed biomass
Simplified scheme for industrial use of seaweed biomass. (1) Biosorption process; (2) demineralization
process; (3) secondary treatment such as pyrolysis, fermentation, HTL, gasification, among others

65. Mechanism of Biosorption
● Biosorption of heavy metal ions in wastewater using algae can be ecologically safer,
cheap, and efficient. Algae can be used for sorption of toxic and radioactive metal
ions and also to recover metal ions like gold and silver.
● The biosorption of heavy metal ions by seaweed biomass may occur by different
mechanisms such as ion exchange, complex formation, and electrostatic
interaction,being ion exchange the most important.
● Polysaccharides and proteins present in the algae cell walls provide the
metal‐binding sites . The sorption capacity of a seaweed cell surface to a specific ion
depends on several factors such as the amount of functional groups in the algae
matrix, the coordination number of the metal ion to be sorbed, the accessibility of
binding groups for metal ions, the complex formation, affinity constants of the
metal with the functional group, and the chemical state of these sites.

66. ● Considering the heterogeneity of the cell wall composition in different seaweed
species, the capacity of metal biosorption by the algal strains will vary. For
instance, brown algae with alginate in their cell wall composition have a high
biosorption affinity for lead ions. Alginate polymers are the primary responsible
for heavy metal ions sorption in brown algae, and their capacity to bind the metal
directly depends on the number of binding sites on this polymer. In a second place,
fucoidans play a key role for heavy metal sequestration.
● The functional groups present in the brown and green algae cell wall matrixes, such
as carboxyl, hydroxyl, sulfate, phosphate, and amine groups, play a dominant
role in the metal binding.

67. Comparison of Cu(II)
biosorption maximum
capacities of different types of
brown algae

68. Barophilic organisms and its applications
● The deep-sea is regarded as an extreme environment with high hydrostatic
pressures (up to 110 MPa), predominantly low temperatures (1°C–2°C) but
with occasional regions of extremely high temperature (up to 375°C) at
hydrothermal vents, darkness, and low nutrient availability.
● Barophilic bacteria are defined as those displaying optimal growth at pressures
>40 MPa, whereas barotolerant bacteria display optimal growth at pressure
<40MPa and can grow well at atmospheric pressure. Barophiles are also known
as piezophiles.
● An example of a high-pressure habitat is the deep-sea environment, such as ocean
floors and deep lakes where the pressure can exceed 380 atm.

69. ● Halomonas salaria, a gram-negative proteobacteria, is an example of an obligate
barophile. It needs a pressure of 1000 atm.
● Many barophiles are sensitive to ultraviolet rays and are susceptible to UV radiation.
They lack the essential mechanisms of DNA repair to counter the effects of UV
radiation. Thus many of them grow in darkness.

70. Applications of Barophiles
● Enzymes produced by barophilic bacteria can function at high pressure and
may be useful for high pressure bioreactors, toxic clean-up in deep sea and
high pressure food processors.
● Barophiles can be used in microbially enhanced oil recovery process.
● Proteins from barophiles are used for the production and sterilization of food
items at varied pressure conditions.