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`
`Petition for Inter Panes Review
`Of U.S. Patent 8,278,351
`Exhibit
`
`ENZYMOTEC - 1016
`
`AKER877|TC00118581
`
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`Downloadedby[JohnJones]at18:5312Septentber2011
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`Food Reviews International, 6(1), 119-147 (1990)
`
`[Hi-. U I ILILAI ION Oi‘ AN IARLI IL KRILL
`
`FOR HUMAN FOOD
`
`TANEKO SUZUKI
`Hilton Universitv
`
`Department of Agriculture and Veterinary Medicine
`Kameino-i866. Fujisawa-shi, Kanagawa Prefecture, Japan
`
`NOBUXAZU SHIBA TA
`National Research Institute of Fisheries Science
`Fisheries Agency, Ministry of Agriculture and Forestry
`5—5-I Kacltidol-ti, Chuo~‘ku, 104 Tokyo, Japan
`
`ABSTRACT
`
`Antarctic krill {Euphauria superba) is distributed south of 60 "3
`around the South Pole. The stock of krill is estimated at 360 to [400
`lllllflll tons. In-1930 the total a ount harvested in the world was 500
`
`thousand tons, mainly by the USSR, followed by Japan. Chemical
`composition of krill is as follows: moisture 'l'7.9~83.l%, crude pro-
`ein ll.9~l5.4%, chitin and glucides 2%, and crude ash 3%. Nutri-
`tive value of krill protein is lower than whole—egg protein but higher
`han milk protein. Krill contains large an l0LlI'||'.S of vita
`tins A and B.
`About 10% of krill lipid is unsaturated fatty acids such as olcic, eico~
`sapentacnoic acid, and docosahexaenoic acid. Commercial products
`from krill in Japan are frozen raw krill, frozen boiled krill, peeled
`krill meat, and others. All of these products are processed on boats
`in the Antarctic Ocean. Krill products. in Japan totaled 2'll,O50 tons
`in l9Bfi~l98?.
`
`Copyright © 1990 by Marcel Dekker, Inc.
`
`119
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`000002
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`AKER877|TC00118582
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`000002
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`Downloadedby[JohnJones]at13:53l2September20.11
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`1211
`
`SUZU Kl AND SHIBATA
`
`Kl-KILL BIOLOGY AND
`
`FISHERIES
`
`Taxonomy of Krill
`
`“Krill” is the general name for the small, shrimp-like organisms belonging to
`the order Euphausiacae, which are found in both shallow and deep waters around
`the earth, particularly in areas of high latitude. “here are 2 families, I] genera,
`and 84 recognized species belonging to krill (1, 2). Among them the Antarctic
`krill (Euphausia superba) is the ‘nost abundant and the -nost important. Its utili-
`zation as a potential resource for human food has called much attention to it,
`althot gh there are a few other species living in the Antartic Ocean besides E.
`superba.
`The taxonomy of E. superba is as follows:
`
`Phylu 11
`
`Arthroooda
`
`Subohylum
`Class
`
`Mandibulata
`
`Crustacca
`
`Subclass
`
`Series
`
`Malacostracea
`
`Eumalacostraca
`
`Suoerorder
`
`Eucardia
`
`Order
`
`Family
`Genus
`
`Euphausiacea
`
`Euphausiidae
`
`Euphausia
`
`Species
`
`Euphausia saperba
`
`The E. superba grows as
`smaller.
`
`large as about 5 cm. Most of the euohausiid are much
`
`is similar to that of shrimp belonging to the order of
`The apoearance of kr'll
`Decaooda of the same superorder Eucarida (Fig. 1). Smaller sine and exposed
`gil are points to generally disthguish Euphausiacea from Decapodae. Another
`difference is that the forner spends its whole life floating as plankton, whereas
`the latter, in most species, floats only in the lava stage.
`"‘he Antarctic krill is distributed south of 60"S around the South Pole, with
`
`high density in the cold waters of low salinity (Fig. 2}.
`During daylight hours, schools of the Antarctic krill are found mainly in depths
`of S0 to 100 in, while in the evenings they float up to the surface and are fre-
`quently seen as brownish “patches” (3).
`The large variety of food for krill includes plant and animal detritus material;
`and at the same time, krill itself is the main food for whales, seals, fishes, and
`birds. Thus, it "Jlays an important role in the food chain system in the Antarctic
`ecosystem.
`
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`Imam I. Autarmia km mm: mm: :s5:m»::ie:; m‘ km: um) Aniartms mm; (b)?a1‘n hummer krill (fiuéw
`phnumh xwpewaajg seam, krill (Euphawia pamjfica); and tight, prawn {Sergmrwm !%rW£3£§»\W3‘}.'
`
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`122
`
`SUSEUKI AMI.) 2‘”viHmA'I‘A
`
`Friggurt 2.. Distrihmsm M km! in Anmrcztisx Ocean.
`
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`Downloadedby[JohnJones]at18:5312September2011
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`UTILIZATIDN OF ANTARCTIC KRILL
`
`123
`
`Stock Size of Krill
`
`Although the Antarctic Ocean has very high biological activity, and krill sus~
`tains various kinds of animals, the stock size of krill rerrains very considerable.
`In the years when whales in the Antarctic were not exploited, it is estimated that
`more than 180 million tons of krill were COHSUI ed by baleen whales, and over
`300 million tons were CUHSUI ed by other animals such as seals, fishes, birds,
`etc. (4). ln most cases, the estimate of fish resources is rrade by analyzing fish
`catch statistics. in the case of krill, however, this method is insufficient because
`krill catch data do not cover the entire stock of krill. Therefore, several scientists
`have tried sorre indirect 1 ethods to estirrate the krill stock size by an analysis
`
`of the dynamics of the ecosystem. Their methods vary, but in general, a numeri-
`cal model is set up and so ved by using biological parameters such as reproduc—
`tivity and growth of krill, those of krill feeders such as whales and seals, con-
`version ratio between prey and predator, and so on.
`e result varies rather widely
`according to individual scier tists because of the different assumptions introduced
`in setting up the model of each type and because of the lack of knowledge of
`reliable fundar ental par
`ieters. The estimated value ranges frorr 350 million
`to about a billion, to about 700 to 1,400 million tons (5, 6).
`The rrethod of estimating stock size by using an echo~integrator, or “scienti-
`fic fish-finder," has been developed in recent years. In this method, neither any
`assumed biological parameter nor any catch data are used in the calculations.
`The estimate of the stock of krill is likely to be low because the equipment can
`
`measure only to the depth of 100 m, and rrany schools of high density have been
`found in deeper waters. The stock size of krill in the entire Antarctic Ocean can
`be obtained from the local density measured by this method. An example in such
`an estimation is about 300 million tons, with a high possibility of underestimated
`bias from the viewpoint of: athematical statistics (7). Notwithstanding such a
`defect, th's method is extrer ely useful in studying the local distribution pattern
`of krill in relation to the env‘ronr ental conditions; therefore, further develop-
`ment of the technology is expected in both hard and software aspects.
`The 3! ount of catch per hauling, which represents the local density of the
`stock, stays stable except for the early period of exploration (5) (Fig. 3). Despite
`the present world catch of Antarctic krill of 0.3~0.5 r
`illion tons, there seems
`to be no direct impact of the catch upon the stock. At the same time, it should
`be mentioned that the distribution of krill is far fro:
`ever . Therefore, an ex-
`
`ight cause a local overcatch
`treme corcentration of effort in any single locale r
`of local stock, resulting in the lowering of catch per effort. Fish'ng for krill under
`such circumstances becomes unprofitable.
`
`Fisheries
`
`Exploitation of Antarctic krill started in the early 19605 by both Japan and USSR
`for research purposes. Now in the 19805, nine countries have krill fisheries. In
`
`000006
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`AKER877|TC00118586
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`124
`
`SUZUKI AND SHIBATA
`
`10
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`
`Figure 3. Amounts of catch of krill per one net haul. Source: Nasu 1988.
`
`1913 the total harvest in the world was 22 thousand tons; almost all was by the
`USSR. In the early 198% the world catch was as large as 500 thousand tons, mostly
`by the USSR, followed by Japan. Since then, there has been a sudden drop in
`the USSR catch, and the total world catch decreased in spite of the increase of
`the Japanese catch in recent years for research purposes (Fig. 4).
`The majority of the Japanese catch comes from the Atlantic Sector, in par-
`ticular from the Scotia Sea despite its distance from Japan. One reason for this
`is that the standing crop of krill in this area is more stable than in other areas.
`Japanese krill fisheries are operated by means of stern trawl vessels with sur-
`face or mldwater trawl nets.
`
`The inactivity of recent krill fisheries is not due to the decline of stock but
`completely due to the stagnation of the market and inbalanee between cost and
`price. If improvements are made to increase economical usage of krill. fisheries
`can increase their catches several times the present level without fear of hav-
`ing an adverse effect on krill stock or on-the balance of the total Antarctic eco-
`system. Figure 5 shows the Japanese fisheries in the Antarctic Ocean.
`
`CHEMICAL COMPOSITION OF Al"llTARCTIC KRILL
`
`Chemical Co: positio i Ge eral
`
`A number of reports have been published by scientists of Japan, USSR, and
`other countries. The reports differ slightly because of the differences in size, age,
`
`000007
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`Downloadedby[JohnJones]at18:5312September2011
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`Downloadedby[JohnJones}at18:5312September2011
`
`126
`
`SUZUKI AND SHIBATA
`
`sex of samples, and seasons of sampling. Table l (B) shows the chemical compo-
`sition of whole krill (with earapaces) taken in the period December to February.
`Size and chemical composition of krill vary from season to season. The amount
`of water decreases, and protein and fat increase with age of krill. The average
`analyses are: T19-83.1% water, ll.9—l5.4 "Zn crude protein, about 2% of chitin
`and glueides, and about 3% ash (9). The composition of muscle after removing
`cephalothorax and carapace is shown in Table 2. It contains, naturally, less ash
`as compared to whole krill. Moreover, no remarkable difference can be seen be—
`tween the cephalothorax and teison, Table 3 (13).
`
`Table 1. Variation of Chemical Composition of Whole Krill
`
`Dale of catch
`Dec. 5
`Dec. 55
`Dec. 26
`Jan. 5
`Jan. I5
`Jan. 25
`Feb. 5
`
`Feb. I5
`
`Moisture
`{"211}
`33.7
`31.3
`31.2
`80.4
`80.3
`79.7
`73.8
`
`30.0
`
`Crude protein
`{"24}
`11.3
`13.8
`13.?
`14.2
`13.5
`14.2
`14.2
`
`13.4
`
`Crude fat
`, {We}
`0.48
`0.60
`0.77
`L48
`1.90
`2.50
`3.6]
`
`3.3?
`
`Glucide
`(We)
`0.62
`0.66
`0.73
`0-55
`0.56
`0.53
`0.60
`
`0.59
`
`Chitin
`{We}
`0.53
`0.53
`0.52
`0.50
`0.52
`0.50
`0.49
`
`0.50
`
`Crude ash
`(We)
`3.24
`3.l9
`2.83
`3.00
`3.07
`3.0?
`3.08
`
`2.60
`
`Table 2. General Components of Antarctic Krill Meat
`
`General component (070)
`
`Sample no.’ Water
`
`Crude protein
`
`Crude fat
`
`Crude ash
`
`Researcher and Ref.
`
`I
`2
`3
`
`4
`
`76.50
`79.95
`79.69
`
`34.2
`
`19.63
`1165
`1?.-41
`
`12.9
`
`2.65
`L31
`1.54
`
`0.4
`
`L43
`1.43
`1.45
`
`Ll
`
`Hirano at al. {[0}
`Suynrna et til. (I 1)
`Suyaina cl nl. (ll)
`
`Suzuki et al. (12)
`
`"Samples l, 3: frozen; Sample 2 collected from stomach of Antarctic whale; Sample 4 water content very
`high because of the large quantity of water used when shell is removed by pee.-ling machine.
`
`Table 3. Difference in Che
`
`ical Components of Ceplialothorax and Tail of Krill
`
`Part of
`krill body
`
`Cepltalothorax
`Tail
`
`”"
`Water
`
`'."9.7
`79.0
`
`General component {Wu}
`
`Crude protein
`
`Crude fat
`
`Crude ash
`
`Weight ratio
`in whole body (Wu)
`
`l2.'!
`14.6
`
`2.6
`2.2
`
`4.4
`3.2
`
`45
`55
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`Downloatledby{JohnJones]at18:5312September2011
`
`UTILIZATIGI‘-I (IF ANTARCTIC KRILL
`
`12'?
`
`The chemical composition of krill is characterized by a little more moisture
`and more glucides than traditional" commercial lish. On the whole, krill has a
`well—balanced composition for human foods.
`
`Lipids, Vitamins, and Minerals
`
`The content of lipids in k'rill varies greatly and is determined by the stage of
`growth at the time of catch. The lipid content increases toward the end of the
`fishing season in the Antarctic Ocean (14).
`The content of phospholipids in krill was reported by Aral et al. (15) to be
`29.9%, and by Bottino {9} to be 54-58%. The lipids of krill have a high iodine
`value. as shown in Table 4, which indicates rather high unsaturation as com-
`pared to other fish species (16).
`Reports of fatty acid components of krill lipid have been made by many sci-
`entists (17-23}. On the basis of their reports, Grantha r (9) summed up the values
`as follows: About 70% of,the whole lipid was unsaturated fatty acid with an
`iodine value of 110-190. Concentrations of oleic acid, eicosapentaenoic acid,
`and docosahexaenoic acid are high. Essential Fatty acids such as linolic, linolenic,
`and arachidonic acids are 5% ofthe total lipids.
`Table 5 shows the fatty acid composition of krill lipids (14). Yanase (2%!) com-
`pared the fatty acid components of krill lipid with those of salmon, tuna, mack-
`erel, baleen whale, and shrimp. He reported that krill lipid closely resembles
`that of the baleen whale and also that of lish. Krill lipids are, however, not easily
`oxidized, which may be due to their high vitamin 13 concentrations (16, 25, 26).
`Krill lipids occasionally show a high acid value because frozen samples are
`stored for long periods. It is known that frozen fish containing high percentages
`of phospholipids have high acid contents (24)-
`
`Table 4. Unsaponifiable Matter and Cholesterol in Krill
`
`Total cholesterol
`
`Sample type“
`A
`A
`B
`B
`B
`
`B
`
`Iodine value
`l35
`166
`l6-0
`187
`I32
`
`I40
`
`Unsaponifiable matter
`(Cite in oil}
`4.68
`8.56
`3.5?
`13.1
`4.80
`
`4.48
`
`mg/g oil
`l9.l
`42.6
`48.0
`76.3
`2l.?-
`
`17.0
`
`“A: raw krill was frozen; B: boiled krill was frozen.
`
`igfllll 3;. tissue
`71.6
`63.l
`71.0
`62.3
`66.7
`
`62.1
`
`0000010
`
`AKER877|TC00118590
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`0000010
`
`

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`
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`
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`Downloadedby[JohnJones]at18:5312September2011
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`1211
`
`SUZUKI AND SHIBATA
`
`Table 5. Fatty Acid Composition of Krill Lipids ("79)
`
`Sample A”
`
`Sample 13"
`
`Fatty acid
`2:0
`3 :0
`4:0
`4:1
`15:0
`16:0br
`6:0
`6:1
`6:2
`2:0
`7:1
`3:0br
`8:0
`8:1
`18:2
`13:3
`3:4
`9:11
`9:1
`20:0
`20:1
`20:2
`20:3
`20:4
`20:5
`21 :0
`21:1
`22:1
`22:2
`22:3
`22:4{24:1)
`22:5
`22:6
`23:1
`
`24:4(?1
`Unknown
`
`Saturated
`
`“Monounsaturated
`Polyunsaturated
`Unknown
`
`Total
`lipid
`0.1
`Trace
`4.9
`0.2
`0.5
`0.2
`13.3
`4.9
`1.4
`0.1
`1.0
`0.1
`1.0
`16.4
`3.3
`1.1
`1.3
`0.2
`0.1
`0.2
`0.4
`0.2
`1.2
`0.5
`17.4
`Trace
`0.4
`0.4
`0.2
`5.3
`0.2
`0.7
`12.4
`0.4
`
`2.8
`1.2
`
`26.1
`
`24.2
`43.5
`1.2
`
`Neutral
`lipid
`0.1
`Trace
`6.9
`0.2
`0.6
`0.1
`17.9
`6.0
`2.0
`0.1
`1.1
`0.2
`0.9
`17.1
`3.2
`1.3
`1.7
`0.2
`0.1
`0.4
`0.4
`0.1
`1.5
`0.4
`17.6
`0. 1
`0.5
`0.4
`0.3
`4.5
`0.4
`0.4
`9.2
`0.2
`
`3.1
`0.8
`
`27 .5
`
`26.0
`45.7‘
`0.8
`
`Complex
`lipid
`0.1
`—-
`1.11
`0.1
`0.3
`0.1
`23.3
`2.7
`0.4
`Trace
`0.7
`0.2
`0.7
`15.0
`3.7
`1.2
`0.9
`0.2
`0.1
`0.2
`0.4
`0.3
`1.3
`0.3
`16.5
`Trace-.
`0.3
`0.2
`0.5
`4.9
`0.4
`0.6
`14.4
`0.6
`
`6.4
`1.2
`
`26.9
`
`20.1
`51.8
`1.2
`
`Total
`lipid
`0.2
`Tr act:
`11.1
`0.3
`0.9
`0.2
`17.6
`6.2
`1.7
`0.1
`0.9
`0.3
`0.9
`14.‘!
`3.4
`1.5
`3.1
`0.4
`0.1
`0.3
`0.1
`0.1
`1.4
`0.6
`17.3
`Trace
`0.3
`0.3
`0.2
`2.4
`0.4
`0.4
`11.1
`0.2
`
`1.1
`0.2
`
`32.0
`
`23.1
`44.?
`0.2
`
`Neutral
`lipid
`0.3
`0. 1
`14.3
`0.6
`1.3
`0.3
`13.4
`4.5
`2.3
`0.2
`1.6
`0.4
`1.0
`16.9
`4.2
`2.3
`3.7
`0.6
`0.1
`0.3
`0.2
`0.2
`0.7
`0.5
`12.0
`Trace
`0.7
`0.3
`0.3
`3.2
`0.4
`0.4
`5.5
`Trace
`
`1.3
`0.4
`
`37.7
`
`24.9
`31'.0
`0.4
`
`Complex
`lipid
`Trace
`—
`2.5
`0.1
`0.7
`0.1
`24.6
`3.4
`0-6
`Trace
`0.9
`0.3
`0.6
`10.6
`2.9
`1.6
`1.6
`0.2
`0.1
`0.2
`0,1
`0.3
`1.1
`0.5
`21.0
`Tract
`0.5
`0.6
`0.3
`5.6
`0.5
`0.5
`12.5
`0.2
`
`4.3
`1.0
`
`29.2
`
`16.5
`53.3
`1.0
`
`‘A: raw krill was frozen; B: bailed krill was frozen.
`
`0000011
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`0000011
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`
`U'['lLl7.ATl0N OF ANTARCTIC KHILL
`
`129
`
`Cholesterol is present in the tissue of krill at 62.1-71.6 mg per. 100 g, with lit«
`tle variafon among samples (16) as shown in Table 4. Generally, cholesterol con-
`tent of shellfish is two to four times that of fish. Krill is no exception. Although
`people may worry that eating Antarctic krill could cause an accumulation of
`cholesterol in blood, it is kr own that fat of fish and shellfish containing high
`amounts of unsaturated fatt
`y acids has the opposite effect. In fact, no increase
`in cholesterol either in blood or liver was found in rats which had been fed krill
`
`lipids for 3 weeks (27) or 2
`Krill contains r
`uch vitami
`tained in the who
`
`months (15).
`n A and other vitamins. Most of the vitamin A con-
`
`rril" is astaxanthin, which is highly concentrated in
`e body of
`the eyes. Vitamin A is reported ir widely ranging concentrations from 19? to 1,446
`IU per 100 g ofraw or boiled krill (I4). Yanase (28) measured vitamins in frozen
`whole krill. The results are surr rnarizcd in Table 6.
`
`
`
`
`
`Downloadedby[JohnJones]at18:5312September2011
`
`Inorganic substances in krill are listed in Table 7', with P as the highest in con-
`centration. followed by Na and K (29). It is reported that krill (Euphausfa .5‘!-fw
`perbifl and Meganyuctiphrmes norvgica) contain fluoride in amounts several
`times higher than other crustaceans, mainly in the exoskeleton, carapace, and
`cephalothorax, but relatively little in the muscle (30).
`
`Amount and Characteristics of Protein
`
`After separating Kl uscle {tail rr cat in Table 8), hcpatopancreas, stomach, heart,
`inner skin, and mi er remainders including carapaces of living krill sarr pics, the
`
`Table 6. Vitamin and Vitamin~Related Compounds in Krill
`
`Substances
`Vitamin A
`
`Astaxanthin
`Riboflavin
`Vitamin B.
`Ca-panthothcnatc
`Niacin
`Folic acid
`Biotin
`Vitamin Hi2
`
`Amounts in frozen whole body“
`(wet weight basis}
`330 IU/100 g
`
`3.12 mg/I00 g
`LS8 -y/g
`Ll 1-fg
`I5 -Hg
`T0 -yfg
`60 7/100 g
`10 ‘y/I00 3;
`16 7.3100 g
`
`“Determination methods: Carr-Price reaction method for vitamin A,
`spectrophotometrical method for astaxanthln with oil solved in ethyl-
`ethcr; lunillavin fluorescence method for riboflavin; microbiological
`assay for other vitamin B with hot water extract of material which is
`digested with papain.
`
`0000012
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`AKER877|TC00118592
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`0000012
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`

`
`
`
`
`
`Dowaiazzaziggby[J(::}'11'1,5t:3E:%fS§af25:5313S&p§i:‘m§3€?29::
`
`130
`
`§3U'I;l,FKi AMI) i'3M5I!A1'A
`
`Tank‘ 7.
`
`Inorganic Substances in Ksifi
`
`MineraifMMMMMMMMMMMMMMMMMMMMMMMMMMM
`r*_'M W
`Cs:
`
`WMM
`
`100 g £§_I“EqMi:rilI
`1;.§§t1-mm
`2‘72~7»63?
`
`Na.
`K
`
`Mg
`Fe
`
`l,2.‘.r3~ 1 £29‘
`1,076-3,4430
`
`94
`22.4
`
`Tame 8. Wcighl, Prmeia Cazmtezm, and pH af Various Tisgucs :11’ Faresh Krm é
`Pmtcin“ {gfbm g :3.‘ krifil}
`pH”
`’““_""‘"‘“"""""'_‘
`“""‘ ‘“““““““““““"0”
`A5
`After H
`
`Weight par
`Wham: hardy
`
`Tait mama
`
`H:pato§:an.tcrmx
`Smmach and hear!
`Inner xkin
`
`(T.‘aarzxa.'.=sm:.* mm! retrrlainimg
`mm
`
`26.4
`
`7.8
`3.0
`10.8
`
`40.7
`
`4.! (15.4?
`
`1.6 (45,103
`
`$8 (10.3)
`8.3 (3.8)
`(3.7 (6.5)
`
`40119.3}
`
`0.3 {I03}
`{L3 {$3}
`0.? {tS.5)
`
`20% ($3.9)
`
`Scparmian lass
`
`~——~
`
`-—
`
`6.2
`9.9
`100.0
`Tittal
`—-»~ «sum.
`"W’mc:-mium: prcmein wen asxmmned with phmphaz: mmu {pH 7.3, I
`”x'sH was mcmturfid using hnmz;«g;e:*nmc £111" each !i.E:9i11€ with dislmed wanna
`“Tm: vzglszm Em [:?n:if(:t1ilIc*M:§ flsxxw em»: carnm! mi of xwamn in am 3;
`:1!‘ cash. «issue.
`
`‘M
`
`6.’?
`7.2
`'?..‘i
`
`805
`
`-—-—
`
`—
`
`M
`
`6.5
`7.0
`‘L2
`
`8.8
`
`:_
`
`— _
`
`ram: £3? weight and cement of prmein afeach tissue WES measured. Tm results
`an: simwn in Tame 8 {B1}. The pmzicm of muscle: is 26.7070,‘ simiiar m results
`himmo :::mbii5hr::d (9). The portimn me“ pmmin in ma muscle ia: about MW: of ad!
`p0rou:i:: Em krill. ‘mus ii ahmws that rmzsm mzan 50% mi‘ pvrmeixm in krill is ms! dur»
`ing the mmuval mt” campmtea and inmrnm mgams.
`Abiifilt 8{l‘3'£"o of the nitmgen cumpounds in krifl muscla is proteimnitrogen,
`and myofibrillar protein makes; up 6&1-?U0% of the tote} pmtein, G1’ on {he average
`9.4 g in E00 g of muscie, similarm fish meat (31). 0:: the other hand. the par»
`timn mf mraoplaamaia protein is high in tissuas mher than mmaclcz. The: mymfibri]~
`lar garmcfin is rr1:»:fin1y czwmpmed at” mgmséin, actin. and pamamyosin, tmmgy similar
`tr) that nrf mumle mf Other invertebrams (’31»33). Tabie 9 shims prmmim mmpusi»
`ticm in min meat.
`
`0000013
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`AKER877|TC00118593
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`0000013
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`

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`
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`
`Downloadedby[JohnJones]at18:5312September20]]
`
`UTILlZA"l0l"i OF AI‘-i'l"ARC'l'IC KRILL
`
`131
`
`Table 9. Various Protein Contents in Krill Meat
`
`Sample
`l
`
`2
`
`Nonprotein-N
`(mg I‘-lfg}
`4.72
`
`4.53
`
`Protein-N (mg Hfg)
`
`Sp.“
`2.30
`1114.2)"
`
`Mi.“
`11.32
`00.0}
`12.6
`
`Alkalirsoluhle
`2.23
`{14.l}
`2.20
`
`(84.0)
`“SP4 sarcoplasrnic protein Sup I + Sup 2 in Fig. 5~fi; Mi"; rnyofihrillar protein.
`“Values in parentheses Show each protcinnbl (“Fol in total protc-in—N.
`
`Stroma
`0.25
`(1.6)
`0.22
`
`(1.5)
`
`Table 10. Amounts of Water-Soluble and Salt-Soluble Proteins
`
`Protein (Wu)
`
`Part of krill body
`Whole krill
`ail
`teat
`
`Moisture {Wu}
`SL6
`8-1.?
`
`Waterrsolublc‘
`5.] 3: (lat
`5.0 2!:
`(1.4
`
`Salt-soluble”
`3.? 1 0.4
`9.4 :t 0.8
`
`"Watcr—soluble protein was extracted by phosphate buffer solution (!5.S mM Na;Hl’O. +
`3.38 rnM KHZPOOI pH 7.5, l = 0.05};sa|t—solubie protein. by 0.45 MKCI phosphate bullet
`solution {pH ?.3,,
`= 0.5}.
`
`Previous data co cerning protein of Antarctic krill were obtained by :easur-
`ing frozen krill or processed krill brought back iron the Antarctic Ocean. Shibata
`(31) measured protein solubility of krill i
`edia eiy after each catch. "able 10
`shows the amounts of water-soluble and sa t-soluble proteins.
`Because the body of the krill is so small, its
`uscle is located close o other
`issues (mainly digestive organs} containing protease, and muscle protein is easily
`affected by protease. It is, therefore, necessary o wash the muscle wi h large
`amounts of water immediately after each catch to -prevent other t'ssues from her
`'ng mixed with the muscle.
`Iolecu-
`uscle of krill aggregates and increases its
`Myolibrillar protein in the
`ar size with tin‘ c after dea h. "he agg egation is
`uch faster tha in the case of
`o her fishes (3 }. Aggregation of nyofibrillar protei
`a d enlargemen of its
`olecular size also occur dur‘ng freezing storage (31).
`Denaturatior speed of yofbrillar protein of kr'll compared to those ofother
`ishes is shown ‘n Figure 6 (33). In this figure. the te pera ure stability of the
`Ca-A'"Pase of
`yofibrillar protein in several species of '1sh is show“: by an oblique
`jne."he‘1eso thefightfideindkamthecondhmnsatwhchthe yoibfiflar
`prote'n denatures faster than the lines on the left side. "he myofibrillar protein
`of krill is
`ore unstab e than those of rat-tail (Nematormrtrs pectorafs) or horse-
`1air crab (A..FiHIflC€l2‘S isenbackir), which have been considered very mstable by
`experfinentslutherus
`ade.
`
`0000014
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`AKER877|TC00118594
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`0000014
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`

`
`I32
`
`SUZU Kl AND SHIIIATA
`
`(‘C l
`
`50
`
`40
`
`30
`
`20
`
`10
`
`E}
`
`\
`
`1
`
`1
`
`A \
`1.
`B
`1
`
`__
`
`...
`
`.A
`
`\
`
`c.
`
`3
`
`—
`
`100:-
`
`"
`
`M
`
`10
`
`T L
`
`1
`
`"E:
`
`3;;
`
`L"?
`
`0.1
`
`‘
`
`1
`I 037-,
`
`‘
`
`Inactivation rate constants of myofibfllar Ca—ATPa.se of krill and other fishes. A—-A. .
`Figure 6.
`krill; A, rat—taiI; B, Alaska polinelc; C, sardine; 1), Whale.
`
`Adding sorbitol or sodium glutamate as antidenaturant is effective in stabiliz-
`ing such unstable proteins of krili (33). The stability of krili protein can be in~
`creased about 4 times by addirg L0 M; 16 times with 2.0 M; and about 94 times
`with 3.0 M sorbitol, respectively.
`
`Components of Taste and Smell
`
`Krill has good taste with so e sweetness similar to that of shrimp. The taste gen-
`erally exists in the extract obtained after extracting the tissue with hot water to
`
`0000015
`
`AKER877|TC00118595
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`
`
`
`
`flownioadedby[JohnJones]at18:5312September2011
`
`0000015
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`

`
`UTILIZATION OF Al‘-lTARC'l‘IC KRILL
`
`133
`
`. The extract i
`eludes free a ni o acids. peptides, orga ic
`etc. It is considered that no
`protein ni*rogen compot. nds,
`acids, in the ex
`tract play an important
`role in evoking he
`ion of "ree arr
`no acids of prawns {Fe
`noeus japonicus) is
`taste. The eompos'
`on to tha
`1uscle is rich i
`sown in comparis
`elv-
`ofkrili in Table 1] (34). Krill r
`ine, alanine, pro!’
`s as
`e, argi nine, lysine, and taurine. Moreover, it contai
`‘Iuch as 106 r1g%
`of betai c. This is somewhat similar to shrimp and irr plies
`hat other compou
`nds contribute to the taste of krill. However, the amot nt of
`he taste components such as glycine and betaine of krill is less the
`that of prawn,
`'1at is, more than 1
`t
`000 r1g% of glycine and 539 mgilb of betaine (35). It is known
`that glycine decreases during freezing storage. Accordingly, the low va ue of
`these compounds i’" kn I may be because the measurement was made on frozen
`samples. The specific smell of frozen krill was found to be caused by dimethyl-
`krill
`sulfide (DMS) and a volatile ami
`e. Tokunaga (36) rcported that froze
`contains 50-EH00 ng/g of DMS, r
`‘|(}l'C in cephalothorax than in telson. DMS
`evokes a flavor specific to crustaceans when its concentration is below about
`
`remove the prolei
`acids, nucleotides,
`such as free amino
`
`s c
`
`1''
`
`Table I1. Free Amino Acids in Krill and Prawn
`Meats
`
`Amino acid
`
`Alanine
`
`Glycine
`Valiuc
`Lcucine
`
`lsolcucinc
`Proline
`
`Phenylalanine
`
`"yrosine
`Scrinc
`hrconine
`
`Cystine
`Methionine
`
`Arginine
`Histidinc
`
`Lysine
`Aspartic acid
`fiiutamic acid
`Tauririe
`
`"80% ethanol extract.
`
`FAA content {mg/I00 :5)
`Krill
`Prawn”
`
`106
`
`llfi
`62.’!
`85.6
`
`43.4
`2]?
`
`53.3
`
`47 .6
`42.‘!
`53.6
`
`0
`33.9
`
`266
`16.5
`
`M5
`52.!)
`35.|
`206
`
`S3
`
`i250
`I9
`17
`
`1 I
`230
`
`'3’
`
`I
`I08
`15
`
`—
`1]
`
`686
`7
`
`15
`race
`65
`—
`
`0000016
`
`AKER877|TC00118596
`
`1 t
`
`
`
`
`
`Downloadedby[JohnJones]at18:5312September2011
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`0000016
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`

`
`I311
`
`WJZUKI AN}? MIWATA
`
`I00 ngfg. The amma becomes unpleasant when it exceeds the concentration
`all 3 gxgfg, and it brecomex uffensive when it exceeds several micrmgramsl/gram.
`EMS is: trmymaxlcally ‘lrrvrnled Eran: climelhyl~l3~pmpim:ethlne in phytaglanklcmns
`whic:;h smvéz as [end for l-trill. Szciimggsx M W 30 “(.3 is clasimble m pmw-ml l‘m'“ma-
`tion of IDMS;
`Zlllff is lrmzfficiem. Tlw offensive mim till krill is mzzuzmcl by the
`large amcsums of irlmeiihylazninc and imbutylmnlne in krlll (3%. Trimeihylarnine
`comes fmm trimelhylamlneoxide.
`Many wnlpounds aye formed when hill is bmllatl. Same compounds, such
`as pyridine and llllaltlllflfl, pmduce a plmsant flavor, whllc: aldelaydes cause of—«
`fctmziva <IM1llI}fSi. Tlm diwgrtteablfi Cvclm sléfififilllfi to krill is formed by a mixture of
`thma wmmzzunds. Krill mmains large ammmts cal" sulfumontainlng mmgmunds.
`Some at" these cmmpmmszls haw: low thrmihulds and thus mntrlbum in lime smell
`sptcific Em krill (3%).
`The muscle with the campaces and internal organs. removed does mm emit any
`uffmsllwr odor during sturage, tiwrzn when hemcd.
`
`NUTRXTIVE VALUE ll)? KRILL
`
`Because pmtein is {he chief compnnem of krill, it is impmtanl to rcssicw its nu-
`trillvt‘: valmr. Ntllritlvcz aevrmluation it»: by an amino acid share of the iym and quan»
`mg of eamntial amine: mills ctcrnmizxerrl our by the ram of weiglxt gain ml mus lead
`dlllercmt proteins.
`Aminu acids of lcrill gruvain haw: bean mudlszd well (313-41), and there is a re-
`semblance to those cf cammon cruszacwns such as prawn (Penauesjaplonicus)
`or shiba shrimp (llfezupemauesjaynerzj. Examples M recent amines acid analyses
`ml‘ ll-zrill pmleln are slmwn in Table: 2:2, Sulfur-mntalning amirm acids ware fcmnd
`ta ha law cvasrtparmtl m ulmsc: in W“h(3l%-€;‘gg prmein. On the Clrthm“ hamsl, lyginss
`and tlvrwnlrm, whlczh am fmund only has smnall zzmmmm in grains, me: mlalively
`abundant; in krill. The amino acid scare {method recommended by a FAEZL/WHO
`commi§£e¢:} for krill pmmin ranges fmm 85 to 100 (42; The discrepancy of re-
`sults abiajlncd by different scientists is. probably bscause of use of different math»
`ads of analyscm. The mnimfl acid acmeza all milk anal shim shrimp am 93 (41) and
`"ll (41), respectiwztly, wl1hsu|fur~cwnmmlng amirmr zacziilzoz being the prlmmjy limit»
`ing amlsm acids in beam cases. ‘Thmse rmulla mow tlml the krill prmcln is very
`similar to that of casein in milk.
`
`Fmimal experiments give more accurate estimates of protein wmlu: than the
`amlnc: acid score: lmcaus-3 the latter dam; not include digestion and abscrrptiun
`arapecta.
`llnllad lcrill, zxftszr belng frozen and dried, were: fwd 1:3 rats fm 4 wmlaia. Weight
`galn. pmteln efficlmfmy lllllillll), and met premix} utilimllun (NEW) vwzm wm«
`pared ta Elms: obtained with whole-egg pmtcin. Valum wlmaincd with ‘mill pm»-
`leim wexe lower than {haste obtained with whole-egg garmcin bu: were: similar to
`
`
`
`Dmxvzzligaaféeéiby[balm.l<;n1§:§§3i3,1&5}123f:§i{’1‘i‘z§?s‘3§‘2011
`
`0000017
`
`AKER877|TC00118597
`
`0000017
`
`

`
`UTILIZATION OF ANTARCTIC KRILL
`
`135
`
`Table I2. Amino Acid Compositions of Krill Protein (g of amino acid in hydrolysate from 100 g
`of protein)
`Amino acid
`Alanine
`
`Whole body
`5.46
`
`hlnscle
`5.83
`
`Muscle
`6.01
`
`TC/‘\l “precipitate
`6.2
`
`Glycine
`Valine
`
`Leuclne
`lsolcueine
`1'-‘roline
`
`Phenylalanine
`"yrosirie
`"ryptophane
`Serine
`
`Hhrconlne
`
`Cystine
`Methionine
`
`Argininc
`Histidine
`
`Lysine
`Aspartic acid
`Glutamic acid
`Glueosaminc
`Amide N
`
`Total amino acid
`
`N recovered (Wu)
`
`4.67
`5.90
`
`7.77
`5.10
`4.21
`
`6.47
`4 .06
`1.50
`4.95”
`
`4.70
`
`1.45
`3.03
`
`6.22
`2.30
`
`8.58
`12.20
`14.60
`3.45
`L37
`
`106.62
`
`9705
`
`4.58
`5.33
`
`8.47
`5.50
`3.36
`
`6.35
`4 .29
`1.60
`4.90“
`
`4.65
`
`1.14
`3.53
`
`6.83
`2.16
`
`9.50
`12.00
`15.00
`
`1.23
`
`106.07
`
`97.41
`
`4.62
`4.72
`
`8.28
`5.25
`3.35
`
`6.32
`4 .51
`1.70
`3.91“
`
`4.20
`
`1.35
`3.4-4
`
`7.03
`2.40
`
`10.20
`12.50
`15.30
`
`1.26
`
`105.14
`
`98.55
`
`“Values corrected by I0“l'a to compensate for destruction during acid hydrolysis.
`
`41.5
`5.5
`
`8.0
`5.8
`3.2
`
`4.6
`4 .9
`1.9
`4.7
`
`4.5
`
`1.0
`2.5
`
`6.8
`0.8
`
`9.8
`11.0
`12.5
`
`99.0
`
`
`
`
`
`Downloadedby[JohnJones]at18:5312September2011
`
`those of casein. Table 13 (42). The relative nrotein values of boiled kr’ 1 and
`casein were 87.4 and 77.6, res
`neciively, assu ning whole egg as 100, with krill
`somewhat higher than casein (43).
`ues of raw and boiled krill with rat-feedi
`Comnarison
`of utritive val
`hat raw kri
`'ZIIE]‘1l"flf:l"'|1S showed
`
`‘I had a lower value than the boiled. In addition,
`ritive values than boiled krill with heads
`boiled krill without heads had higher ml
`live v