Marine Life, Including Fish, Seaweed, and Algae, Represents a Unique and Growing Global Resource for the Production of Nutraceuticals and Functional Food Ingredients

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Marine life, including fish, seaweed, and algae, represents a unique and growing global resource for the production of nutraceuticals and functional food ingredients. Photo copyright © iStockphoto.com/Tammy616 BY CHONG M. LEE, COLIN J. BARROW, SE-KWON KIM, KAZUO MIYASHITA, and FEREIDOON SHAHIDI

An expanding body of scientific research indicates that the marine environment is a unique resource of functional food ingredients with health-promoting properties.

esearchers may one day find in the ocean including cardiovascular disease risk reduction, immune “ answers to questions that will help to real- function improvement, brain health, and rheumatoid arthri- ize dreams of lengthening life and memory, of tis inflammation reduction. The oils are currently produced Rretarding aging, and of cures for a variety of ill- from fish, algae, krill, seal, and recently from squid. nesses,” stated Jacques Cousteau, who is considered Fish oil with omega-3 fatty acids (eicosapentaenoic a pioneer of ocean exploration (Cousteau, 1975). acid, EPA, and docosahexaenoic acid, DHA) experienced Much attention has been paid to exploration of poten- a sales increase of 35–40% in 2005–2006 (Hjaltason, tial nutraceuticals and pharmaceuticals derived from the 2007). The combined sale of consumer products forti- ocean. While the majority of nutraceutical products in fied with omega-3 fatty acids is estimated to be $19 billion the marketplace are of botanical origin, marine-based globally, according to the Global Organization for EPA and nutraceuticals are gaining attention due to their unique fea- DHA Omega-3 (GOED Omega-3) (Moloughney, 2011). tures, which are not found in terrestrial-based resources. Fish oil is produced from various sources, including A growing body of research indicates the potential for anchovy, menhaden, herring, mackerel, salmon, and cod the marine environment as a unique source of functional liver, and marketed in various forms, most commonly as food ingredients. In recent years, a series of promising concentrated omega-3 oil in soft gel capsules and microen- new marine nutraceutical products have been introduced capsulated powder. In order to improve yield and oxidative to the nutraceuticals and functional foods markets. stability, enzymatic concentration and microencapsula- This paper examines the current state of the marine tion of omega-3 fish oil have been carried out based on nutraceuticals and ingredients industry from an interna- a complex coacervation technology at Ocean Nutrition tional perspective, with a focus on the available marine Canada (Kralovec et al., 2009). The delivery of omega-3 bio-resources for nutraceutical products and functional oil in beverage products is being sought through a sophis- ingredients in global markets and their claimed health ben- ticated microemulsion technology (Moloughney, 2011). efits. Also discussed are emerging products, production Unlike other fish oil, farmed Atlantic salmon oil contains methods, ongoing R&D activities, and challenges in marine a 2:1 ratio of DHA and EPA, together with astaxanthin. nutraceuticals and ingredients markets in different parts The key producers of omega-3 oil are Ocean of the world, along with future developments in marine Nutrition Canada, Dartmouth, Nova Scotia, Canada; nutraceuticals and ingredients industry and research. Epax AS, Aalesund, Norway; and Napro Pharma AS, The main sources and products of primary inter- Brattvaag, Norway, from anchovy and sardine; and est for marine nutraceuticals and ingredients include Omega Protein, Houston, Texas, from menhaden. omega-3 fish/algal oil, phospholipids (bound omega Other producers include Nissui, Tokyo, Japan, and 3-fatty acids), micro/macro algal nutrition supple- Pronova BioPharma, Lysaker, Norway, for omega-3 oil- ments, fish proteins and peptides, hydrolysates, shellfish derived pharmaceuticals, and Croda, Goole, UK. chitin, fish collagen, and mineral supplements. Antarctic krill and squid oils contain mostly phospholipid-bound omega-3 fatty acids; krill oil also offers Omega-3 Polyunsaturated Oils a potent antioxidant, astaxanthin (Beer, 2011). Neptune Omega-3 polyunsaturated oils offer several health benefits, Technologies & Bioresources, Laval, Quebec, teamed with

12.11 • www.ift.org 23 pg Marine Life Sciences, Salt New entries to the algal oil market are Other than capsules and microen- Lake City, Utah, to mar- Algae Bioscience Corp., Overgaard, capsulated forms, omega-3 fish oil can ket a blend of Antarctic Ariz., (EPA and DHA) and Aurora be consumed in the form of micro- krill oil and concen- Algae, Hayward, Calif. (EPA only). emulsion in beverage products, oily trated marine algae Recently, Solazyme Roquette fish such as mackerel and salmon, for dual benefits. Aker Nutritionals, South San Francisco, or processed seafood such as nug- BioMarine, Oslo, Calif., has developed an egg and but- get, patty, and sausage formulated Norway, the world ter replacement ingredient, algal flour, from high omega-3 oil-containing fish largest krill oil pro- from microalgae for fat reduction. as well as omega-3 oil added surimi ducer, is the exclusive Mussel oil produced from New products, if fatty acids can be ade- supplier for Valensa Zealand green-lipped mussel (Perna quately protected from oxidation (Lee International, Eustis, canaliculus) contains eicosatetraenoic et al., 2007; Tolasa et al., 2010). Fla., which markets a acid (ETA 20:4n-3) and octacosa- krill oil-based supple- octaenoic acid (OOA: 28:8n-3) in Microalgae of Commercial Importance ment for joint and eye addition to EPA and DHA. It is used A commercial, large-scale microalgae health. The two compa- for inflammatory arthritis relief. culture started in the early 1960s in nies are also performing Supercritical fluid extraction (SFE) Japan with the culture of Chlorella. In

joint clinical studies of based on pressurized CO2 (liquid the following 40 years, the microal- krill oil. Enzymotec, at 31°C and 7.38 MPa) is used in gal industry has grown and diversified Migdal, Israel, is mussel oil extraction. The bioac- significantly. Currently, the micro- another global supplier tive marker for anti-inflammatory algal biomass market produces about Each 240-ml serving of Stonyfield Organic of refined krill oil. activity has not been isolated, but 5,000 tons of dry matter/year and Whole Milk provides 50 g of omega-3 Parma Marine, such activity is believed to be from generates a turnover of approximately EPA/DHA for heart, brain, and eye health. Ocean Nutrition Canada supplies Terøy, Norway, mar- the synergetic function of various U.S.$1.25 billion/year (Pulz and the omega-3 ingredients, which are kets phospholipid-bound omega-3 fatty acids (EPA, DHA, and Gross, 2004; Spolaore et al., 2006). derived from fish oil. omega-3 fatty acid-con- ETA) (Sankaran and Mouly, 2007) Chlorella (Chlorella vulgaris and taining oil from squid along with glycosaminoglycans. Chlorella pyrenoidosa) is rich in chlo- byproducts. Phospholipids of marine Seal oil, which contains DHA, rophyll, protein (40–60%), and origin are a good source of more- EPA, and docosapentaenoic acid carotenoids. It exhibits putative bioavailable omega-3 fatty acids, (DPA: 22:5n-3, clupanodonic acid), is anti-carcinogenic, immunomod- particularly for brain health. These anti-atherogenic (Nakhla, 1998) and ulatory, hypolipidemic, gastric phospholipids have been used for is more stable than fish oil (Shahidi, mucosal-protective, and detoxi- memory treatments (Louis-Sylvestre, 1994). It is currently produced in fication activities. Producers of 1999) and are an important ingre- Newfoundland and Labrador, Canada. Chlorella include Cyanotech, Kailua- dient in larval fish feed (Lian et al., Shark liver oil contains squalamine Kona, Hawaii, and Earthrise 2008). Currently, marine phospholip- (aminosterols) and squalene (alkyl- Nutritionals, Ishigaki Island, Japan. ids are produced from krill and squid glycerols), which deliver potential Spirulina (Spirulina platensis) is one in commercial quantities. immune enhancement, anti-angio- of the few plant sources of vitamin Algal oil with DHA is cur- genic activity, and skin conditioning B12. It provides immune enhance- rently produced by DSM-Martek (Newman and Cragg, 2004). ment, protein supplementation Biosciences, Columbia, Md., Industry analysts estimated (65–71% by weight), and has puta- through heterotrophic culture of global consumption of marine and tive health benefits including anemia Crypthecodinium cohnii (DHA > 30%) algal omega-3 ingredients in 2008 prevention, hypocholesterolemic, for infant nutrition formula in com- at 71,452 tons, with North America antioxidant, hepatoprotective, and bination with fungal arachidonic acid being the largest consumer at 26,948 anti-allergic activities. Producers of (ARA) and Schizochytrium sp. (DHA tons, followed by Asia-Pacific at Spirulina include Cyanotech; Earthrise > 33% with 13.5% docosapentae- 21,145 tons, European Union (EU) Nutritionals, Irvine, Calif.; and Parry noic acid, DPA: 22:5n-6, osbond acid) at 13,596 tons, and the remainder of Nutraceuticals, Chennai, India. for general nutrition supplements. the world at 5,762 tons (Real, 2009). Dunaliella salina is a primary Lonza Group, Basel, Switzerland, Currently, there is about 85,000 algal source of ß-carotene (antioxi- produces microalgal oil (DHA > tons, according to GOED Omega-3 dant) with putative aging retardant 43%) from Ulkenia sp. (Kiy et al., (Moloughney, 2011). In an effort and immune enhancement. It is capa- 2005). Photonz Corp., Auckland, to implement quality standards and ble of producing bioglycerol. Cognis New Zealand, has developed phar- also to increase consumer aware- Nutrition and Health, Tullamarine, maceutical-grade EPA omega-3 oil ness and regulatory approval, GOED Australia (now part of BASF) manu- from cultured diatom Nitzschia laevis. Omega-3 was established in 2006. factures and markets Dunaliella salina. »» pg 24 12.11 • www.ift.org Crypthecodinium cohnii pro- enhancement, type 2 diabetes control, duces DHA (> 30%), which is anti-hypertensive (ACE inhibition), being used primarily for infant for- anti-inflammatory, and anti-can- mula with arachidonic acid (ARA). cer activities. In addition, seaweed DSM-Martek Biosciences is a sup- may provide UV-protective func- plier of Crypthecodinium cohnii. tion, natural sunscreen, and skin care Schizochytrium sp. produces potential (Lee and Apostolidis, 2009). DHA (> 35%) plus 13.5% DPA Potential bioactive components and is used as a nutritional supple- in seaweeds responsible for nutra- ment and active ingredient in poultry ceutical activities include fucoidan feed. DSM-Martek Biosciences is (sulfated polysaccharide), fuco- a supplier of Schizochytrium sp. xanthin (carotenoid), fucosterol, Nitzschia laevis produces EPA. phloroglucinol-based polyphenols Photonz Corp. has developed a phar- (phlorotannins and simple pheno- lics), glycoproteins, and peptides. maceutical-grade EPA omega-3 Microalgae cultures are grown at the Commonwealth Scientific and Industrial manufacturing process from micro- Fucoidans are sulfated polysaccha- Research Organization (CSIRO) Marine Research in Australia to maintain an algal biomass along with SFE rides (> 40% of the dry weight of the active collection for both scientific and commercial use. technology from Separex S.A., algal cell walls) and have demonstrated Photo courtesy of CSIRO, Hobart, Australia Champigneulles, France. Its applica- beneficial effects on cardiovascu- tion is aimed at cardiovascular disease. lar, joint, and digestive systems in an enzymatic hydrolysis of chitosan using Haematococcus pluvialis produces experimental studies. They are also chitosanase (Kim and Rajapakse, 2005). astaxanthin (7% of dried weight), known to boost the immune system, It may offer several benefits, includ- a dietary antioxidant (500x vita- help protect against thyroid cancer ing binding excess fat and inhibiting fat min E). Producers of Haematococcus and other cancers (Itoh et al., 1993; absorption, lowering LDL, boosting pluvialis include Cyanotech; Mera Zhuang et al., 1995), moderate blood HDL cholesterol, anti-cancer, antimicro- Pharmaceuticals, Kailua Kona, sugar and cholesterol levels (Murata et bial, immune enhancement, lowering Hawaii; Valensa International; Parry al., 1999), and detoxify the body from blood sugar, controlling blood pressure, Nutraceuticals; Astareal, Gustavsberg, heavy metals (Morris et al., 1999) enhanced absorption of calcium, reduced Sweden; and Algatechnologies, and radioactive elements (Carlson and risk of cardiovascular disease, and Kibbutz Ketura, Israel. Erlandsson, 1991). A comprehensive reduced level of uric acid in the blood, review discussed the therapeutic role thus preventing the occurrence of gout. Seaweeds of fucoidan in the control of acute and In recent years, seaweed has gained chronic inflammation (Fitton, 2011). Other Sources of Marine Nutraceuticals much attention due to interest in its Fucoxanthin, a carotenoid with Bioactive peptides from hydrolyzed nutraceutical potential and develop- formula C42H58O6, is reported to bonito (Sarda orientalis) show anti-ACE ment of seaweed-based functional promote fat burning within fat cells (angiotensin-I-converting enzyme). foods and nutraceuticals world- (thermogenic) by increasing the ACE is a key enzyme in the regulation wide, the most actively in China, expression of thermogenin (uncoupling of peripheral blood pressure which Japan, and Korea. For instance, protein 1, UCP1) (Miyashita, 2009). converts angiotensin I into angioten- Marine Bioprocess Co., Busan, Fucosterol has demonstrated sin II, a highly potent vasoconstrictor Korea, focuses exclusively on sea- hypocholesterolemic activity, anti- molecule. This enzyme causes a reac- weed-derived functional drinks, oxidant activity (Lee et al., 2003), tion that increases both the volume foods, and cosmetics. The company and anti-diabetic activity (Lee et of blood flowing through arteries employs fermentation technology. al., 2004) when brown seaweed and the degree of constriction of the There are several known seaweeds (Pelvetia siliquosa) was tested. blood vessels. These peptides lower that offer nutraceutical properties Phloroglucinol-based polyphenols blood pressure by inhibiting ACE and can be developed into functional (phlorotannins) have exhibited anti- (Fujita et al., 2001) and help maintain foods. They include Laminaria japonica, oxidant activity (Smith and McLean, healthy blood pressure. Commercial Undaria pinnatifida, Ecklonia cava, Fucus 2006; Hwang et al., 2010), inhibi- products made from hydrolyzed ceranoides, Ascophyllum nodosum, Ulva tory effect on α-glucosidase and bonito include Levenorm from Ocean lactuca, Carpophyllum flexuosum, and α-amylase linked to type 2 diabe- Nutrition Canada, PeptACE from Cladosiphon sp. Seaweed may possess tes (Apostolidis et al., 2008), and Natural Factors, and Bonito Peptide nutraceutical and cosmeceutical prop- UV-protective function as a potential from Ortho Molecular Products. erties, including antioxidant, fat cell natural sunscreen (Heo et al., 2009). Fish protein hydrolysate produced thermogenesis, retarding fat absorp- Chitosan oligosaccharide is an oligo- from lean white fish through microbial tion, hypocholesterolemic, immune mer of D-glucosamine and obtained by fermentation, a technology developed

12.11 • www.ift.org 27 pg for dry, mature, or sun-damaged bioactive, pharmaceutical, and skin. It was developed by Ferrosan, nutraceutical products. a Danish healthcare company, which uses a patented fish extract Industry and R&D that contains properties simi- Ocean Nutrition Canada is in the lar to those found in the dermis business of discovery, manufactur- (notably collagen and elastin). ing, and marketing marine-based The development of a salmon- ingredients, which improve human based skin cream has been aided by health and are used for dietary sup- researchers at Norway’s University plements and functional foods. It is of Science and Technology, who dis- a world-leading producer of con- covered the skin softening enzyme, centrated omega-3 fish oil, along zonase, from the hatching fluid of the with microencapsulated omega-3 salmon eggs. Zonase helps dead skin oil powder as a food ingredient for flake off and stimulates the growth of functional food and dietary supple- healthy, new skin cells. It has also been ment applications. Production and proven helpful in healing wounds. The delivery technologies for omega-3 skin cream is sold under the brand fatty acid concentrates have also been name Aquabeautine XL from Aqua developed (Kralovec et al., 2009). Bio Technology, Bergen, Norway. Copalis, Portel, France, is a global Ascophyllum nodosum is one species of seaweed/algae that offers health- Marine nutraceuticals-related supplier of marine-based natural promoting properties for use in functional foods. research is being conducted in the ingredients for nutraceutical, func- Photo copyright © iStockphoto.com/LesleyJacques U.S. at various marine biotechnol- tional food, animal nutrition, and skin ogy research centers. They include care products utilizing fish process- by Proper Nutrition, Reading, Pa., Marine Biotechnology Center (MBC) ing byproducts for marine bioactives is shown to promote intestinal health (Univ. of California-Santa Barbara); through hydrolysis and fractionation. and maintain healthy bowel function. Center for Marine Biotechnology It formed a research alliance with Commercial products containing fish and Biomedicine (CMBB)/Scripps Ifremer (French Research Institute protein hydrolysate include Seacure Institution of Oceanography (Univ. for Exploitation of the Sea), Aquimer and Intestive from Proper Nutrition. of California-San Diego); Center (National cluster for aquatic prod- Collagen for skin care has been of Marine Biotechnology (COMB) ucts), Nausicaä (The French National produced mainly from cold water (Univ. of Maryland-Baltimore); The Sea Experience Centre), Nutrition fish skins such as cod, haddock, and Center of Excellence in Biomedical Healthcare Longevity Cluster, and salmon. However, recently, colla- and Marine Biotechnology (Harbor SEAFOODplus. SEAFOODplus is an gen is being commercially produced Branch Oceanographic and Florida integrated seafood research program from fish scale, such as tilapia, by Atlantic University); Marine supported by the European Union. decalcification and enzymatic hydro- Bioproducts Engineering Center NutraMara (Marine Functional lysis (Nagai et al., 2004). The (MarBEC), Hawaii-Manoa; and Food Research Initiative), Dublin, uniqueness of scale collagen is its recently Food Science and Nutrition Ireland, focuses on fish process- molecular weight, which is lower Research Center (Univ. of Rhode ing waste streams, underutilized (~ 1,000) than skin collagen. Island, Kingston). Their common raw materials, and underutilized Since 1991, Imedeen has been a interests are in marine biotechnol- species of fish and seaweed with leading internal skin care product ogy and marine organisms-derived an interest in the development of value-added marine functional Figure 1. Continuous separation of bioactives using an ultrafiltration membrane reactor system. foods while fostering collaboration Courtesy of Marine Bioprocess Research Center, Pukyung National University, Busan, Korea between higher education research institutions and the food and pharmaceutical industries. “Our seas raw material are a huge reservoir for bioactive enzymes compounds that can be incorpo- rated into food additives, which can bioreactor products UF membrane filtration be harnessed for human health,” said Maria Hayes, Scientific Project Manager of the NutraMara project. bioactives

pg 28 12.11 • www.ift.org Bioconversion of Marine Biomaterials using fractionation techniques such as ultra- and Processing Byproducts filtration (UF) membrane for continuous Bioconversion is a process by which raw mate- operation (Jeon and Kim, 2000; Park et al., rials or substrates are converted into useful, 2008) (Figure 1) or fractionation by centrif- value-added products by biological means, most ugation at selected g forces (Lee, 2009). commonly done with aid of endogenous or exog- Production of bioactives by fermentation enous enzymes. On a global scale, directly with probiotic cultures was done by fermenting accessible marine biomass represents up to 100 million tons of fish/year, crustacean and mol- lusks from fisheries, and aquaculture. It is estimated that more than 50% becomes waste or is underutilized (European Science Foundation Marine Board, 2001). This poorly utilized biomass is an important source of biologically active molecules possessing unusual properties. Various research projects are being actively carried out under SEAFOODplus, an integrated umbrella research program supported by the EU. One of the main objectives is to screen, map, and recover new, health beneficial compounds from seafood byproducts and underutilized species by advanced bioconversion processes in order to develop new bioactive, functional seafood ingredients (www.seafoodplus.org). According to SEAFOODplus studies, all fish hydrolysates exerted a weak to moderate in vitro inhibition of angiotensin-I-converting enzyme (ACE), suggesting that hydrolysis of fish proteins yields hypotensive peptides. Fish protein hydrolysates exhibited CT (calcitonin)-like activities for calcium and phosphorus metabolism for the treatment of hypocalcaemia or osteoporo- sis; and gastrin and CCK (cholecystokinin)-like activities for stimulation of digestive enzyme secretion, which restrict stomach empty- ing, thus giving a false sense of fullness. Gastrin stimulates secretion of gastric acid, while CCK causes the release of digestive enzymes and bile to stimulate the digestion of fat and protein. Hydrolysate peptides also exhibit antioxi- dative and free radical scavenging properties. Matis (Iceland Food and Biotechnology Research) Reykjavík, Iceland, employs alkaline protein recovery prior to enzymatic hydrolysis for production of superior quality peptides. Marine Bioproducts, Storebø, Norway, is a production and research company focusing on gentle and natural processing of fresh raw materials of marine origin. Enzymatic hydrolysis at low temperatures is the basic concept, followed by separation steps that yield high-quality protein-, lipid-, and bone- derived products. The current product line includes hydrolysates, peptides, amino acids, digested protein meals, minerals, and oil from salmon. Products yielded by bioconversion need to be further refined to separate target bioactives

12.11 • www.ift.org 29 pg squid processing byproduct with L. rhamnosus Marine Nutraceuticals Today & Tomorrow and P. acidilactici, yielding good DPPH scaveng- Marine nutraceuticals take a small share of the ing activities and IL-6 and TNF-α activities $22 billion global nutraceuticals market projected (Xu et al., 2009). A similar process was used in 2013 (Moloughney, 2009). Global interests in by Proper Nutrition to produce fish pro- marine bio-resources have spurred several interna- tein hydrolysate for intestinal health. tional conferences, including “Marine Ingredients Conference,” held in Oslo, Norway in September 2010, and “Biomarine Business Convention,” held in Nantes/Saint-Nazaire, France this past September. The marine nutraceuticals market will grow steadily as the global nutraceuticals and functional foods markets continue to expand. Aging Baby Boomers will help enlarge the marine nutraceuticals market. Developers and marketers of marine nutraceuticals and bioingredients should focus on identifying products for domestic or global markets; exploring indigenous bio-resources for domestic markets, building on the existing infra- structure, and expanding to overseas markets; securing sustainable resources through aquaculture and cell culture; and establishing high-throughput screening tests for candidates based on biomark- ers and nutrigenomic approaches (Lee, 2009). Development of marine nutraceutical products and ingredients is highly market driven and requires good marketing strategies. Special attention should be given to types of products (i.e., marine nutraceutical ingredients or functional foods) and to the most suitable form of product in terms of compatibility, stability, consumer acceptability, and regional preference. To succeed in the marketplace, products require good science-based clinical evidence. Marine omega-3 oil will enjoy continuous growth, while algae-based products will find expanding opportunities. FT

Chong M. Lee, Ph.D., a member of IFT, is Professor, FSN Research Center, Dept. of Nutrition and Food Sciences, University of Rhode Island, Kingston, RI 02881 ([email protected]), Colin J. Barrow is Professor and Chair, Dept. of Biotechnology, School of Life and Environmental Sciences, Deakin University, Geelong, VIC 3217, Australia ([email protected]), Se-Kwon Kim is Professor, Marine Bioprocess Research Center, Dept. of Chemistry, Pukyong National University, Busan 608-737, Republic of Korea (skn- [email protected]), Kazuo Miyashita, a member of IFT, is Professor of Bio-functional Material Chemistry, Faculty of Fisheries Sciences, Hokkaido University, Hakodate 041-8611, Japan ([email protected] dai.ac.jp), and Fereidoon Shahidi, a member of IFT, is University Research Professor, Dept. of Biochemistry, Memorial University of Newfoundland, St. John’s, NL, Canada A1B 3X9 ([email protected]).

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