Veterinarni Medicina, 58, 2013 (4): 187205 Review Article

187
Carrageenan: a review
J.Nvcs,L.Bv:osiov
FacultyofMedicineandDentistry,PalackyUniversity,Olomouc,CzechRepublic
ABSTRACT:Carrageenanisanaturalcarbohydrate(polysaccharide)obtainedfromedibleredseaweeds.The
nameCarrageenanisderivedfromtheChondruscrispusspeciesofseaweedknownasCarrageenMossorIrish
MossinEngland,andCarraigininIreland.CarraiginhasbeenusedinIrelandsince400ADasagelatinandasa
homeremedytocurecoughsandcolds.ItgrowsalongthecoastsofNorthAmericaandEurope.Carrageenans
areusedinavarietyofcommercialapplicationsasgelling,thickening,andstabilisingagents,especiallyinfood
productsandsauces.Asidefromthesefunctions,carrageenansareusedinexperimentalmedicine,pharmaceutical
formulations,cosmetics,andindustrialapplications.
Keywords:carrageenan,pharmacokinetics,toxicity,biologicalactivity
List of abbreviations
3,6-AG=3,6,anhydro-galactose,5-HT=5-hydroxytryptamine,6-APA=6-aminopenicillanicacid,ADI=acceptable
dailyintake,AG=aminoguanidine,AIDS=acquiredimmunedeficiencysyndrome,AT-III=anti-thrombin-III,
bw=bodyweight,COX-2=cyclooxygenase-2,eNOS=endothelialnitricoxidesynthase,FAO=foodandagriculture
organization,HC-II=heparinco-factorII,HIV=humanimmunodeficiencyvirus,HPV =humanpapillomavirus,
HSV=herpessimplexvirus,IFAC=internationalfoodadditivescouncil,iNOS=induciblenitricoxidesynthase,
JECFA=JointFAOiWHOExpertCommitteeonFoodAdditives,L-NMMA=NG-monomethyl-i-arginine,nNOS
=neuronalnitricoxidesynthase,NO=nitricoxide,NOS =nitricoxidesynthase,NSAIDs =non-steroidalanti-
inflammatorydrugs, PGs=prostaglandins,SSL=sodiumstearoyllactylate,TNF-=tumournecrosisfactor-
Contents
1.Introduction
2.Chemicalandphysicalpropertiesofcarrageenan
2.1.Chemicalstructure
2.2.Propertiesofcarrageenan
3.Pharmacokineticsofcarrageenan
3.1.Absorption,distribution,metabolismand
excretion(ADME)
3.2.Degradationofcarrageenaninthegastroin-
testinaltract
4.Toxicityofcarrageenan
4.1.Acutetoxicity
4.2.Short-termstudiesoftoxicity
4.3.Long-termstudiesoftoxicityandcarcinogenicity
5.Biologicalactivityofcarrageenan
5.1.Carrageenan-inducedpawoedema
5.2.Intestinalinflammation
5.3.Anticoagulantandantithromboticactivity
5.4.Antiviralactivity
5.5.Anti-tumourandimmunomodulatoryactivity
5.6.Otherbiologicalactivities
6.Usesofcarrageenan
6.1.Industrialusesofcarrageenan
6.2.Industrialfoodapplications
6.3.Pharmaceuticalapplications
6.3.1.Tetracyclineandchlorotetracycline
production
6.3.2.Semi-syntheticantibioticproduction
6.3.3.o-asparticacidproduction
6.4.Cleaninginindustrialeffluents
6.5.Otherusesofcarrageenan
7.Conclusion
8.References
Review Article Veterinarni Medicina, 58, 2013 (4): 187205
188
1. INTRODUCTION
Carrageenan is a generic name for a family of
gel-forming and viscosif ying polysaccharides,
which are obtained by extraction from certain
speciesofredseaweeds(Table1).Carrageenanis
derived from a number of seaweeds of the class
Rhodophyceae.Thisparticulartypeofseaweedis
commonintheAtlanticOceannearBritain,Europe
andNorthAmerica.Whenusedinfoodproducts,
carrageenanhastheEUadditiveE-numberE407
orE407a.E407ahasaslightlydifferentcomposi-
tion,moreover,itcontainsaconsiderableamount
ofcellulose.Carrageenanhasnonutritionalvalue
andisusedinfoodpreparationforitsgelling,thick-
ening,andemulsifyingproperties(VandeVelde
et al. 2002) and in pharmaceutical applications
(TakamatsuandTosa1993,citedVandeVeldeet
al.2002)andexperimentalmedicinethissubstance
isoftenusedforthetestingofanti-inflammatory
agents(ZacharopoulosandPhillips1997).
2. Chemical and physical properties
ofcarrageenan
2.1. Chemical structure (Figure1)
Carrageenanisasulfatedpolygalactanwith15to
40ofester-sulfatecontentandanaveragerelative
molecularmasswellabove100kDa.Itisformedby
alternateunitsofo-galactoseand3,6-anhydro-ga-
lactose(3,6-AG)joinedby-1,3and-1,4-glycosidic
linkage.Carrageenanisclassiedintovarioustypes
suchas,,,,,allcontaining22to35sulphate
groups.Tisclassicationwasmadebasedonits
solubilityinpotassiumchloride.Teprimarydier-
enceswhichinuencethepropertiesofcarrageenan
typearethenumberandpositionofestersulfate
groupsaswellasthecontentof3.6-AG.Tesenames
donotreectdenitivechemicalstructuresbutonly
generaldierencesinthecompositionanddegree
of sulfation at specific locations in the polymer.
Higherlevelsofestersulfatemeanlowersolubil-
Table1.Characteristicsofcarrageenan(adoptedfrom:Tobacman2001)
Chemical
composition
hydrocolloidcomposedof-o-1,3and-o-1,4galactoseresiduesthataresulfatedatupto40of
thetotalweight,strongnegativechargeovernormalpHrange,associatedwithammonium,calcium,
magnesium,potassium,orsodiumsalts
Solubility isreadilysolubleincoldorhotaqueoussolution,issolubleinhotsolution,treatmentofaqueous
solutionwithpotassiumionprecipitates-carrageenan
Gelformation doesnotformgels,andformright-handedhelices,potassiumchloridepromotesgelformation
of,calciumionpromotesgelformationof
Metabolism hydrolysisofglycosidiclinkagesatlowerpH,especiallypH3.0,alsodesulfationbysulfatases
Viscosity nearlogarithmicincreaseinviscositywithincreasingconcentration,viscosityoffood-gradecarra-
geenandenedasnotlessthan5cpsat75Cfora1.5solution,viscosityrangesfrom5to800cps
for1.5solutionat75C
Source redalgae,predominantlyaqueousextractionfromChondrus,Gigartina,andvariousEucheumaspecies
Molecular
weight
discrepanciesindenitions,nativecarrageenanreportedtohaveaveragemolecularweightof
1.5x10
6
to2x10
7
,food-gradecarrageenanreportedas100000800000or200000400000,
degradedcarrageenan(poligeenan)hasaveragemolecularweightof2000030000,furcellaranhas
averagemolecularweight2000080000
Properties andcombineeasilywithmilkproteinstoimprovesolubilityandtexture,serveasthickening
agent,emulsifier,stabilizer
Synergistic
eects
withlocustbeangum,increaseingelstrength,otherhydrocolloidsmayalsoaectgelstrengthand
cohesiveness
Concentration
infoodproducts
0.0052.0byweight
Majoruses milkproducts,processedmeats,dieteticformulations,infantformula,toothpaste,cosmetics,skin
preparations,pesticides,laxatives
Veterinarni Medicina, 58, 2013 (4): 187205 Review Article
189
2.2. Properties of carrageenan
Thechemicalreactivityofcarrageenansispri-
marilyduetotheirhalf-estersulfategroupswhich
arestronglyanionic,beingcomparabletoinorganic
sulfateinthisrespect.Thefreeacidisunstable,
andcommercialcarrageenansareavailableassta-
blesodiumpotassiumandcalciumsaltsor,most
Figure1.Chemicalstruc-
tureofcarrageenans
itytemperatureandlowergelstrength.Kappatype
carrageenanhasanestersulfatecontentofabout
25to30anda3,6-AGcontentofabout28to35.
Iotatypecarrageenanhasanestersulfatecontent
ofabout28to30anda3,6-AGcontentofabout
25to30.Lambdatypecarrageenanhasanester
sulfatecontentofabout32to39andnocontent
of3,6-AG(Barbeyronetal.2000).
Review Article Veterinarni Medicina, 58, 2013 (4): 187205
190
commonly,asamixtureofthese.Theassociated
cationstogetherwiththeconformationofthesugar
unitsinthepolymerchaindeterminethephysical
propertiesofthecarrageenans.Forexample,kap-
pa-andiota-carrageenansformgelsinthepresence
ofpotassiumorcalciumionswhereaslambda-car-
rageenandoesnot(Micheletal.1997).Thefunc-
tionalityofcarrageenansinvariousapplications
depends largely on their rheological properties.
Carrageenans,aslinear,water-soluble,polymers,
typicallyformhighlyviscousaqueoussolutions.
Viscosity depends on concentration, tempera-
ture,thepresenceofothersolutes,andthetype
of carrageenan and its molecular weight (Lai et
al.2000).Viscosityincreasesnearlyexponentially
withconcentrationanddecreaseswithtempera-
ture.Carrageenansaresusceptibletodepolymeri-
sationthroughacid-catalysedhydrolysis.Athigh
temperaturesandlowpHthismayrapidlyleadto
completelossoffunctionality(Stanley2011).
3. Pharmacokinetics of carrageenan
3.1. Absorption, distribution, metabolism
and excretion (ADME)
Manypharmacokineticsstudiesconcerningthe
oraladministrationofcarrageenanshavebeencon-
ducted in rats (Carey 1958, Hawkins and Yaphe
1965,DewarandMaddy1970,Grassoetal.1973,
Tomarellietal.1974,Coulstonetal.1975,Pittman
et al. 1976, Chen et al. 1981, Nicklin and Miller
1984,Arakawaetal.1988,Nicklinetal.1988),guin-
ea-pigs(Grassoetal.1973,EngsterandAbraham
1976),rabbits(Udalletal.1981)andRhesusmon-
keys(Abrahametal.1972,MankesandAbraham
1975,Pittmanetal.1976,Abrahametal.1983).In
groupsoffiveratsthatreceived0.5nativecarra-
geenan(iota-carrageenanfromE. spinosum) or5
degradedcarrageenanfor10days,faecalexcretion
andweightgainweresimilarbetweenthetwopoly-
mers(DewarandMaddy1970),andnativecarra-
geenan(kappa/lambda from C. crispus), untreated
orheat-sterilisedinmilk,wasquantitativelyexcret-
edinthefaecesofrats(Tomarellietal.1974).No
carrageenanwasfoundintheliversofratsfed25
nativecarrageenan(kappa/lambda fromC. crispus
orIridaea crispata)inthedietforonemonth(Chen
etal.1981),ofratsfeddietscontaining1or5
carrageenan(kappa fromGigartina spp.,iota from
E. spinosum) (Coulstonetal.1975),orofratsfed
dietscontaining5Chondrus crispus carrageenan
(kappailambda) for13weeks(Pittmanetal.1976).
Nocarrageenanwasdetectedinthesmallorlarge
intestineofratsfed5nativecarrageenan(iota
fromE. spinosum) (Grassoetal.1973).Nicklinand
Miller (1984) reported that orally administered
carrageenan (type unidentified) of high relative
molecularmasscouldpenetratethemucosalbar-
rierofadultanimalsviatransportbymacrophages
inPeyerspatches.Carrageenandidnotaffectthe
number or distribution of these cells, however,
whenantigenwasadministeredsystematicallyto
carrageenan-fedrats,theantigen-specificantibody
responsewassuppressed.Thisresultsuggestedthat
carrageenaninterfereswithantigenprocessingby
macrophagesandthusmollifiesnormalimmune
function.Analysisofliversamplesfromratsfed
25 native carrageenan ( kappa/lambda from
C.crispus orC. iridaea) inthedietforonemonth
showedthatonlythesecondwasstoredintheliver
intwoanimals,asdeterminedbythepresenceof
gamma metachromaticreactionsitesintheKupffer
cells(Chenetal.1981).Theresultsofanadditional
earlystudysuggestedthatthekappailambda form
ofcarrageenan,preparedbyanon-standardproce-
durefromeitherC. crispus orGigartina stellata,
isnotsignificantlyabsorbedfromtheintestineof
Wistarrats(Carey1958).Twoadditionalstudies
concerning the absorption of carrageenan have
beenreported(Arakawaetal.1988,Nicklinetal.
1988), however, in neither report is the identity
givenofthespeciesfromwhichthecarrageenan
originated.Moreover,inthelatterstudytheformof
carrageenanthatwasusedisunclear(International
FoodAdditivesCouncil1997).Inthefirststudy,
ratsquantitativelyexcretedthecarrageenan (kappa
form)inthefaeces,andithadthesamegelfiltra-
tion distribution pattern as that of the adminis-
tered material. In the latter study, in male PVG
strainratsgivenradiolabelledcarrageenan(iota
form),thereappearedtohavebeensomeuptake
intotheintestinalwall,Peyerspatches,mesenter-
icandcaecallymphnodes,andserum,however,
the method used to radiolabel the carrageenan
withtritiumisquestionable(InternationalFood
AdditivesCouncil1997).Feedingofguinea-pigs
withnativecarrageenan(iota fromE. spinosum)
at 5 in the diet for 2145 days resulted in the
accumulationof36400 pgigofcaecalorcolonic
tissue. The carrageenan was contained in mac-
rophages (Grasso et al. 1973). Food-grade car-
rageenan (kappa from C. crispus, lambda from
Veterinarni Medicina, 58, 2013 (4): 187205 Review Article
191
C. rispus, iota fromE.spinosum) administered
toguinea-pigsasa1solutionindrinking-water
for two weeks was not retained in the caecum
(EngsterandAbraham1976).Itwasreportedin
anabstractthatcarrageenan(typeandspeciesof
originunidentified)waspresentintheliver,stom-
ach,andsmallintestineofnew-bornrabbitsgiven
40 mg native carrageenan orally. Carrageenan
wasnotdetectedinthecardiacorportalblood
4 h after treatment (Udall et al. 1981). Rhesus
monkeys given 1 native carrageenan (kappai
lambda from C. crispus) in drinking-water for
711weeks,withasubsequent11-weekrecov-
eryperiod,showednoevidenceofcarrageenan
storage(Abrahametal.1972).Inanotherstudyon
rhesusmonkeys,notissuestorageofcarrageenan
(kappailambda fromC. crispus) wasfoundwhen
the monkeys were given 1 native carrageenan
inthedrinking-waterfor10weeks(Mankesand
Abraham1975).Monkeysreceivingdailydosesof
500mgikgbwnativecarrageenan(kappailambda
fromC.crispus) for15monthsexcreted12giml
urine.Theconcentrationwasreportedtobeatthe
limitofdetectionofthemethod(Pittmanetal.
1976).Monkeysreceiving50,200,or500mgikg
bwperdaynativecarrageenan(kappailambda
fromC. crispus) orallyfor7.5yearsshowedno
evidenceofstorageintheliverorotherorgans
(Abrahametal.1983).
3.2. Degradation of carrageenans
in the gastrointestinal tract
Althoughnativecarrageenanmaybedegraded
inthegut,thisisprobablyoflimitedtoxicologi-
calsignificance,since,ifnativecarrageenanwere
sufficientlydegradedtocauseulcerationortu-
mourgrowth,thiswouldhavebeendetectedin
feeding studies. Since food-grade carrageenan
doesnothavethesameeffectsasdegradedcar-
rageenan,itiseithernotdegraded,notdegraded
tothesamemolecularmass,ornotdegradedin
thesameway.Itwouldappearthatcarrageenan
isonlypartiallydegraded,thatmostofthedeg-
radationtakesplaceinthestomach,andthatthis
limiteddegradationhasnoeffectonthewallof
thestomach,wherethepHisverylowandacid
hydrolysisundoubtedlyoccurs.Whenakappai
lambda mixture(fromanunidentifiedspecies)
wasincubatedinsimulatedgastricjuiceatpH1.2
and37C,breakdownofglycosidiclinkageswas
lessthan0.1after3h(StancioffandRenn1975).
Breakdownofkappa-carrageenan(unidentifiedspe-
cies)wasabout15 timesgreaterthanthatoftheiota
form,however,theconditionsofhydrolysis(6hat
pH1.0)weremoredrasticthanthosethatoccurnor-
mallyinthestomach,andthepHwouldbeexpected
tobeconsiderablyhigherinafullstomach(Ekstrom
andKuivinen1983).Thereisnoevidencethatcar-
rageenanisdegradedinthelowergut.Incubationof
acarrageenansolutionwiththecaecalcontentsof
ratsforseveralhoursat37Cdidnotalteritsviscos-
ity,suggestingthatthemicrobialfloraoftheratgut
cannotbreakdowncarrageenan(Grassoetal.1973).
Degradationofcarrageenanbyalargenumberof
intestinalbacteriain vitro hasbeenreported,butthe
carrageenanused(ofanunidentifiedformfroman
unidentifiedspecies)contained20reducingsugar,
whichwouldgiveapositiveresultinthetestmethod.
Amongthebacteriaclaimedtobreakdowncarra-
geenanwereKlebsiella pneumonia andEscherichia
coli,however,boththesespeciescanbegrownon
carrageenangel(Epifanioetal.1981).Ifthesebacte-
riahadbeenabletodegradecarrageenan,theywould
haveliquefiedthegelmediumonwhichtheywere
grown(OchubaandvonRiesen1980).Breakdown
offood-gradecarrageenan(kappa-,lambda-,and
kappailambda-carrageenanfromC. crispus andof
iota-carrageenanfromE. spinosum) isolatedfrom
the faeces of guinea-pigs, rats, and monkeys has
beenreported,butthesiteofbreakdownwasnot
determined.Nointestinallesionswereassociated
withthebreakdown.Themolecularmassobserved
(4050 kDa) was not as low as that of degraded
carrageenan(1020kDa)(Pittmanetal.1976).In
anotherstudythedegradationoffood-gradekappa-
andiota-carrageenanwasstudiedunderphysiolog-
icallyrealisticconditionsinanartificialstomach.
Kappa-carrageenanwasnothydrolysedatpH8or
underthesevereconditionsofpH1.2for6h,and
therelativemolecularmassremainedat>200kDa,
withnomorethan20havingamolecularmassof
<100kDa.Itwasconfirmedthatiota-carrageenanis
moreresistanttodegradationthanthekappa form
(Capronetal.1996).Theoriginatingspecieswere
E. Cottonii forkappa-carrageenanandE. spinosum
foriota-carrageenan,however,thisisnotstatedin
thepaper.Thegreaterstabilityof iota-carrageenan
todegradationmayreflecttheconformationofthe
macromoleculeinthemediumused(Ekstromand
Kuivinen1983,Ekstrom1985,InternationalFood
AdditivesCouncil1997).Noevidenceoffermenta-
tionwasseenafterincubationofratcaecalcontents
Review Article Veterinarni Medicina, 58, 2013 (4): 187205
192
Table2.Acutetoxicity
Carrageenan Species Sex Route LD
50
(mgikgbw) Reference
kappailambdafromC. crispus mouse MiF oral 9150440 FoodandDrugResearch
Labs.1971
NR rat NR intravenous >10 Morardetal.1964
kappailambdafromC. crispus rat MiF oral 5400260 FoodandDrugResearch
Labs.1971
kappailambdafromC. crispus hamster MiF oral 6750570 FoodandDrugResearch
Labs.1971
kappailambdafromC. crispus Guinea-pig NR intravenous >10 Morardetal.1964
lambda fromC. crispus
orG. pistallata
Guinea-pig NR intravenous <1 AndersonandSoman
1966
kappailambdafromC. crispus rabbit MiF oral 2640360 FoodandDrugResearch
Labs.1971
kappailambdafromC. crispus rabbit NR intravenous 120(LD
100
) Duncan1965
NR rabbit NR intravenous <50 Morardetal.1964
Iota rat NR oral >5000 Weiner1991
Iota rat NR inhalation >93074(mgim
3
) Weiner1991
Iota rabbit NR dermal >2000 Weiner1991
NR=notreported,MiF=maleandfemale
statedtobekappa/lambda-carrageenanfromGigartina radula (InternationalFoodAdditivesCouncil1997)
withiota-carrageenanfromE. spinosum (Grassoet
al.1973).AstudyinfemaleWistarratsfedcarra-
geenan(typeandoriginunidentied)asaninertpoly-
saccharidedoesnotprovidequantitativemeasures
ofitsdegradability(Elsenhansetal.1981).Feeding
ofthree-week-oldmaleSprague-Dawleyratsforfour
weekswithadietcontaining5iota-carrageenan
originatingfromE. spinosum (InternationalFood
Additives Council 1997) resulted in a signicant
reduction in the bacterial population of the cae-
cum,asassessedbybacterialcountsandtheactiv-
ityofvariouscaecalmicrobialenzymes,however,
theweightsofthecaecalcontentsandthecaecal
wall were increased (Mallett et al. 1984). Similar
eectswereseeninmiceandhamstersfeddietary
iota-carrageenan(originunidentied)(Mallettetal.
1985),however,inneitherstudywasthedegrada-
tionofcarrageenanmeasured.Onthebasisofthe
ratesofevolutionofmethane,hydrogensulphide,
andcarbondioxidefromaslurryofmixedhuman
faecalbacteria,carrageenan(originandtypeuni-
dentied)wasrankedsecondtofourthineaseof
degradationamong15laxativebres(Gibsonetal.
1990).Inastudyof154bacterialspeciescommonly
foundinthehumancolon,carrageenan(originand
typeunidentied)wasoneofthepolysaccharides
mostresistanttofermentation(Salyersetal.1977).
4. Toxicity of carrageenan
4.1. Acute toxicity
AcutetoxicityissummarisedinTable2.
4.2. Short-term studies of toxicity
Groupsoftwomalealbinoratsfed0,5,10,or
20kappa/lambda-carrageenanfromC. crispus
for 10 weeks grew well, with the exception that
50ofthoseatthehighestdosedied(Nilsonand
Schaller1941).Ingroupsofmaleandfemalerats
fed2,5,10,15,or20kappa/lambda-carrageenan
from C. crispus for periods of 23143 days, the
only adverse effect was reduced growth rates at
dietaryconcentrationsof1020(Hawkinsand
Yaphe1965).Noeffectsonappearanceorbehav-
iourwereobservedinmaleandfemaleOsborne-
Mendel or Sprague-Dawley rats fed 5 kappai
Veterinarni Medicina, 58, 2013 (4): 187205 Review Article
193
lambda-carrageenan from C. crispus for nine
months. Bile-duct proliferation was seen in one
maleOsborne-Mendelrat,andreductionoftheliv-
erlobesandcrenationofthemarginswasobserved
inthreefemales(Coulstonetal.1976).Groupsof
12maleand25femaleSprague-Dawleyratswere
fedadietcontaining4processed,heat-sterilised
kappa/lambda-carrageenanforsixmonths.There
wasnoeffectongrowthrate,andthecaecumand
colonwerenormalongrossandmicroscopicex-
amination(Tomarellietal.1974).Additionof5
iota-carrageenanfromE. spinosum tothedietof
10maleWistarratsfor56daysresultedinslight
diarrhoea(Grassoetal.1973).Groupsof10adult
malealbinoguinea-pigsweregiveneitherwater
ora1solutionofundegradediota-carrageenan
fromE. spinosum. After20days,twooffourtreat-
edanimalshadulcerativelesionsinthecaecum,
and the remaining six animals had lesions at 30
days.Thecontrolgroupremainedhealthy(Watt
andMarcus1969).Itwasreportedinabrieflet-
ter that 5 iota-carrageenan in the diet had the
sameeffect(Sharrattetal.1970).Administration
of5iota-carrageenanfromE. spinosum toseven
femaleguinea-pigsinthedietfor56daysresulted
intheformationofmultiplepin-pointcaecaland
colonic ulcerations (Grasso et al. 1973). Groups
ofthreemaleandthreefemaleDanishLandrace
pigswerefed0,50,200,or500mgikgbwperday
ofkappa-carrageenanfromC. crispus for83days.
Nocompound-relateddeathswereseen,andthe
behaviour,appearance,andfeedintakeoftheani-
malsremainednormal.Therewerenosignificant
changesinhaematological,clinical,chemical,or
urinaryparameters.Areasofinfoldingoftheintact
epitheliumwithinfiltrationofthelaminapropria
ofthecolonicmucosabymacrophagesandlym-
phocytes were seen in one pig at 200 mgikg bw
perdayandtwoat500mgikgbwperday,butthese
effectswereconsideredtobereversible(Poulsen
1973).Maleandfemalerhesusmonkeysweregiven
drinking-watercontaining1kappa-carrageenan
fromC. crispus for711weeks.Theanimalsre-
mainedingoodhealth,andtherewasnoevidence
ofanyadverseeffects.Onefemalekilledatseven
weekshadagrosslynormalgastrointestinaltract,
butsomecapillaryhyperaemiaandmucosaloede-
mawereobservedmicroscopically.Amalekilledat
11weekshadnomicroscopicabnormalities.Two
malesandtwofemaleswereallowedan11-week
recoveryperiodandwerethengivencarrageenanat
escalatingoraldosesof501250mgikgbwperday
forupto12weeks.Nogrossadverseeffectswere
observed,andthemicroscopicchangeswerenot
attributedtocarrageenan(Benitzetal.1973).Male
andfemaleinfantbaboonswererearedfrombirth
to112daysofageoninfantformulacontaining0,
1,or5kappailambda-carrageenanderivedfrom
C. crispus. No effect was seen on organ or body
weights, characteristics of the urine and faeces,
grossfindings,haematologicalorclinicalchemical
variables,orthegrossormicroscopicappearance
ofthegastrointestinaltract(McGilletal.1977).
Groupsof10maleand10femaleSprague-Dawley
rats were fed 0 or 5 conventionally processed
iota-carrageenan from E. spinosum and kappa-
carrageenanfromE. cottonii inthedietforperiods
ofover90days.Anadditional10ratsofeachsex
wereassignedtoa28-dayreversibilityphase.The
changesobservedduringthecourseofthestudy
wereattributedbytheauthorstointakeofadiet
withalowernutritionalvaluethanthebasaldiet.
Thepartialreversalofthecaecalweightchanges
duringthe28-dayreversibilityphaseandtheab-
senceofhistopathologicalchangeswouldsupport
thisconclusion(Robbins1997).
4.3. Long-term studies of toxicity
and carcinogenicity
Lifelongadministrationofkappa/lambda-carra-
geenanfromC. crispus orG. mamillosa atconcen-
trationsof0,0.1,5,15,or25inthediettogroups
offivemaleandfivefemalemiceoftwouniden-
tifiedstrainshadnoadverseeffects(Nilsonand
Wagner1959).Lifetimeadministrationofkappai
lambda-carrageenanfromC. crispus orG. mamil-
losa at concentrations of 0, 0.1, 5, 15, or 25 in
thediettogroupsoffivemaleandfivefemalerats
oftwounidentifiedstrainsresultedinevidenceof
hepaticcirrhosis,onlyatthe25concentration,
with no effect on mortality (Nilson and Wagner
1959).Groupsof30maleand30femaleMRCrats
werefed0.5,2.5,or5kappa-carrageenanfrom
C. crispus inthedietforlife,100malesand100
femalesconstitutedthecontrolgroup.Animalsoc-
casionallydevelopedsoftstoolconsistency,par-
ticularlynearthestartoftheexperiment.There
wasastatisticallynon-significanttrendtowards
an increased incidence of benign mammary tu-
moursandtesticularneoplasmsinthegroupfed
2.5(Rustiaetal.1980).Groupsof15maleand
female Sprague-Dawley rats were given extracts
Review Article Veterinarni Medicina, 58, 2013 (4): 187205
194
ofkappa-carrageenanfromHypnea musciformis
orIrideae crispata ataconcentrationof1or5
inthedietforoneyear.Weightloss(P =0.05)was
observedinalltreatedratsascomparedwiththe
controlgroup,whichreceivedalphacel.Thelivers
ofratsat1werenormalongrossandmicroscopic
examination.Grossandmicroscopicexaminations
oftheliversofratsgiven5kappa-carrageenan
fromH. musciformis werenormal,exceptfornod-
ulesintwoof12livers.Grossobservationofthe
liversofratsreceiving5kappa-carrageenanfrom
I. crispata showeddecreasedsize,roughsurface,
andvascularisationin10i13rats,whichwasprob-
ablyrelatedtotreatment.Microscopically,these
livers were normal, except for focal necrosis in
1 of10livers.Therewasnoevidenceofstorageof
carrageenan-likematerial(metachromatic)inthe
livercellsofanyofthetreatedrats,andnofibrillar
materialwasobservedbyelectronmicroscopy.No
changeswereobservedinthestoolsofratsreceiv-
ing1ofeithercarrageenan,butfemaleratsgiven
5kappa-carrageenanfromI. crispata andmales
giveneithercarrageenanatthe5concentration
hadloosestools.Bloodwasfoundsporadicallyin
thestools,butthefrequencywasnotsignificant
(Coulstonetal.1975).Nineteenmaleand21female
rhesusmonkeyswerefed0,50,200,or500 mgikg
bwkappa/lambda-carrageenanbygavagedailyon
sixdaysaweekforfiveyearsandcarrageenanincor-
poratedintothedietforafurther2.5years.Loose
stools,chronicintestinaldisorders,poorappetite,
andemaciationwereseeninanapparentlyrandom
distribution.Stoolconsistencywasdecreasedina
dose-relatedtrendovertheentire7.5yearsofthe
study,andfindingsoffaecaloccultbloodwerein-
creasedinasimilarfashion.Meansurvivaltimewas
similarinallgroups,andnogrossormicroscopic
changes were detected in the tissues examined.
Sporadicdifferencesinbodyweightfromcontrols
wereseenrandomly,femaleshadsignificantbody-
weightdepressioninthelast2.5yearsofthestudy,
whichdidnotappeartobedose-related.Nocon-
sistent,statisticallysignificantchangesoccurredin
haematologicalorclinicalchemicalvalues,absolute
organweights,ororgan-to-bodyweightratiosaf-
ter7.5yearsoffeedingcarrageenan.Cytochemical
andultrastructuralobservationsrevealednostor-
ageofcarrageenan-likematerialinliversobtained
atbiopsyorinotherorgansobtainedatnecropsy
frommonkeysgivencarrageenan,andnodose-re-
latedgrossormicroscopicchangesinothertissues
(Abrahametal.1983).
5. Biological activity of carrageenan
Sulphatedpolysaccharidesfrommarinealgaecan
havediversebiologicalandactivitiesincludingim-
munomodulatory,anticoagulant,antithrombotic,
antiviralandantitumoreects.Ithasbeensuggested
thatthesenegativelychargedmolecules,including
thesulphatedpolysaccharides,exerttheirinhibitory
eectbyinteractingwiththepositivechargeson
thevirusoronthecellsurfaceandtherebyprevent
thepenetrationofthevirusintothehostcells.It
hasbeenreportedthatcarrageenanhasnoeecton
virusattachmentorpenetrationintohostcells,but
thatthesynthesisofviralproteinsinsidethecells
wasinhibited.Carrageenanhasbeenreportedto
haveanti-HIVactivity,butitsstronganticoagulant
activityisconsideredtobeanadversereactionwhen
usedasatherapeuticdrugforAIDS.
5.1. Carrageenan-induced paw oedema
Carrageenan-inducedratpawoedemaisawidely
usedtesttodetermineanti-inammatoryactivity
andconstitutesasimpleandroutineanimalmodel
forevaluationofpainatthesiteofinammation
withoutanyinjuryordamagetotheinamedpaw
(Sugishitaetal.1981,Henriquesetal.1987,Jainetal.
2001,Paschapuretal.2009,Peterssonetal.2001,Sini
etal.2010).Mousepawoedemahasbeenincreas-
inglyusedtotestnewanti-inammatorydrugsas
wellastostudythemechanismsinvolvedininam-
mation.Intheliterature,thereareabout400papers
reportingtheuseofmousepawoedema(Posadas
etal.2004).Afreshlypreparedsolutionof13
carrageenaninsalineasanintraplantarinjectionin
dosesof50150liscommonlyused(Salveminiet
al.1996,HandyandMoore1998,Nanteletal.1999,
Botting2000,Rosenetal.2000,Jainetal.2001,Guay
etal.2004,Posadasetal.2004,Naudeetal.2010,
Estakhretal.2011),higherconcentrationshavebeen
usedforthemodellingofspecicpathophysiological
conditions(Radhakrishnanetal.2004,Portoetal.
2010,Silvaetal.2010).
Tedevelopmentofoedemaintherathindpaw
followingtheinjectionofcarrageenanhasbeende-
scribedasabiphasic,age-weightdependentevent
inwhichvariousmediatorsoperateinsequenceto
producetheinammatoryresponse.Terearesev-
eralmediatorsinvolvedininammation.Histamine,
serotonineandbradykininaretherstdetectable
mediatorsintheearlyphaseofcarrageenan-induced
Veterinarni Medicina, 58, 2013 (4): 187205 Review Article
195
inammation,prostaglandins(PGs)areinvolvedin
theincreasedvascularpermeabilityandaredetect-
ableinthelatephaseofinammation.Localandior
systemicinammationisassociatedwithenhanced
levelsofthepro-inammatorycytokinesTNF-,
IL-1,andIL-6(Cuzzocreaetal.1999).Teinitial
phase of oedema, which is not inhibited by non-
steroidalanti-inammatorydrugs(NSAIDs)suchas
indomethacinoraspirin,hasbeenattributedtothe
releaseofhistamine,5-hydroxytryptamine(5-HT)
andbradykinin.Tesecondacceleratingphaseof
swellinghasnotonlybeencorrelatedwiththeelevat-
edproductionofprostaglandins,butmorerecently
hasbeenattributedtotheinductionofinducible
cyclooxygenase(COX-2)inthehindpaw(Nantelet
al.1999).ItcanbeblockedbytheNSAIDs(Handy
andMoore1998).Localneutrophilinltrationand
activationalsocontributetothisinammatoryre-
sponsebyproducing,amongothermediators,oxy-
gen-derivedfreeradicalssuchassuperoxideanion
(O
2

)andhydroxylradicals(Salveminietal.1996,
Posadasetal.2004).
Another important mediator in acute inflam-
mationisnitricoxide(NO)whichisproducedin
pathologicalconditionsbythreedistinctisoforms
of nitric oxide synthase (NOS): endothelial NOS
(eNOS), neuronal NOS (nNOS) and inducible
NOS(iNOS).Carrageenancausestheproduction
andreleaseofNOattheinjuredsite.Perfusionof
anon-selectiveNOSinhibitor,NGmonomethyl-i-
arginineacetate(L-NMMA),whichexhibitssome
selectivityforinhibitionofneuronalandendothelial
isoforms,suppressedthereleaseofNOfollowing
carrageenaninjectioninthisstudy.Perfusionofan
inducibleNOSinhibitor,aminoguanidinehemisul-
fate(AG),suppressedthereleaseofNO2.58haf-
tercarrageenaninjection.Neurectomycompletely
suppressedNOreleaseforupto3handpartially
suppressedNOrelease4.58haftercarrageenan
injection.TesendingsindicatethatnNOScon-
tributestotheNOproductioninboththeearlyand
latephase,andthatiNOSonlycontributestothelate
phase.TeproductionandreleaseofNObythese
NOSs are thought to contribute to tissue injury-
andinammation-inducedoedemaandhyperalgesia
(HandyandMoore1998,Omoteetal.2001).
5.2. Intestinal inflammation
WattandMarcus(1971)describedasimplemeth-
odforinducingtheformationofulcersinthelarge
intestineoftheguineapigrequiringonlythead-
ditionofadegradedcarrageenantothedrinking
water.Themethodmaybeusedasanexperimen-
talmodelforthestudyofvariousaspectsofthe
pathologyofulcerativelesionsinthispartofthe
alimentarytract.Freshlyprepared,degradedcarra-
geenanwasaddedtothedrinkingwaterforguinea
pigs to a concentration of 5 . No greater than
2 gikgbodyweightofdegradedcarrageenaninthe
drinkingfluidfor20to45daysresultsinulcerative
lesionsassociatedwithclinicalandpathological
changeswhichincertainrespectscloselyresemble
ulcerativecolitisinman.Clinicallytherewaslossof
weight,loosestools,andoccultorvisiblebloodin
thefaeces.Pathologically,ulcerationwasfoundin
allpartsofthelargebowel,extensivelesionsoccur-
ringintherectum.Microscopically,thesimilarities
includefocalmucosalhaemorrhagesandcellular
infiltrates,oedema,cryptabscesses,irregulardila-
tationofcryptswithlossofmucin-secretingcells
anddegenerationoftheliningepithelium,ulcera-
tioninvolvingmainlythemucosa,aswellasulcera-
tionsinvariousstagesofprogressionandhealing.
Theulcerativelesions,however,appeartobeginin
thecaecumandextenddistallytowardtherectum.
5.3. Anticoagulant and antithrombotic
activity
The blood coagulation system consists of in-
trinsic and extrinsic pathways, with a series of
factorsinvolveinthemechanisms.Bloodcoagu-
lationispromotedbycoagulationfactorsinorder
tostoptheflowofbloodthoughtheinjuredves-
selwallincasesofabnormalvascularconditions
andexposuretonon-endothelialsurfacesatsites
of vascular injury. As endogenous or exogenous
anticoagulantsinterferewithcoagulationfactors
byinactivatingthemorrestrictingtheiractivity,
blood coagulation can be prolonged or stopped
(Wijesekaraetal.2011).Carrageenanisclassified
intovarioustypes,allcontaining2235sulphate
groups.Manyreportsexistontheanticoagulant
activityofcarrageenan.Amongthecarrageenan
types,-carrageenanhasapproximatelytwicethe
activity of unfractionated carrageenan and four
timestheactivityof-carrageenan.However,the
mostactivecarrageenanhasapproximatelyone-
fifteenththeactivityofheparin.Theprincipalbasis
oftheanticoagulantactivityofcarrageenanappears
to be an anti-thrombic property. -Carrageenan
Review Article Veterinarni Medicina, 58, 2013 (4): 187205
196
showed greater anti-thrombic activity than
-carrageenanprobablyduetoitshighersulphate
content, whereas the activity of the unfraction-
ated material was somewhere between the two.
-Carrageenan consistently prolonged the clot-
tingtimeandwasmoretoxicthan-carrageenan.
The difference in sulphate content between the
twocarrageenansdidnotcorresponddirectlyto
differencesinanticoagulationactionandtoxicity
(Shanmugam and Mody 2000). As stated above,
themechanismunderlyingtheanticoagulantactiv-
ityofcarrageenaninvlovesthrombininhibition.
Amidolyticstudiesinitiallyindicatedthatthean-
ti-thrombinactivitymightbemediatedviaAT-III
(anti-thrombin-III),themajormechanismbywhich
heparin acts. In these studies, carrageenans ap-
pearedtoinhibitamidolysisofthrombindirectly
andviaAT-III,however,onlyAT-IIIpotentiated
X
a
amidolysiswasobserved(Kindnessetal.1979).
Theseinteractionsmaybeinfluencedbycertain
criticalqualitiesofthepolyanionicpolymers,i.e.,
sulphation,size,patternofionicsubstitutionand
polymerrigidity(Kindnessetal.1979).However
subsequentstudiesusingAT-III-depletedplasma
showedresidualanti-thrombinactivityinthepres-
ence of carrageenans. -Carrageenan has been
showntopotentiatetheinactivationofthrombin
byanti-thrombinBM.Theseobservationswould
thereforeimplythatthereiseitheranti-thrombin
potentiationviaheparinco-factorII(HC-II)andi
or a direct anti-thrombin effect (Kindness et al.
1980a,b,Wunderwaldetal.1979).
5.4. Antiviral activity
Carrageenan is a selective inhibitor of several
envelopedviruses,includingsuchhumanpatho-
gens as human immunodeficiency virus, herpes
simplexvirus(HSV),humancytomegalovirus,hu-
manrhinovirusesandothers(Girondetal.1991,
Marchettietal.1995,Carluccietal.1999,Caceres
etal.2000,ZacharopoulosandPhillips1997,Stiles
et al. 2008). Carrageenan acts primarily by pre-
venting the binding or the entry of virions into
cells(Bucketal.2006,Grassaueretal.2008).This
findingisconsistentwiththefactthatcarrageenan
resemblesheparansulfate,anHPVcell-attachment
factor.Carrageenanextractedfromseaweed,isan
exceptionally potent inhibitor of papillomavirus
infectivityin vitro.Itwasfoundtobeactiveagainst
arangeofcommonsexuallytransmittedHPVtypes
thatcancausecervicalcancerandgenitalwarts.
Carrageenanisalsoactivein vitroandinmurine
model systems against other viruses, including
herpessimplexvirusesandsomestrainsofHIV-1
(Gonzalesetal.1987,Babaetal.1988,Lynchetal.
1994,Zeitlinetal.1997,WitvrouwandDeClercq
1997).However,in vitroIC
50
valuesforcarrageenan
inhibitionofherpessimplexvirusandHIV-1in-
fectivityareaboutathousand-foldhigherthanthe
IC
50s
observedforcarrageenaninhibitionofgenital
HPVsin vitro.
Carlucci et al. (1997, 2004) reported that the
-carrageenan and partially cyclised -i-carra-
geenanfromGigartina skottsbergiishowedpotent
antiviraleffectagainstdifferentstrainsofherpes
simplexvirus(HSV)type1andtype2.Similarre-
sultswerereportedbyZacharopoulosandPhillips
(1997)whodescribedtheabilityofcarrageenanso-
lutions(lambda, kappa,oriota)topreventHSV-2
infectionanddeSF-Tischeretal.(2006).
Leibbrandt et al. (2010) tested a commercially
availablenasalspraycontainingiota-carrageenanin
aninfluenzaAmouseinfectionmodel.Treatment
of mice infected with a lethal dose of influenza
APR8i34H
1
N
1
viruswithiota-carrageenanstart-
ingupto48hpostinfectionresultedinastrong
protection of mice similar to mice treated with
oseltamivir. Since alternative treatment options
forinfluenzaarerare,thenasalspraycontaining
iota-carrageenanisanalternativetoneuraminidase
inhibitorsandshouldbetestedfortheprevention
andtreatmentofinfluenzaAinclinicaltrialsin
humans.Ecclesetal.(2010)investigatedtheeffi-
cacyandsafetyofaniota-carrageenannasalspray
in patients with common cold symptoms. Nasal
spraysappeartobeapromisingtreatmentforsafe
andeffectivetreatmentofearlysymptomsofthe
commoncold.
5.5. Anti-tumour and immunomodulatory
activities
Severalstudieshavereportedthatcarrageenans
haveantiproliferativeactivityincancercelllinesin-
vitro,aswellasinhibitoryactivityoftumorgrowth
inmice(Yuanetal.2004,2006,Zhouetal.2004,
2006,YuanandSong2005,RochadeSouzaetal.
2007).Inaddition,theyhaveantimetastaticactivity
byblockingtheinteractionsbetweencancercells
and the basement membrane, inhibit tumor cell
proliferationandtumorcelladhesiontovarious
Veterinarni Medicina, 58, 2013 (4): 187205 Review Article
197
substrates,buttheirexactmechanismsofaction
arenotyetcompletelyunderstood.Yamamotoet
al.(1986)reportedthattheoraladministrationof
severalseaweedscancauseasignificantdecreasein
theincidenceofcarcinogenesisinvivo.Hagiwara
etal.(2001)investigatedthemodifyingeffectsof
carrageenanoncoloniccarcinogenesisinmalerats.
No treated related changes in clinical signs and
body weights were found. Histopathological ex-
aminationdidnotdemonstrateanyenhancement
bycarrageenancarcinogenesis,carrageenandoes
notpossessanypromotingactivityatthehighest
dietarylevelof5.0forcolorectalcarcinogenesis
underthepresentexperimentalconditions.
5.6. Other biological activities
Theantioxidantactivityofallcarrageenanshas
beenstudied.-Carrageenanexhibitedthehigh-
estantioxidantandfreeradicalscavengingactivity.
RochadeSouzaetal.(2007)demonstratedaposi-
tivecorrelationbetweensulfatecontentandantiox-
idantactivity.Thepresentfindingsprovideabasis
forfurtherexperimentsontheidentificationand
characterisationofspecificcompoundswithrela-
tivelyhighantioxidantactivities.Carrageenanfrom
redmarinealgaeisknowntobeapotentinflamma-
toryagentinrodentsandprimesmiceleucocytes
toproducetumournecrosisfactor-(TNF-)in
responsetobacteriallipopolysaccharide.Moreover,
some types of carrageenans induce potent mac-
rophageactivation,whilesomecarrageenansap-
peartoinhibitmacrophagefunctions(Wijesekara
etal.2011).ThefeedingofFischer344ratsondiets
containing15kappailambda-carrageenanfrom
G. radularesultedinacholesterol-loweringeffect
(Reddyetal.1980).Similareffectsoftheinclusion
ofcarrageenaninthedietmayresultinreduced
bloodcholesterolandlipidlevelsinhumansubjects
(Panlasiguietal.2003).
6. Uses of carrageenan
6.1. Industrial uses of carrageenan
Immobilisationofbothenzymesandwholecell
systems is of major importance in the improve-
ment of the stability, activity and reusability of
thesebiocatalysts.Carrageenanisasuitablesup-
portmaterialfortheimmobilisationofwholecells,
asprovenbyseveralapplicationsindifferentindus-
trialprocesses.Theapprovalofcarrageenanasa
food-gradeadditiveandtheeaseoftheimmobi-
lisationprotocolhavepromoteditsapplicationin
thefoodindustries.Themildimmobilisationand
reactionconditionsappliedforcarrageenanim-
mobilisationofwholecellsallowstheirapplication
inhighly(enantio)selectiveproductionprocesses
forpharmaceuticalcompoundsandfinechemicals
(VandeVeldeetal.2002).
6.2. Industrial food applications
Industrialvinegarproductionisabiochemical
processwhichutilisesbacteria.Osugaetal.(1984)
describedtheuseofabubble-mixedreactorand
-carrageenan gel beads as carriers for the con-
tinuousproductionofaceticacid.Animprovement
wasattemptedthroughtheuseofanair-liftreactor
(Mori1993),usingthecultureofAcetobacter species
K1024isolatedfromacommercialvinegarbroth.
Morerecently,asuccessfulcontinuousproduction
ofvinegarwasreportedusingabubblemixedtab-
letopbioreactorwith-carrageenan-immobilised
Acetobacter suboxydanscells(TosaandShibatani
1995).Fermentedmilkproductscanbeobtained
bysimultaneousacidificationandinoculationof
skimmed milk by immobilised mixed cultures.
ThreedifferentstrainsofLactococcus lactisand
onestrainofLeuconostoc mesenteroidesweresepa-
ratelyimmobilisedin-carrageenanilocustbean
gumgel(2.75and0.25wiw,respectively)and
usedina2-Lstirredreactor(Sodinietal.1997a,b).
Mensourandcolleaguedescribedtheimmobili-
sation system of yeast cells (Saccharomyces sp.)
for beer production using -carrageenan beads
continuouslyproducedbyastaticmixerprocess
(Mensouretal.1996,1997).Ethanolproduction
fromglucoseusingcellsofZymomonas mobilisim-
mobilisedin-carrageenanwasinvestigatedina
fluidisedbedfermenter.Thisresearchwasextend-
edtostudytheproductionofethanolfromstarch
(Krishnan1999)usingthebacteriaco-immobilised
withanindustrialglucoamylaseincarrageenangel
beadsandusedinaglasscolumnfermenter.The
carrageenan gel matrix was reported to provide
protectionofimmobilisedSaccharomyces cerevi-
siaecells(Nortonetal.1995)andcontinuousetha-
nolproductionfrompineapplecannerywastewas
attemptedusingtheseyeastcellsimmobilisedin
-carrageenan(Nigam2000).
Review Article Veterinarni Medicina, 58, 2013 (4): 187205
198
6.3. Pharmaceutical applications
6.3.1. Tetracycline and chlorotetracycline
production
Tetracyclines represent one of the most im-
portant groups of antibiotics and the method
normally used for their industrial production is
conventional fermentation. Asanza-Teruel et al.
(1997) used Streptomyces aureofaciens immobi-
lisedin-carrageenaninanattempttoimprove
the production of tetracycline and chlorotetra-
cycline.
6.3.2. Semi-synthetic antibiotic production
Semi-syntheticantibioticsarepreparedviathe
couplingofa-lactamcorewiththeso-calledside
chain,suchasphenylaceticacid,o-phenylglycine,
oro-p-hydroxyphenylglycine.6-Aminopenicillanic
acid(6-APA)isobtainedbytheenzymatichydroly-
sisofpenicillinGproducedbyfermentation.The
suitabilityof-carrageenanasasupportfor6-APA
productionwastestedwithE. colicellswithpeni-
cillin-amidaseactivity(NagalakshmiandPai1997).
Thecellswereimmobilisedwithanefficiencyof
90andcouldbeusedfor20repeatedcyclesre-
taining60oftheinitialpenicillin-amidaseactiv-
ity.Thecarrageenangelbeadswerehardenedwith
gluteraldehyde. Among the side chains, o-p-hy-
droxyphenylglycine is one of the most impor-
tantprecursors,asitisusedforthesynthesisof
amoxicillin and cefadroxil. Recombinant E. coli
cells expressing both dihydropyrimidinase and
carbamoylasewereimmobilisedin-carrageenan
and were able to convert oii-hydroxyphenylhy-
dantointoo-p-hydroxyphenylglycine.Inasingle-
stepreactionaconversionof93wasobtained,
whilea20valuewasobservedwiththestrainof
Agrobacterium radiobacter,whichcontainedthe
original dihydropyrimidinase gene cloned into
E.coli(Chaoetal.1999).
6.3.3. -aspartic acid production
Anumberofo-aminoacidshavebeenshownto
be important intermediates in drug production.
o-Aspartic acid can be used as a component of
syntheticpenicillins(TakamatsuandTosa1993).
Whenoii-asparticacidisusedasasubstratefor
the i-aspartate -decarboxylase of P. dacunhae
cells,i-asparticacidisconvertedtoi-alaninebut
o-aspartic acid remains unchanged due to the
high stereospecificity of the biocatalyst. In this
way o-aspartic acid and i-alanine can be pro-
ducedsimultaneouslyusingP. dacunhaecellsim-
mobilised in carrageenan (Takamatsu and Tosa
1993).oii-Asparticacidischemicallyproduced
fromfumaricacidandammonia.o-Asparticacid
iscrystallisedbyacidificationofthereactorefflu-
entandrecoveredbycentrifugation.i-Alanineis
alsorecoveredbycentrifugationaftercrystallisa-
tionbytheadditionofammoniatotheresulting
liquorfollowedbyconcentrationandcooling.This
systemforthecontinuousproductionofo-aspartic
acidandi-alanineusingP. dacunhaecellsimmo-
bilisedin-carrageenanhasbeenindustrialised
since1988.Industrialapplicationofimmobilised
wholecellswiththeuseof-carrageenangelsfor
thepreparationofi-asparticacidwasfirstdevel-
opedbyChibata(TanabeSeiyaku,Japan)in1973.
The industrial production of a number of other
compounds(i-malicacid,i-alanine,i-tryptophan,
1,5-dimethyl-2-piperidone) for use in food and
medical applications is based on the same prin-
ciples(foradetaileddiscussionseeVandeVelde
etal.2002).
6.4. Cleaning of industrial effluents
An efficient integrated nitrogen removal sys-
tem was developed by the co-immobilisation of
Nitrosomonas europaeaandPseudomonas sp.in
-carrageenan, taking advantage of the oxygen
gradientinsidetheentrapmentbeads(DosSantos
etal.1996a,b).TheimmobilisationofM. aurum
in-carrageenananditsuseinanair-bubblefer-
menterresultedinanimprovementofitsmorpho-
line-degradingcapacity(Swainetal.1991,Poupin
etal.1996).Thecarrageenangelandimmobilised
microbialcellsmethodwasusedfor4-chlorophe-
noldegradation(Wangetal.1997).Aerobicand
anaerobicmicrobialcommunitieshavealsobeen
co-immobilised into -carrageenanigelatin gel
beads(GardinandPauss2001).Underair-limited
conditionstheseimmobilisatescatalysethedegra-
dationof2,4,6-trichlorophenol.Pentachlorophenol
pesticidedegradationincontaminatedsoilwasat-
tempted with the use of Pseudomonas sp. UG30
cellsimmobilisedin-carrageenan(Cassidyetal.
1997).
Veterinarni Medicina, 58, 2013 (4): 187205 Review Article
199
6.5. Other uses of carrageenan
Inthelanguageoffoodchemists,carrageenanis
variablycalledanemulsifier,stabiliser,colloid,or
gum.Manyproductsthatwenowtakeforgranted
especiallysoymilk,chocolateandotherflavoured
milks,dairyproducts,infantformulas,andnutri-
tionalsupplementbeveragesrelyuponcarrageenan
fortheiruniformconsistencies.Theycouldnotbe
made,packagedandstoredforlongperiodsoftime
withoutthisingredient.
Carrageenansareusedtogel,thicken,orsuspend,
thereforetheyareusedinemulsionstabilisation,
forsyneresiscontrol,andforbodying,bindingand
dispersion.Themajorusesareinfoods,particularly
dairyapplications.Furcellarangenerallyfindsap-
plicationssimilartothoseforkappa-carrageenan.
Historically,furcellaranhasdominatedtwomajor
Europeanapplicationfields:tartorcakeglazepow-
dersandflanpowders.Todaythespecialproperties
ofexcellentgeltextureandflavourreleasemake
furcellaranapreferredproductforuseinmilkpud-
dingpowders.Carrageenanisuniqueinitsabil-
itytosuspendcocoainchocolatemilkatverylow
concentrations(ca.300ppm),noothergumhas
been found to match it. A very delicate milk gel
structure,undetectableonpouringordrinkingthe
milk,isbelievedtoholdthecocoainsuspension.A
substantialdifferentialbetweentheconcentrations
atwhichsettlingofcocoaoccursandthatatwhich
visiblegelationisevidentisrequiredforpractical
stabilisation.Thisisachievedbycarefulselection
ofweedtypeandquality.Iota-carrageenanswhich
havetexturesverysimilartothoseofgelatingelsare
usedindessertgelformulations.Theyhaveanad-
vantageovergelatingelsinthattheirmeltingpoint
ishigher,makingthemmoresuitableintropical
climatesorwhererefrigerationisnotavailable.This
is offset to some extent by the different texture,
sincethesegelsdonotmeltinthemouth,asdoes
gelatin.However,afurtheradvantageisthatiota
gelsretaintheirtenderstructureonaging,where-
asgelatintendstotoughen.Thisisimportantfor
ready-to-eatdesserts,popularinEurope.Kappa-
carrageenanorfurcellaranbyitselfisunsatisfac-
toryfordessertgelapplicationsduetotheshort,
brittlestructureofitsgel.Thiscanbeameliorated
bytheincorporationoflocustbeangalactoman-
nanintotheformulation,andkappa-locustbean
oriota- kappa-locustbeanblendsarealsooffered
forthisapplication.Toachievesparkling-cleargels
itisnecessarytousealocustbeangumwhichhas
been clarified by filtration. The clarified gum is
producedforthispurposebyseveralofthemajor
carrageenanmanufacturers.
Intoothpastescarrageenansfunctionasabind-
ertoimpartthedesiredrheologicalpropertiesto
the paste and to provide the cosmetic quality of
sheen.Toothpastesconsistofingredientswhich
interact in complex and in poorly understood
waysandthecarrageenanoftenmustbecarefully
tailoredtoachievesatisfactoryperformanceina
particularformulation.Carrageenansufferssevere
competitionintheU.S.domesticmarketfromsodi-
umcarboxymethylcellulose,amuchcheapergum.
Despitethis,businesshasbeenretainedandre-
gainedduetothesuperiorqualityandappearance
carrageenanimpartstotoothpastes.Outsidethe
UnitedStatescarrageenanhasmaintainedastrong
positioninthisapplication,due,amongotherfac-
tors,toitsimmunitytodegradationbyenzymes
whichattackcellulosegums.Gelatinousextracts
oftheChondrus crispus(IrishMoss)seaweedhave
beenusedasfoodadditivesforhundredsofyears.
Carrageenanisavegetarianandveganalternative
togelatin.
Whenusedinfoodproducts,carrageenanhasthe
EUadditiveE-numberE407orE407awhenpresent
asprocessedeucheumaseaweed,andiscommonly
usedasanemulsifier.InpartsofScotland(where
it is known as (An) Cairgean in Scottish Gaelic)
and Ireland (variety used is Chondrus Crispus
knowninIrishGaelicvariouslyascarraign[lit-
tlerock],fiadhain[wildstuff ],cluimhincait[cats
puff ], mathair an duilisg [mother of seaweeds],
ceanndonn[redhead]),itisknownasCarrageen
Moss.Itisboiledinmilkandstrained,beforesugar
andotherflavouringssuchasvanilla,cinnamon,
brandy,orwhiskyareadded.Theend-productis
a kind of jelly similar to pannacotta, tapioca, or
blancmange.Wheniota-carrageenaniscombined
withsodiumstearoyllactylate(SSL),asynergistic
effectiscreated,allowingforstabilisingiemulsify-
ingnotobtainedwithanyothertypeofcarrageenan
(kappailambda)orwithotheremulsifiers(mono
anddiglycerides,etc.).SSLcombinedwithiotacar-
rageenaniscapableofproducingemulsionsunder
bothhotandcoldconditionsusingeithervegetable
oranimalfat.
Theuseofcarrageenaninthefoodindustryisof-
tendiscussedinthecontextofitssafety.Authorities
worldwidesuchasJECFA,ScientificCommunity
onFood(SCF),andInternationalFoodAdditives
Council (IFAC) have extensively evaluated the
Review Article Veterinarni Medicina, 58, 2013 (4): 187205
200
safetyofcarrageenan.Incontrasttothefindings
presentedbyTobacman(2001)andTobacmanet
al. (2001) all of these authorities agree that car-
rageenan is safe for use in foods. In 1978, the
ScientificCommitteeforFood(SCF)endorsedthe
AcceptableDailyIntake(ADI)of075mgikgbw
establishedforcarrageenanbytheJointFAOiWHO
ExpertCommitteeonFoodAdditives(JECFA1974,
SCF1978).
7. CONCLUSION
Carrageenansarecommerciallyimportanthy-
drophilic colloids (water-soluble gums) which
occurasmatrixmaterialinnumerousspeciesof
redseaweeds(Rhodophyta)whereintheyservea
structuralfunctionanalogoustothatofcellulose
inlandplants.Chemicallytheyarehighlysulfated
galactans.Duetotheirhalf-estersulfatemoieties
theyarestronglyanionicpolymers.Thechemical
reactivityofcarrageenansisprimarilyduetotheir
half-estersulfategroupswhicharestronglyani-
onic,beingcomparabletoinorganicsulfateinthis
respect.Thefreeacidisunstable,andcommercial
carrageenansareavailableasstablesodiumpotas-
sium and calcium salts or, most commonly, as a
mixtureofthese.Theassociatedcationstogether
with the conformation of the sugar units in the
polymerchaindeterminethephysicalproperties
of the carrageenans. The functionality of carra-
geenans in various applications depends largely
ontheirrheologicalproperties.Viscositydepends
on concentration, temperature, the presence of
other solutes, and the type of carrageenan and
its molecular weight. Viscosity increases nearly
exponentially with concentration. Most of the
toxicological studies in which an identifiable
typeofcarrageenanandanidentifiableseaweed
specieswereusedwereundertakenwithkappa-
or kappailambda-carrageenan from C. crispus.
Theresultsofthefewparallelstudiessuggestthat
therearenolargedifferencesintheeffectsofthe
different forms of carrageenan or in the effects
ofcarrageenanspreparedfromdifferentspecies
of seaweed. Carrageenans have very low toxic-
ity, and have been shown not to be teratogenic.
Carrageenanisusedinmanydairyproductssuch
ascreamcheese,cottagecheese,skimmedmilk,
andyogurtaswellasdessertsandsweetssuchas
custards,icecream,milkshakes,piefillingsand
chocolateproducts.Inadditiontouseasafood
additive,carrageenanisalsousedinairfreshen-
ergels,toothpaste,firefightingfoam,shampoo,
cosmeticcreamsandshoepolish.Inbiotechnol-
ogy, carrageenan is used as a gel to immobilise
cellsienzymes. A special use of carrageenan is
inexperimentalmedicineforthetestingofanti-
inflammatorydrugs.
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Received:20121121
Acceptedaftercorrections:20130409
CorrespondingAuthor:
LenkaBartosikova,PalackyUniversity,FacultyofMedicineandDentistry,DepartmentofPhysiology,Hnevotinska3,
77515Olomouc,CzechRepublic
Tel.+420585632361,E-mail:bartosil@tunw.upol.cz