VIETNAM NATIONAL UNIVERSITY INTERNATIONAL UNIVERSITY

MARINE ASSIGNMENT
Marine Biotechnology Applications: From Laboratory to Daily Life

Submitted to Dr. Gay Marsden in Partial Fulfillment of the Requirements for Marine Biology course Second Semester, 2010 - 2011

Student Name: Nguyen Bao Ngoc Student ID: BTIU08163 Date: May 27, 2011

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Pharmaceuticals: Adenine arabinoside (AraA) or spongoadenosin is the first antiviral compound derived from marine creature, which is active against Herpes viruses via the inhibition of viral DNA synthesis. In fact, AraA is an analogue of the spongosine compound which was firsly derived from marine sponge (Cryptotethya crypta) in 1949 by W. Bergmann. Interestingly, it was found that there was a novel base, methoxyadenine combined with ribose in the nucleoside instead of the classical adenine. Due to its interesting structure and the widespread interest in nucleosides at that period, the compound was widely studied. As a result, the antiviral activity of the nucleosides was soon demonstrated in 1964 by Privat de Garilhe and De Rudder and was further confirmed its clinical usefulness in 1976 by Whitley.This led to the synthesis of AraA, which had been approved by FDA and commercialized under the name of Vidarabine. AraA was chemically synthesized analogous to spongosine so that it is improved in the chemotherapeutic properties, therefore, had higher activity against Herpes viruses. One promising method had been reported to produce AraA in large scale for industrial production was chemico-enzymatic method (Utagawa et al. 1980) in which the microbial enzyme acted as a bio-catalyst during the chemical synthesis of AraA. The system included two steps: uracil arabinoside (AraU) was chemically synthesized in the first steps and in the second step, AraU and adenine were enzymatically conversed to form AraA.

Fig. Cryptotethya crypta (left) and structures of spongosine extracted from sponge and AraA formulated for antiviral treatment (right)

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Unfortunately, the marketing of AraA drug now has been stopped due to its less efficiency and more toxicity compared to the new discovered drug, acyclovir (Zovirax). Nutraceuticals: Green Lipped Mussel is a natural anti-inflammatory supplement, which contains marine lipids and is extracted from Perna Canaliculus, a green-lipped mussel native to New Zealand. For centeries, based on observations of Maori tribes in New Zealand, GLM was known as able to help maintain good health since it was rich of vitamins, minerals and extremely valuable Omega3 fatty acids. Not until after 1970s, green-lipped mussel powders were firstly approved to be commercially available products for alternatively treating arthritis. However, it was shown that the efficacy of these powders could be influenced when being kept overtime due to the oxidation of their non-polar lipids. Therefore, fifteen years later, lyprinol, a nutraceutical product containing pure marine oil of the combination between lipid groups and unique Omega-3 polyunsaturated fatty acids had been reported by the Royal Melbourne Institute of Technology University. Interestingly, lyprinol was not only able to overcome the oxidation problem, but also showed a preeminent efficiency in acting against arthritis and other inflammatory conditions. Lyrinol was commercially available in 1990s. The extraction of lyprinol from Perna Canaliculus is a carefully prepared process from beginning to end. The shellfish used for providing mussels must be nourished in an ideal environment zone, which has pristine and uncontaminated water and rich of sunlight for phytoplankton growth, main food of the shellfish for growing mussels. Harvesting of mussels should happen in spring for optimal fatty acid yield (Daniell, 2006) and start in the early morning until mid afternoon and after that, they must be immediately transfered to processing plant, where they have further steps before the lyprinol extraction such as stabilization, lyophilising, grinding and packing. Later then, lyprinol is isolated from stabilized GLM powder through a patented supercritical fluid extraction process.

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Fig. Perna Canaliculus (left), lyprinol extraction room (middle) and commercial products of lyprnol (right).

Currenly, for higher yield of lyprinol extraction, there are many suggested innovations in aquaculture such as catching of spat in deep-water zones, offshore mussel farming, breeding selection, development of other environments than New Zealand for the sellfish growth. Biofuels: Recently, the study of green biofuel production from marine microalgae has been under investigation, which gives high promises of providing a new fuel that is clean, scalable and less effective on human health and enviroment. The aim of using the algae is to isolate the oil formed from algal metabolism and use it as source of supplying energy for transportation and home heating. In 1978, the Carter Administration had set up the Aquatic Species Program to investigate the ability of growing high-oil types of algae for biodiesel production. Interestingly, the result showed that algae farms could provide enough biodiesel, which could completely replace the use of fossil oil. In recent years, soladiesel, the first algal Fig. A Mercedes C320 powered by Solazyme fuel in an experiment. biodiesel has been introduced and demonstrated for its high effect on a factory-standard

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automobile for long distances under typical driving conditions (Paul, 2008) and its desire properties such as the maintenance of its liquid form in cold climate (Bee, 2008). Today, the agal biodiesel soladiesel still has to be under investigation to meet some other requirments before it can be widely used, thus, able to replace the use of petrofuel completely. Generally, farming algae in open pond systems using seawater and sunlight could achieve very high yield of oil. When a sufficient masss of algae reaches the requirments, they will be harvested by centrifugation or filtration,

dewatered, dried by heat or a vacuum and extracted oil by cell disruption using chemicals such as hexane or physical disruption such as ultrasonic sound waves. The amount of obtained algal oil could reach thousand gallons per acre per year, which is very higher than that of corn oil or palm oil production. However, due to the risk of contamination and water evaporation problem, farming algae in pond has been replaced by many new systems, which help overcome the problems and increase the output. The Vertigro Bio Reactor System is a typical example. In this system, the algae was cultured within plastic bubbles hanging vertically to enhance the exposure of algae cells to the sunlight. During 90 days of a repeated production test, algae were being harvested at one gram (dry weight) per liter, which is supposed to be equal to 33,000 gallons of algae oil per acre per year. In addition, it is necessary that we should mention about the technique that is developed by Solazyme Inc., company that produces soladiesel. The company, in order to decrease the production cost of making fuel but still obtain an adequate commercial level, has developed a new method in which the genetically modified strains of algae are cultured under heterotrophic enviroment. This means that instead of growing algae in ponds or place exposed to the sun, the algae are put inside stainless-steel tanks where light cannot occur. By modifying the algae, they are able to survive in the dark and are sustained by sugar. The sugar fed on algae will be converted into various types of oil, which will be isolated and processed to make a variety of beneficial biofuels.
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Fig. Algae farms using an open pond system

Fig. General Vertigro BioReactor System (left) and Glent Kerz, Chief Excutive Officer of Valcent Products, Inc. is standing by many algae bioreactors.

Fig. An illustration of soladiesel production process: during the fermentation, the materials providing sugars (sugarcane, sugar beets, wood wastes, etc) is put in the bioreactor, modified algae that no more require CO2 will use the supplied sugar to make oil as a way of storing its energy. High amount of oil is then extracted and utilized for soladiesel production.

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Environmental conservation: In recent centuries, there have been several reports about oil spill, a serious problem that could greatly endanger the living environment of millions of organisms and human life all over the world. The harmfulness of oil spill is due to its high toxicity formed form mixture of different types of oil, including crude oil, refined petroleum, and oil mixed with waste and its ability of remaining on water surface for a long time, which could kill numerous of organisms. Many approaches for cleaning up the oil spill have been made, including both chemical and mechanical manners. However, currently, the scientists are now focusing on biological method using marine microorganisms to clean up the oil spill on the ocean since it supposed to be less expensive, more effective and more friendly with environment, and by that, the marine biotechnogy has once again affirmed its important position in human life. In early 1940s, the American Petroleum Institute had been funding for researches studying the use of microbial systems to clean up oil spills. Later, more discoveries were obtained in the 1980s in which marine microorganisms were found to have the ability to degrade not only the petroleum products but also nearly every toxic Fig. Pseudomonas flourescens, which have been shown to be capable, degrades of oil and other hydrocarbons.

compound that enters the environment. Furthermore, a test using marine microbes to clean up the oil spill in Exxon Valdez

Alaskan was successfully made in 1989, which gave great perspective about the ability of cleaning up oil spill in more large-scale areas of pollution in near future. Currently, scienstists are trying to devolop new microorganism strains by genetically modifying them for the oil spill solution. This is because specific microorganism has specific structure for breaking down specific compounds, thus, it is hard to clean up a mixture of oil. Therefore, modifying a microrganism to make it digest multiple compounds that can appear in an oil spill is the best solution. Fortunately, Alcanivorax borkumensis, rod-shape bacteria that specify in
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aerobic decomposition and able to provide a base structure that can break down many types of hydrocarbons, therefore, they are supposed as the best candidate for the job. The method is carried out as the genes coding for hydrocarbons isolated from the biodegrading microorganisms are introduced into Alcanivorax through the DNA plasmids. The Alcanivorax after modifying will be test on a sample of mixed oils (alkanes, cycloalkanes, etc.) for the gene expression before they could be used for clean up oil spill in the environment.

Fig. Alcanivorax borkumensis and their desirabe properties for cleaning up oil spill.

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Reference
Pharmaceticals: http://www.mnhn.fr/publication/zoosyst/z00n2a15.pdf http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2992996/ http://resources.metapress.com/pdf-preview.axd?code=x235x2q6025055r4&size=largest http://www.esf.org/index.php?eID=tx_nawsecuredl&u=0&file=fileadmin/be_user/CEO_Unit/Sci ence_Policy/biotech.pdf&t=1283591772&hash=8f8f8c11032291dbe4d56d01889c71d2 http://www.sciencedirect.com/science?_ob=PdfExcerptURL&_imagekey=B8GY7-4P40D73-H1&_uoikey=B8GY7-4P40D73H&_piikey=S0165325398800151&_cdi=42459&_user=4861547&_acct=C000063557&_versio n=1&_userid=4861547&md5=8739d5aa0b14fe16fa1c8932745d308f&ie=/excerpt.pdf http://www.elsevierdirect.com/companions/9780120885305/casestudies/02-Ch27P088530web.pdf http://drug.pharmacy.psu.ac.th/wbfile/229254412362.pdf http://onlinelibrary.wiley.com/doi/10.1002/bit.20404/pdf http://nopr.niscair.res.in/bitstream/123456789/7759/1/IJBT%205(3)%20263-268.pdf Nutraceuticals: http://www.cancer.org/Treatment/TreatmentsandSideEffects/ComplementaryandAlternativeMedi cine/PharmacologicalandBiologicalTreatment/lyprinol https://www.lyprinolusa.com/faqs/#1 http://www.arthritistreatmentblog.com/lyprinol.html http://www.pharmahealth.co.nz/prd-lyprinol.php http://www.lyprinol.in/lyprinol-introduction/extraction/ http://www.business.auckland.ac.nz/LinkClick.aspx?fileticket=MO2H15Dv0hA%3D&tabid=22 85 http://www.lyprinol.ie/index.php?option=com_content&view=article&id=12&Itemid=29 http://www.lyprinol.ie/ http://www.green-lipped-mussel-oil.com/green-lipped-mussels-info.html Biofuels: http://biodieselreport.com/2008/01/24/solazyme-biodiesel-from-algae-soladiesel/ http://www.greencarcongress.com/2008/02/solazyme-ups-so.html http://www.solazyme.com/fuels http://www.ridelust.com/solazymes-algae-derived-biodiesel-passes-defense-departments-coldweather-testing/

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http://www.ridelust.com/solazymes-algae-derived-biodiesel-passes-defense-departments-coldweather-testing/ http://www.greenfuelsforecast.com/ArticleDetails.php?articleID=481 http://atlantabiofuels.websitetoolbox.com/post?id=2656174 http://www.rexresearch.com/kertz/kertz.htm http://www.renewableenergyworld.com/rea/news/article/2008/01/international-energy-developsmicroalgae-extraction-process-51299

http://www.scribemedia.org/2007/09/12/the-great-green-hope/ http://www.greentechmedia.com/articles/read/from-hype-to-reality-not-all-algae-were-createdequally/ http://sustainabledesignupdate.com/2008/02/

Environmental conservation:
http://saferenvironment.wordpress.com/2009/06/05/oil-spills-and-marine-biotechnology/ http://www.fas.org/ota/reports/9109.pdf http://www.marinebiotech.org/biorem.html http://www.research.noaa.gov/oceans/t_biotech.html

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