1. P. O. Andersson, T. Gillbro, A. E. Asato & R. S. H. Liu, "Dual Singlet State Emission in a Series of MiniCarotenes," J. Luminescence, 51, 11-20 (1992).

2. S. Ganapathy & R. S. H. Liu, "Polyenes 30. Quantum Chain Processes in Photoisomerization of the
All-Trans, 7-Cis and 11-Cis Isomers of Retinal," J. Am. Chem. Soc., 114, 3459-3464 (1992).
3. R. S. H. Liu, E. Krogh, X.-Y. Li, D. Mead, L. U. Colmenares, J. R. Thiel, J. Ellis, D. Wong & A. E.
Asato, "Analyzing the Red-shift Characteristics of Azulenic, Naphthyl, Other Ring-fused and Retinyl
Pigment Analogs of
Bacteriorhodopsin," Photochem. Photobiol., 58, 701-705 (1993).
4. R. S. H. Liu, "Photochemistry of Polyenes Related to Vitamin A," in "CRC Handbook of Organic
Photochemistry and Photobiology," eds. W. M. Horspool & P. S. Song, CRC Press, 165-172, 1995.
5. L. U. Colmenares & R. S. H. Liu, "Fluorinated Phenylrhodopsin Analogs. Binding Selectivity,
Restricted Rotation and 19F-NMR Studies," Tetrahedron, 52, 109-118 (1996).
6. A. E. Asato, R. S. H. Liu, V. P. Rao & Y. M. Cai, "Azulene-containing Donor-acceptor Compounds as
Second-order Nonlinear Chromophores,"Tetrahedron Lett., 37, 419-422 (1996).
7. L. U. Colmenares, W. P. Niemczura, A. E. Asato & R. S. H. Liu, "A 19F NMR Study of Rhodopsin
Analogs: Use of Vinylfluororetinal Chromophores," J. Phys. Chem., 100, 9175-9180 (1996).
8. R. S. H. Liu & A. E. Asato, "Making Organic Concepts Visible," J. Chem. Ed., 74, 783-784 (1997).
9. D. Hoishen, L. U. Colmenares, J. Liu, C. J. Simmons, G. Britton & R. S. H. Liu, "Fluorinated
Analogs of the Carotenoprotein, a -Crustacyanin," Bioorg. Chem., 26, 365-374 (1998).
10. L. U. Colmenares, X-L. Zou, J. Liu, A. E. Asato, R. S. H. Liu, A. deLera & R. Alvarez, "11,12Difluororhodopsin and Related Odd-Numbered Fluororhodopsins. The Use of JF,F for Following a Cis-trans
Isomerization Process," J. Am. Chem. Soc., 121, 5803-5804 (1999).
Chayen, N.E., Gordon, E.J., Phillips, S.E.V., Saridakis, E.E.G. & Zagalsky, P.F. (1996) Crystallization and
initial X-ray analysis of beta-crustacyanin, the dimer of apoproteins A(2) and C-1, each with a bound
astaxanthin molecule. Acta Crystallographica Section D - Biological Crystallography 52:409-410.

http://www.mun.ca/seabright/caroteno.html
ScienceNet : Life Science - Biochemistry
http://www.sci-ctr.edu.sg/ScienceNet/cat_life/cat_bch06799.html
Welcome to Dr. R.S.H. Liu's Page
http://www.chem.hawaii.edu/UH_Chem/faculty/liu2.html
[ScienceNet Logo][ScienceNet Logo] Life Science Category
Biochemistry
Thu Feb 24 11:31:28 SST 2000 6799
Lisa Mak lisamak@interchange.ubc.ca
21 to 30 Student Post Graduate
Life Science
When shrimps are raw, they are greyish-green. So, how and why do they
turn
red when you cook them?
Answer : Carotenoids, both carotenes and xanthophylls, are widespread
amongst animal groups and the species within the groups. In many animals,

carotenoids are linked to proteins and the resulting carotenoproteins may

be different in colour to the free carotenoids. Whereas free carotenoids
are fat-soluble, carotenoproteins may also be soluble in water. Unlike
carotenoids, carotenoproteins often change colour with acidity or heat.
Carotenoids are widespread in crustaceans, and the blues and greens of
most crabs, prawns and lobsters are due to carotenoproteins, often in
conjunction with free carotenoids. When shrimps, lobsters and crabs are
cooked, the heat
breaks down the carotenoprotein: protein and pigment
separate and in the process the pigment is oxidised. Blue, green, black
and purplish red colours all change to the orange-red colour of astacin,
the oxidised product of astoxanthin. In this case, the free red pigment
is
brighter
than
most
of
the
carotenoprotein colours.

-----------------------------------------------------------------------(c) Copyright 1997 Virtual Science

Centre Project Team
Rntgenstrukturaufklrung, Dynamik und Kleinwinkelstreuung von
Proteinkomplexen
Dynamik von Myoglobin
(AG H. Hartmann in Zusammenarbeit mit Prof. F. Parak, TU Mnchen, bis
1996)
Von Myoglobin wurden Mbauerspektren im Temperaturbereich von 90K-255K
gemessen und mit empirischen Relaxationsmodellen nach Davidson-Cole, oleCole und Havriliak-Negami analysiert, die bei organischen Glsern seit
langer Zeit erfolgreich verwendet werden. Die bereinstimmung der
Glasmodelle mit den gemessenen Spektren ist besser als mit speziell fr
die Proteindynamik entwickelten Modellen, die auf stochastischen
Bewegungen der Atome in einem begrenzten Raum beruhen. Oberhalb einer
Temperatur von 200K ergibt sich eine sehr breite Verteilung fr die
Fluktuationszeiten der Atome. Beides kann man als Hinweise auf eine
hnlichkeit von Proteinen und organischen Glsern ansehen. Andererseits
finden wir aber eine ungewhnliche Temperaturabhngigkeit des Parameters,
der die Breite der Verteilung beschreibt. Fr Myoglobin nimmt die Breite
der Verteilungsfunktion von 190K bis 255K stetig mit der Temperatur zu,
whrend man bei einer Analogie zu organischen Glsern zumindest in einem
bestimmten Temperaturbereich einen konstanten Wert erwarten wrde.
-- Hartmann H., S. Zinser, P. Komninos, F. Parak, R.T. Schneider and G.U.
Nienhaus (1996). X-ray structure determination of a metastable state
of carbonmonoxy myoglobin after photodissociation. Proc. Natl. Acad.
Sci. USA 93:7013-7016
-- Chang I., H. Hartmann, Y. Krupyanskii, A. Zharikov and F. Parak
(1996).
Dielectric relaxation models applied to the dynamics of myoglobin as
determined by Mssbauer spectroscopy. Chemical Physics 212:221-229
Kristallisation von Proteinen und Rntgenstrukturaufklrung (H. Hartmann,
A. Bongers, R. Gebhardt, B. Lohkamp, N. Smilyanska, H. Decker;
Zusammenarbeit mit Dr. M. Wilmans, EMBL Hamburg)

Verschiedene Proteinkomplexe wie Myoglobin, Hmocyanine aus Arthropoden

und
Mollusken sowie ein Porenprotein aus Mollusken (in Kooperation mit Prof.
J.
Markl, Inst. f. Zoologie, Univ. Mainz) werden kristallisiert. Die Daten
werden hier am Institut mit unserer Drehanode und einer Anfang 1998 neu
installierten Image-Plate aufgenommen. Einzelne, wenn auch recht kleine
Kristalle des 24-meren Spinnen-Hmocyanins, der 12-meren Astacus und
Homarus Hmocyanine und des 6-meren Panulirus-Hmocyanins streuen mit
einer Auflsung von 3 . Grere Datensets werden am EMBL in Hamburg
aufgenommen.
Ein blaues Carotenoprotein aus dem Schwamm Suberites domuncula (E.
Jaenicke, H. Decker)
Blaue Carotenoproteine finden sich in vielen marinen Invertebraten. Durch
ihre Frbung tragen sie dort zum Schutz vor Rubern bei. Diese
Carotenoproteine bestehen aus einem Apoprotein und einem nicht-kovalent
gebundenem Carotenoid, welches in vielen Fllen Astaxanthin oder
Canthaxanthin ist. Durch die Bindung an das Apoprotein verndert das
Carotenoid seine Farbe von rot nach blau. Das bekannteste und am
intensivsten untersuchte blaue Carotenoprotein ist das Crustacyanin, das
dem Carapax des europischen Hummers seine charakteristische blaue Farbe
verleiht. Die thermische Denaturierung des Crustacyanins und die damit
verbundene Freisetzung des Carotenoids ist auch die Ursache, weshalb ein
Hummer beim Kochen seine Farbe von blau nach rot ndert.
Die Natur der spezifischen Carotenoid-Protein Wechselwirkung in den
blauen
Carotenoproteinen ist zum einen hinsichtlich des Mechanismus von
spektralen
Verschiebungsphnomen in Polyenen im allgemeinen und darberhinaus fr
biologische Prozesse wie die Photosynthese, dem Schutz vor schdigender
Lichtstrahlung, dem Sehen und der Wechselwirkungen von Vitamin A und
seinen
Derivaten mit Proteinen von Interesse.
Wir haben eine blaues Carotenoprotein aus dem marinen Schwamm Suberites
domuncula isoliert und spektroskopisch charakterisiert. Durch
CD-Spektroskopie im fernen UV konnte die Zugehrigkeit dieses
Carotenoproteins zu den Lipocalinen besttigt werden. Die Kristalle des
Carotenoproteins sind dunkelblau gefrbt und haben bisher eine Gre von
50x50x50m.
Rntgen- und Neutronenkleinwinkelstreuung an Proteinkomplexen
(H. Hartmann, B. Lohkamp, A. Bongers, R. Gebhardt, H. Decker; seit 1995,
Zusammenarbeit mit PD Dr. T. Nawroth, Koordinater einer offenen
Strahlgruppe, Prof. C. Brandt-Koch, beide am Institut fr Biochemie,
Universitt Mainz, und PD Dr. H. Heumann, MPI fr Biochemie Martinsried;
die Experimente werden am MPI fr Polymerchemie, Inst. f. Physikalische
Chemie der Universitt Mainz und am DESY, Hamburg, (HASYLAB: Dr.
G.Goerigk;
EMBL: Dr. G. Rapp) und am ILL Grenoble (Dr. M. May) durchgefhrt). Dieser
Forschungszweig wurde am Institut fr Molekulare Biophysik zu einem
Schwerpunkt ausgebaut. Die Daten werden am HASYLAB; DESY Hamburg aund am
ILL Grenoble gewonnen.
Die Kooperativitt respiratorischer Proteine setzt die Existenz

verschiedener Konformationen voraus, die durch besondere

Sauerstoffbindungseigenschaften gekennzeichnet sind. Die Existenz von
zwei
Konformationen, einem oxygenierten Zustand und einem desoxygenierten
Zustand, konnten wir mit Hilfe der Rntgenkleinwinkelstreuung zeigen. Die
oxygenierte Form scheint dabei weniger kompakt zu sein, als die
desoxygenierte Form. Mit der Oxygenierung geht eine Lngennderung in dem
untersuchten 24-meren Vogelspinnen-Hmocyanin von 25nm auf 27nm einher.
Verschiedene Modelle, die aus elektronenmikroskopischen Bildern und
Rntgenstrukturen abgeleitet sind, wurden an die Streukurven angepat.
Anhand der Parameter in diesen Modellen konnten wir feststellen, da die
oben erwhnte Bewegung nicht nur auf der 24-meren Ebene, sondern auch auf
der hexameren Ebene, der kleinsten in vivo vorkommenden Form von
Hmocyanin, auftritt. Wir konnten auch fr andere Hmocyanine zeigen, da
die aus den elektronenmikroskopischen Bildern abgeleiteten Dimensionen
modifiziert werden mssen, so z. B. die Abstnde zwischen den beiden
Halbmoleklen. Rntgenkleinwinkel-Streuung zeigt
Konformationsunterschiede
von Hmocyanin in Lsung, die in fixierten Formen, wie z.B. in
Rntgenstrukuturanalysen, nicht gefunden wurden.
Rntgenkleinwinkel-Streuversuche am 12-meren Hummer-Hmocyanin zeigten
jedoch, da die oxy-Form kompakter ist als die desoxy-Form. Der pH-Wert
hat
einen fast nicht nachweisbaren Einflu auf die Form dieses Hmocyanins,
ebenso wie der Effektor Laktat.
Seit kurzer Zeit untersuchen wir in Zusammenarbeit mit Prof. J. Markl
(Institut for Zoologie, Universitt Mainz) das Hmocyanin (KLH) aus der
Schlssellochschnecke Megathura crenulata. Die Aufklrung der Struktur
und
immunologischen Wirkungsweise dieses Molluskenhmocyanins ist ein
Forschungsschwerpunkt der AG Markl. Im Vergleich zu
elektronenmikroskopischen Aufnahmen konnten wir fr das didekamere KLH
eine
bereinstimmung hinsichtlich der Form finden, wobei wir etwas kleinere
Dimensionen fanden. Erste Ergebnisse zeigten auch groe Unterschiede fr
das dekamere KLH im oxy- und im desoxy-Zustand.
-- Decker H., H. Hartmann, R. Sterner, E. Schwarz and I. Pilz (1996).
Small-angle X-ray scattering reveals differences between the quaternary
structures of oxygenated and deoxygenated tarantula hemocyanin. FEBS
Letters 393:226-230
-- Lohkamp, Bernhard (Diplomarbeit, Universitt Mainz 1997).
Rntgenkleinwinkelstreuung an Hmocyaninen.
Hartmann H., B. Lohkamp and H. Decker (1997). Modelbuilding of
multimeric proteins from small-angle X-ray data with partially known Xray
structures. Eur. Biophys. J. 26:1/63
-- Lohkamp B., H. Hartmann, H. Decker, M. Rssle, H. Heumann, G. Goerigk,
A. Post, C. Koch-Brandt and T. Nawroth (1996). Structural
dynamics of the oxygen transporting metallo-protein hemocyanin.
HASYLAB
Jahresbericht 1996, Annual Report II: 151-152
-- Lohkamp B., H. Hartmann and H. Decker (1997). Characterization of
structural transitions in arthropod hemocyanins by small-angle X-ray

scattering (SAXS). Eur. Biophys. J. 26:1/79

-- Bongers A., H. Hartmann, H. Decker, T. Nawroth, A. Post and G. Goerigk
(1997). SAXS measurements of keyhole limpet hemocyanin (KLH1)
in the didecameric form. HASYLAB Jahresbericht, Annual Report I:
653-654
-- Lohkamp B., G. Rapp, H. Hartmann and H. Decker (1997). Small-angle Xray
scattering of decameric keyhole limpet hemocyanin (KLH1) in
oxygenated and deoxygenated states reveals large structural
differences.
HASYLAB Jahresbericht, Annual Report II: 187-188
-- Bongers A., H. Hartmann and H. Decker (1999). Quaternary structure and
conformational transitions of keyhole limpet hemocyanin. Interner
Experimental Report, ILL Grenoble, France
-- Bongers, Andr (Diplomarbeit, Universitt Mainz 1999). Rntgen- und
Neutronenkleinwinkel-Streuung am didekameren KLH.
-- Gebhardt R., I. Lauer, T. Nawroth, H. Decker, G. Goerigk and G.v.
Krosigk (2000). Structural changes of the oxygen transport protein
hemocyanin detected by pH dependent USAXS. HASYLAB Jahresbericht
1999,
Annual Report: 979-980
-- Hartmann H., A. Bongers and H. Decker. Small angle X-ray and neutron
scattering data reveal an oxygen dependent conformational change
of the immunogen keyhole limpet hemocyanin type 1 (KLH1). will be
submitted
zurck

Extraction Of Carotenoproteins From

Crustacean Wastes
. Extraction Of Carotenoproteins From Crustacean Wastes
http://www.mun.ca/seabright/caroteno.html
Investigators
Dr. N.F. Haard
Institute of Marine Resources
College of Agricultural and Environmental Sciences
University of California, Davis, CA
Dr. B.K. Simpson
MacDonald College
McGill University, Montreal, PQ
Description
One of the problems facing aquaculturists is that species such as salmon and trout raised on a diet of fish meal
lack the customary color of the wild fish. Carotenoid pigments can be used to correct this deficiency in
rearing salmon and trout in fish farms. The pigments in the shells and tissues of crustaceans have been found
to impart the necessary color to fish flesh. Thus, the extraction of carotenoprotein from crustacean wastes
and its addition to the rations of pen reared salmonids may be used to give the product the desirable orangered color.
Technology
Researchers at Memorial University have developed a process for extracting these desired carotenoproteins
from crustacean wastes with a recovery rate of approximately 73 percent for astaxanthin (the colorant) and up
to 90 percent of protein recovered. Extraction of carotenoprotein from crustacean offal improves the nutritive
value of the raw material by reducing the content of ash and chitin and effectively concentrating the protein.
The new method consists of treating crustacean waste with a protease enzyme (trypsin) to release the
carotenoprotein, and then separating the carotenoprotein from the solid residue. As a pretreatment step,
treating the crustacean waste with a chelating agent may be employed. Carotenoprotein products made by
this method can be used as animal food supplements.
One specific benefit of this product is that, since salmonids have a high requirement for dietary protein but
excess chitin can be undesirable, the use of the extract as a feed supplement is superior to the feed value of
shrimp and similar wastes in their unprocessed form.
Status
Canadian patent No. 1,313,835 issued Feb.23, 1993. Available for licensing.
Contact
Seabright Corporation Limited
Memorial University of Newfoundland
St. John's, Newfoundland
Canada, A1C 5S7
Tel. (709) 737-4527
Fax. (709) 737-4029
E-Mail. seabrt@morgan.ucs.mun.ca
WWW. http://www.mun.ca/seabright
http://www.astaxanthin.org/chemforms.htm
Astaxanthin has chemical features that result in the existence of several forms of
astaxanthin:
Stereoisomers. Astaxanthin has two chiral (pronounced "ky-ral"), or asymmetric, centers. These are the
carbons numbered 3 and 3' (pronounced "three prime") on the two rings in the structure. One can think of
chiral asymmetry as analogous to "handedness". A left hand and a right hand are mirror images of each
other--they are similar but not identical, and are not superimposable. Similarly, a chiral center can
exist in either of two configurations; the same atoms are bonded to the chiral center, but the three-dimensional
arrangements are different and not superimposable. Chemists identify chiral centers as being either R or S

(from rectus or sinister, Latin for "right" or "left"). The two chiral centers in astaxanthin, carbons 3 and 3', can
each exist either in the R or the S form, and thus there are a total of three stereoisomers:
3S,3'S, 3R, 3'S, or 3R,3'R. The 3S,3'S and 3R,3'R stereoisomers are mirror images of each other and are
termed "enantiomers". Each enantiomer has the opposite optical activity of the other, i.e., a solution of a pure
enantiomer will rotate plane-polarized light in a direction opposite to that observed for the other enantiomer.
The 3R,3'S form is sometimes termed "meso" and is optically inactive because there is a plane of symmetry
through the center of the molecule.
How is synthetic astaxanthin different from natural astaxanthin?
Synthetic astaxanthin is produced as the free (unesterified) xanthophyll and as a
1:2:1 mixture of the three stereoisomers: 3S,3'S, 3R,3'S, and 3R,3'R. The industrial producers of synthetic
astaxanthin are Hoffmann-La Roche AG and BASF AG.
Are there different forms of natural astaxanthin?
In its natural state, astaxanthin is usually associated with other molecules (Bernhard,
1990). It is often complexed with proteins, producing an array of colors in different
organisms.
For example, it is the chromophore in the blue, green, and yellow pigments of lobsters. In other cases,
astaxanthin may simply be dissolved in the lipid fraction of complex molecules such as egg lipoproteins, or it
may actually be bound chemically to molecules such as fatty acids to form esters. Reddening of some snow
algae (Bidigare et al. 1993) and Haematococcus is the result of such esters
accumulating in
cytoplasmic lipid droplets. Less often, because it is not as stable, astaxanthin occurs in cells as a free,
unbound molecule.
Whether free or complexed, the atoms comprising an astaxanthin molecule can be
oriented in different ways, producing different isomers. The most common geometric
configuration in both synthetic and natural astaxanthin is the most thermodynamically stable all-E (all-trans)
isomer. Astaxanthin from natural sources tends to occur predominantly as either the 3S,3'S or 3R,3'R form,
while the meso (3R,3'S) isomer is the most abundant in synthetic astaxanthin (Bernhard 1990).
Why do we think natural astaxanthin may act differently from synthetic
astaxanthin?
All-E isomers are the major geometric isomers in both synthetic and natural
astaxanthin (Turujman et al. 1997). However, synthetic astaxanthin is produced as free (unesterified)
astaxanthin in a mixture of stereoisomers: the stereoisomers (3R,3'R), (3R,3'S) and (3S,3'S) occur in a ratio of
1:2:1. Natural astaxanthin, on the other hand, is usually esterified and predominantly of (3S,3'S) configuration
or, less frequently, mainly (3R,3'R) (Bernhard 1990). In Haematococcus pluvialis, astaxanthin occurs as the
3S,3'S stereoisomer and primarily as monoesters (>90%), with diesters comprising ~8% and the free molecule
~1% (Renstrm et al. 1981). It tends to produce higher pigmentation in rainbow trout compared to synthetic
astaxanthin provided at the same dietary concentration (Bowen et al., 1999).
Bernhard, K. Synthetic astaxanthin. The route of a carotenoid from research to commercialization. In:
"Carotenoids: Chemistry and Biology," N. I. Krinsky et al. (editors), Plenum Press, New York, 1990, pp. 337363.
Bidigare, R.R., Ondrusek, M.E., Kennicutt, M.C., II, Iturriaga, R., Harvey, H.R., Hoham, R.W., and Macko,
S.A. (1993) Evidence for a photoprotective function for secondary carotenoids of snow algae. J. Phycol., 29:
427-434.
Foss, P., Renstrm, B., and Liaaen-Jensen, S. (1987) Natural occurrence of enantiomeric and meso
astaxanthin. 7. Crustaceans including zooplankton. Comp. Biochem. and Physiol. B, 86B: 313-314.

Renstrm, B. and Liaaen-Jensen, S. (1981) Fatty acid composition of some esterified carotenols. Comp.
Biochem. Physiol. B, 69:625-627.
sterlie, M., Bjerkeng, B., and Liaaen-Jensen, S. (1999a) On bioavailability and deposition of bent Z-isomers
of astaxanthin. Proceedings of the First International Congress on Pigments in Food Technology, Sevilla,
Spain, 24-26 March 1999, pp.157-161.
sterlie, M., Bjerkeng, B., and Liaaen-Jensen, S. (1999b) Blood appearance and distribution of astaxanthin
E/Z siomers among plasma lipoproteins in humans administered a single meal with astaxanthin. Abstract 2A13. Abstracts of the Twelfth International Carotenoid Symposium, Cairns, Australia, 18-23 July 1999, p. 72.
sterlie, M., Bjerkeng, B., and Liaaen-Jensen, S. (1999c) Accumulation of astaxanthin all-E, 9Z and 13Z
geometrical isomers and 3 and 3' RS optical isomers in rainbow trout (Oncorhynchus mykiss) is selective. J.
Nutr., 129:391-398.
How do carotenoids prevent oxidation?
All carotenoids share a structural feature termed "polyunsaturation", that is to say, they have several
"unsaturated" or "double" bonds (double bonds between two adjacent carbon atoms). This is in contrast to
"saturated" or "single" bonds. When double bonds are arranged in a series alternating with single bonds, they
are termed "conjugated"--this means that the electrons that make up the double bonds in the linear chain are
"delocalized", or shared evenly over the whole chain. This makes the whole chain relatively electron-rich.
This conjugated double bond structure is responsible for the characteristic yellow, orange, or red colors
typical of carotenoids. The difference in colors depends primarily on how many double bonds are conjugated,
or in other words, over how long a chain the electrons can be delocalized. Conjugated double bonds are very
chemically stable, yet are capable of specific chemical reactions that require their electron-rich yet stable
structure. For example, if a carotenoid loses one electron and becomes a carotenoid cation (positively charged
ion), the resulting charge of +1 is distributed over the electron-rich chain, a much more stable situation
than if the charge were limited to a single location on the compound.
In mammalian and human cells, carotenoids protect from oxidative damage by two general mechanisms:
quenching of singlet oxygen and dissipating the energy as heat, and scavenging of radicals to prevent or
terminate chain reactions. In photosynthetic cells (as in plants and algae), and perhaps in cells exposed to
high light levels, there are additional protective mechanisms, including: reacting with other energetically
excited molecules (chlorophyll in particular) and preventing the formation of singlet oxygen, and dissipation
of excess excitation energy through the xanthophyll cycle.
A more detailed discussion of the radical-scavenging and singlet oxygen-quenching mechanisms of
carotenoids can be found in the mechanisms of carotenoid Antioxidant Behavior
How does astaxanthin compare as an antioxidant with other carotenoids?
Several studies have compared astaxanthin's antioxidant activity with that of other carotenoids. It should be
kept in mind when reviewing these studies that measurement of antioxidant activity is highly dependent on
the experimental system used, and one should be cautious in comparing results of separate studies, or of
extending the conclusions of a given study beyond its experimental limits.
As is the case with other carotenoids, astaxanthin is a potent quencher of singlet oxygen. One comprehensive
study found astaxanthin to be twice as effective as beta-carotene (and about 80 times more effective than
vitamin E) in quenching singlet oxygen in chemical solution (Di Mascio et al . 1991); lycopene was found to
be about a third more effective than astaxanthin. Similar results were found by researchers working with an
in vitro system of human blood cells treated with different carotenoids and then exposed to singlet oxygen;
again, lycopene was found to be more effective than astaxanthin, which in turn was more effective than betacarotene (Tinkler et al. 1994).

A second major antioxidant role of carotenoids is in the scavenging of free radicals. An elegant study of
carotenoid-radical reactions in chemical solution clearly demonstrated that reactivity rates depend not only on
the carotenoid but also on the nature of the radical (Mortensen et al. 1997). In one study, astaxanthin was
approximately as effective as canthaxanthin (a xanthophyll structurally similar to astaxanthin), and about
50% more effective than beta-carotene and zeaxanthin, in preventing fatty acid peroxidation in chemical
solution (Terao 1989). In a membrane model, astaxanthin was found to be more effective at scavenging
peroxyl radicals than was beta-carotene (Palozza and Krinsky 1992). Another study using membrane models
found similar results, with astaxanthin better at delaying lipid peroxidation than zeaxanthin, canthaxanthin, or
beta-carotene (Lim et al. 1992). A tissue culture model demonstrated that astaxanthin was superior to betacarotene or vitamin E in protecting the cells from herbicide-induced oxidative stress (Lawlor and O'Brien
1995).
References:
Di Mascio, P., Murphy, M. E., and Sies, H. (1991) Antioxidant defense systems: the role of carotenoids,
tocopherols, and thiols. Am. J. Clin. Nutr., 53:194S-200S.
Tinkler, J. H., Bhm, F., Schalch, W., and Truscott, T. G. (1994) Dietary carotenoids protect human cells from
damage. J. Photochem. Photobiol. B, 26:283-285.
Mortensen, A., Skibsted, L. H., Sampson, J., Rice-Evans, C., and Everett, S. A. (1997) Comparative
mechanisms and rates of free radical scavenging by carotenoid antioxidants. FEBS Letters, 418:91-97.
Terao, J. (1989) Antioxidant activity of beta-carotene-related carotenoids in solution. Lipids, 24:659-661.
Palozza, P. and Krinsky, N. I. (1992) Astaxanthin and canthaxanthin are potent antioxidants in a membrane
model. Arch. Biochem. Biophys., 297:291-295.
Lim, B. P., Nagao, A., Terao, J., Tanaka, K., Suzuki, T., and Takama, K. (1992) Antioxidant activity of
xanthophylls on peroxyl radical-mediated phospholipid peroxidation. Biochim. Biophys. Acta, 1126:178-184.
Lawlor, S. M. and O'Brien, N. M. (1995) Astaxanthin: antioxidant effects in chicken embryo fibroblasts. Nutr.
Res., 15:1695-1704.

In what diseases has oxidation been implicated?

Many human diseases and degenerative processes have been linked in some way to the action of free radicals.
Free radicals are not necessarily the only cause for these conditions, but may well make the human body
more susceptible to other disease-initiating factors, may enhance the progression of diseases, and may inhibit
the body's own defenses and repair processes. The following conditions involving multiple organs have all
been linked to free radicals (Cross et al. 1987):
Cancer Aging (including immune deficiency with aging and premature aging disorders)
Radiation injury
Alcohol damage
Ischemia-reperfusion injuries
Inflammatory-immune injuries (including vasculitis from drugs and hepatitis B virus, idiopathic and
membranous glomerulonephritis, and autoimmune diseases)
Reactions induced by drugs and toxins
Iron overload (including idiopathic hemochromatosis, dietary iron overload, thalassemia and other chronic
anemias)
Amyloid diseases

In addition, a number of single-organ conditions have been related to free radicals (Cross et al. 1987):
Affecting the brain--senile dementia, neurotoxin reactions, hyperbaric oxygen effects, Parkinson's disease,
cerebral trauma, hypertensive cerebrovascular injury, allergic encephalomyelitis and other demyelinating
diseases, neuronal ceroid lipofuscinoses, ataxia-telangiectasia syndrome, potentiation of traumatic injury,
aluminum overload
Affecting erythrocytes (red blood cells)--lead poisoning, protoporphyrin photo-oxidation, malaria, sickle-cell
anemia, favism, Fanconi anemia Affecting the lungs--emphysema, hyperoxia, cigarette-smoke effects,
oxidant pollutant effects, acute respiratory distress syndrome, bronchopulmonary dysplasia, mineral dust
pneumoconiosis, bleomycin toxicity, paraquat toxicity Affecting the heart and cardiovascular system-atherosclerosis, stroke, doxorubicin toxicity, peripheral circulation problems, Keshan disease (selenium
deficiency), alcohol cardiomyopathy Affecting the kidney--renal graft rejection, nephritic antiglomerular
basement membrane disease, heavy metal nephrotoxicity, aminoglycoside nephrotoxicity
Affecting joints--rheumatoid arthritis Affecting the gastrointestinal tract and liver--endotoxin liver injury,
carbon tetrachloride liver injury, diabetogenic action of alloxan, free fatty acid-induced pancreatitis,
abetalipoproteinemia, nonsteroidal anti-inflammatory drug-induced lesions Affecting the skin--sunburn and
solar radiation injury, thermal injury, porphyria, contact dermatitis, Bloom syndrome, effects of photosensitive
dyes Affecting the eyes--age-related macular degeneration, ocular hemorrhage, degenerative retinal damage,
cataractogenesis, retinopathy of prematurity, photic retinopathy
It is quite clear that human health depends to a large extent on the body's ability to control free radicals and
thus reduce oxidative damage to tissues, cells, and DNA. To that end, antioxidants play an essential role in
disease prevention, in longevity, and in overall well-being.
References:
Cross, C. E., B. Halliwell, E.T. Borish, W.A. Pryor, B.N. Ames, R.L. Saul, J.M. McCord, and D.
Harman. (1987) Oxygen radicals and human disease. Ann. Intern. Med., 107:526-545.
What is the evidence that dietary astaxanthin acts as a potent antioxidant?
Astaxanthin, like vitamin E, is a lipophilic (fat-soluble) antioxidant, and thus might be expected to exert its
antioxidant properties in lipid-rich cell membranes and tissues. It was shown in two published studies that, in
rats deprived of vitamin E, the resistance of lipids (fats) to oxidation was largely restored by feeding the
animals astaxanthin (Kurashige et al. 1990; Miki 1991). In the first study (Kurashige et al. 1990), rats were
fed a vitamin E-deficient diet, with or without astaxanthin supplementation at 1 mg per 100 mg feed, for two
to four months; control rats received a vitamin E-sufficient diet. Mitochondria were isolated from liver, and
erythrocyte ghosts prepared from blood samples. Membrane preparations (intact mitochondria or erythrocyte
ghosts) were subjected to oxidation by superoxide generated in situ by an iron-catalyzed xanthine oxidase
system. An index of lipid peroxidation, the formation of thiobarbituric acid-reactive (TBA-reactive)
substances, was measured colorimetrically. Mitochondria from vitamin E-deficient rats were preincubated
with various concentrations of either astaxanthin or vitamin E, and then exposed to superoxide. The percent
inhibition of TBA-reactant formation (as compared to controls with no added antioxidant) was measured. At
all concentrations tested, astaxanthin inhibited the formation of TBA-reactants more effectively than did
vitamin E, and the IC50 concentration was approximately 3 orders of magnitude lower for astaxanthin than
for vitamin E. In this system, astaxanthin was a more
effective antioxidant than was vitamin E. TBAreactive metabolite levels were compared between erythrocyte ghost preparations from the three treatment
groups. The amount of TBA-reactants was about 15-fold greater in the erythrocyte ghosts from vitamin Edeficient rats than in those from the control group. In erythrocyte ghosts from the astaxanthin-supplemented,
vitamin E-deficient rats, the level of TBA-reactants was elevated only about 6-fold over that of the controls, i.
e., 2.5-fold less than in the vitamin E-deficient, non-astaxanthin supplemented animals. This indicates that
dietary administration of astaxanthin, in the absence of vitamin E, partially restored the in vitro oxidation
resistance of erythrocyte membrane ghosts to the levels found in vitamin E-sufficient rats.

In the second study (Miki 1991), rats were fed a vitamin E-deficient diet, with or without astaxanthin
supplementation at 1 mg per 100 mg feed, for four weeks; control rats received a vitamin E-sufficient diet.
The susceptibility to oxidation of erythrocyte ghosts was assayed by a similar xanthine oxidase-generated
superoxide system and measurement of TBA-reactants. Lipid peroxidation was high in the vitamin Edeficient rats and negligible in control animals. Rats that received the astaxanthin supplementation had
peroxidation levels about half that of the vitamin E-deficient rats, again indicating that dietary astaxanthin
restored much though not all of the vitamin E-dependent oxidative resistance of the erythrocyte membranes.
References:
Kurashige, M., Okimasu, E., Inoue, M., and Utsumi, K. (1990) Inhibition of oxidative injury of biological
membranes by astaxanthin. Physiol. Chem. Pys. & Med. NMR, 22:27-38.
Miki, W. (1991) Biological functions and activities of animal carotenoids. Pure Appl. Chem., 63(1):141-146.