GlobalResearchjournalofNaturalScience&Technology(GRJNST)Volume:04-Issue2(2026),2071ISSNP:2790-7643ISSNE:2790-7651www.grjnst.nethttps://doi.org/10.53762/grjnst.04.02.022BiochemicalcharacterizationandprofilingofmajorseedstorageproteinsinselectedmedicinalplantfoundinSindhPakistanReceived:25March2026Accepted:10March2026.Published:27April2026S.K.KhanzadaInstituteofPlantSciences,UniversityofSindhsamina.khanzada@usindh.edu.pkAKKhanzadaInstituteofPlantSciences,UniversityofSindhamina.kabir@usindh.edu.pkBeenishkhanzadaInstituteofBiochemistryUniversityofSindhbeenish@usindh.edu.pkShaheenMemonInstituteofPlantSciences,UniversityofSindhShaheen.memon1985@gmail.comB.SadiqInstituteofPlantSciences,UniversityofSindhSadique.khiliji@scholars.usindh.edu.pkImranAliBhurtInstituteofPlantSciences,UniversityofSindhemranali007007@gmail.comAbstract:Inthisstudyinvestigatedthecompositionandbiochemicalpropertiesofseedstorageproteinsinimportantmedicinalplants,includingtheseedsofSyzygiumcuminiL.,LawsoniainermisL.,Calotrpisprocera.RBro,RicinuscommunisL.,LinumusitatissimumL.,BrassicajunceaLCzern.,WithaniacoagulansDun,andCucumismelovar.agrestisNaudin.Proteinextractionfromseedflourwascarriedoutusingsequentialstepsofextraction:delipidation(removalofoil),water(albumin),5.0MNaCl(globulin),70%ethanol(prolamin),and150.2Msodiumphosphatebuffer,pH8.0(glutelin).QuantificationwasperformedusingthedyebindingmethodofBradfordwhichshowedsubstantialdifferencesintheirconcentrationsandtotalyields.Comparativeanalysisofseedstorageproteinsshoweddifferentdominancepatternsamongtheplantsstudied.ThehighestlevelsofalbuminwerefoundinBrassicajunceaLCzern.(50.40%),whichsuggestsastrongwater-solubleproteinfraction,thehighestglobulincontentwasfoundinRicinuscommunis(67.11%),suggestingahighsalt-solubleproteinfraction,thehighestprolamincontentwasfoundinWithaniacoagulans(55.93%),suggestingastrongalcohol-solubleproteinfraction,andthehighestglutelinwasfoundinSyzygiumcumini(15.12%).Keywords:Seedstorageproteins(SSPs)Extraction,Proteinestimation,MedicinalplantsINTRODUCTIONDuringseedgerminationandtheearlystagesofseedlinggrowth,seedstorageproteinsserveasimportantstoresofnitrogenandaminoacids.Theseproteinsareclassifiedbasedontheirsolubilityandstructuralpropertiesintoalbumins,globulins,prolamins,andglutelins.Inadditiontotheirphysiologicalroleinplants,seedstorageproteinshavebecomemoreinterestingtostudyduetotheirnutritionalvalueandbioactiveactivities,suchasantibacterial,antioxidant,andmedicinalproperties(Yangetal.,2023;Zengetal.,2024),Recentstudieshavealsoemphasizedtheimportanceofmedicinalplantseedsasasourceofbioactivepeptidesandfunctionalproteinswithpharmacologicalapplications(Latif&Nawaz,2025).TheseedsofSyzygiumcuminiL.richinalbuminsandglobulinsandarewellknownfortheirantioxidantandantidiabeticactivities,whichareattributedtoavarietyofbioactivechemicalsandproteinfractions(Latif&Nawaz,2025).Thepresenceofhighlevelsofstorageproteins,especiallyalbumins,anditshepatoprotectiveandantibacterialpropertiesmakeLawsoniainermisL.animportantplantintraditionalmedicine(Kumaretal.,2024).Anumberofproteincomponents,includingdefense-relatedandenzymaticproteins,haveanti-inflammatoryandantibacterialpropertiesintheseedsofCalotropisproceraR.Br.(Tiwarietal.,2022).RicinuscommunisL.seedscontainbothtoxicproteinssuchasricinandstorageproteinssuchasglobulins,buttheseedscanbeusedmedicinallyifproperlyprepared(Yangetal.,2023).TheseedsofLinumusitatissimumL.containglobulinsandalbuminsandareusedmedicinallyfortheircardioprotective,antioxidant,andanticanceractivities,inpartduetothecompositionoftheirproteinsandrelatedmetabolites(Zengetal.,2024).Duetotheirintricatebiochemicalcomposition,BrassicajunceaLCzern.,seedscontainantibacterialandchemoprotectivecompoundsandhighlevelsofglobulins(Yangetal.,2023).Researchonstorageproteinalbumin,globulin,prolamin,andglutelin.ofApiaceaeFamilymedicinalplant,seedsarevaluable,nutrientsourcesofseedproteins,theseseedsareutilizedinhomeremediestomanagegastrointestinalissuessuchasindigestion,flatulence,andcolicpain,whilealsoactingasflavoring,aromatic,andpreservativeagentsinfoodsupplies(S.K.Khanzadaetal.2021),BioactiveproteinsandsecondarymetaboliteslikewithanolidesarereportedtoberesponsiblefortheantidiabeticandimmunomodulatorypropertiesofWithaniacoagulansDun.seeds(Latifetal,2025).Storageproteinsarealsoimportantfromanutritionalstandpoint,andseedsofCucumismelovar.agrestisNaudin.provideantioxidantandanti-inflammatorypropertiesfortraditionaltherapeuticuse(Kumaretal.,2024).Theyhavebeeninvestigatedasasourceoffoodandpharmaceuticalsduetotheirbiochemicalcomposition,nutritionalimportance,andmedicinalproperties,includingmedicinalplantseedstorageproteins(M.Kumaretal.,2022).Inhumanmetabolism,seedstorageproteinsexhibitawiderangeofactivities,andalbuminsandglobulinsareabundantinangiospermicplants,includingmonocotsanddicots,suchascereals,ferns,andpalms(Templemanetal.,1987).Glutelins,ontheotherhand,areresistanttopepsindigestionandthuslessdigestiblebyhumans.Thebiochemicalcharacterizationof(SSPs)inmedicinalplantsisimportantforunderstandingtheirnutritionalvalueandtherapeuticapplicationsinthefoodandpharmaceuticalindustries.SSPsaresimplebiomoleculesthatarenecessaryforthegrowth,development,andreproductionofplants.(López-Guzmánetal.,2025).Inthisstudyinvestigatedthebiochemicalpropertiesandproteinprofilesofseedstorageproteinsineightmedicinallyimportantplantspeciesofvariousfamilies.MATERIALANDMETHODSMaterial:Seedsofeightmedicinallyimportantplantsbelongingtothedifferentplantfamilies:SyzygiumcuminiL.,(Myrtaceae),LawsoniainermisL.,(Lythraceae),Calotrpisprocera.RBro(Asclepiadaceae),RicinuscommunisL.,(Euphorbiaceae),LinumusitatissimumL.,(Linaceae)BrassicajunceaLCzern.,(Brassicaceae),WithaniacoagulansDun(Solanaceae),andCucumismelovar.agrestisNaudin.(Cucurbitaceae).werecollectedfromthreedifferentlocalities,Nawabshah,Hyderabad,JamshoroUniversityofSindhJamshoroforconductedtoinvestigatethecomposition,biochemicalpropertiesofseedstorageproteinsinimportantmedicinalplants.Seedstorageproteinextraction:Theseedweregroundwithmortarpistil,theflourwasdelipidatedbysolventextractionbystirringwithpurehexane(1:10w/v)for30min,andthisprocedurerepeatedthreetimesatroomtemperature.Usingthemicroextractionmethod(Sammouretal.,1999),thesamplewasairdriedandkeptunderhooduptotwenty-fourhoursinalcohol,water,highsalt,andinalkalinebufferconditions.Finally,theseedwasextractedintriplicateinEppendorftubeswith500micrograms(i)deionizedwater(ii)145.0MNaCl(iii)70%ethanoland(iv)0.2M,Na₃PO₄buffer,pH8.0,toextractthefourmajorseedstorageproteins(prolamin,albumin,glutelin,andglobulin),eachat500micrograms,for20minat350rpm/25CusingThermomixercomfort(Eppendorf,Germany).Thisextractionwasperformedat14000rpmfor15minat4ºC(BiofugePrimoRHeraeus,Japan)andrepeatedtwicetoremoveresidualproteinfromeachfraction.Thesupernatantsofeachfractionwerepooledandstoredat-20Cuntilproteinprofiling.Thetotalproteinfromtheentireextractionwasmeasuredusingthemodifiedcolorbindingassay(Bradford,1976)withbovineserumalbuminasastandard.Proteinquantitationwasperformedintriplicateusingamicroplatereader(SunriseTecan,Austria).ResultsandDiscussionTheseedstorageproteincontentofeightmedicinalplantspeciesfromSindh,PakistanispresentedinTable1.Theresultsshownoticeablevariationamongthespeciesinthefourmainproteinfractions,namelyalbumin,globulin,prolamin,andglutelin.Inthisstudyinvestigatedthebiochemicalpropertiesandproteinprofilesofseedstorageproteinsineightmedicinallyimportantplantspeciesofvariousfamilies.S.cuminiL.,L.inermisL.,C.procera.RBro,R.communisL.,L.usitatissimumL.,B.junceaLCzern.,W.coagulansDun,andC.melovar.agrestisNaudin..Thesedifferencesindicatethateachspeciesiscomposedofadifferentproteinset,whichcouldmakeitmoreorlessnutritiousandplaydifferentbiologicalroles.Albuminshowedawiderangeofvalues(22.88to50.40),withB.junceaLCzernhavingthehighestcontent(50.40),followedbyC.procera.RBro,(44.40),L.inermisL.,(41.86),L.usitatissimumL.,(40.14),andW.coagulansDun(31.40),andthelowestinR.communisL(22.88).ThishighalbuminproportioninB.junceaLCzernindicatesitmaybeagoodsourceofwater-solubleproteinswithhighdigestibility.Globulinvaluesvariedconsiderably,from1.70%inL.inermisL.to67.11%inR.communisL.,withC.procera.RBro,(36.02%)andL.usitatissimumL.,(34.82%)alsoexhibitingsignificantglobulinlevels.Inapreviousstudy,43.25%proteinrecoveryand17.2%extractionefficiencywasreportedforL.usitatissimumL.,proteinextractedatpH12(Sharmaetal.,2022).However,theproteinfractionsdifferedandthemajorseedstorageproteinswerealbumin(40.14%),globulin(34.82%),prolamin(22.38%),andglutelin(11.57%).L.inermisL.,(1.70%)andW.coagulansDun(4.29%)containedthelowestamounts.Thesedifferencesrepresentspecies-specificstoragestrategiesandcanimpactseednutritionalqualityandindustrialapplications.Prolaminwasthemajorproteinfractioninmanyspecies,W.coagulansDun(55.00%),L.inermisL.,(53.32%),andS.cuminiL(48.00%)followedbyCucumismelovar.agrestis(39.80%),B.junceaLCzern(26.56%),R.communisL(5.16%),andC.procera.RBro,(7.75%).Thehighprolamincontentincertainspeciessuggestspotentialfunctionalrolesinseedphysiologyandpossibleapplicationsinfoodandpharmaceuticalindustries,althoughprolaminsaregenerallylessnutritionallybalancedduetolowlysinecontent.Glutelincontentrangedfrom3.10%to15.12%,withS.cuminiL(15.12%)andC.melovar.agrestisNaudin(14.42%)showingthehighestlevels.ModerateamountswereobservedinC.procera.RBro,(11.81%)andL.usitatissimumL.,(11.57%),whereasL.inermisL.,(3.10%)andB.junceaLCzern(3.43%).Thedistributionoftheseproteinsindicatesthatalbuminandprolaminarethemajorproteinsinmostspecies,andthatglobulinisthemajorproteininR.communisL.,(SSPs)storageproteincompositionishighlightedbytheobservedvariationinproteinfractions,whichmaybeimpactedbygeneticdeterminants,ecologicaladaptability,andseedfunctionalneeds.Theextenttowhichtheproportionsoftheseproteinsdifferbetweenspeciesmayhavesignificantnutritionalqualityandindustrialapplications.Table1.ComparisonoftheconcentrationsandtotalpercentyieldofthemajorseedstorageproteinsfromSyzygiumcuminiL.,LawsoniainermisL.Calotrpisprocera.RBroRicinuscommunisL.,LinumusitatissimumL.,BrassicajunceaLCzern.,WithaniacoagulansDun,andCucumismelovar.agrestisNaudin.seedsfoundinSindh,Pakistan.S.NO.PlantNameFamilyNameAlbuminmg/gGlobulin*#Prolamin*%yieldmg/g%yield#mg/g%yieldGlutelin*#mg/g%yield1.SyzygiumcuminiL.Myrtaceae0.8330.530.197.001.3048.000.4015.122.LawsoniainermisL.Lythraceaea0.6141.860.021.700.7853.320.043.103.Calotrpisprocera.RAsclepiadaceace0.1344.400.1036.020.027.750.0311.81Bro4.RicinuscommunisL.Euphorbiaceae1.8622.885.4767.110.425.160.394.815.LinumusitatissimumLinaceae1.3440.141.1734.820.7522.380.3811.57L.6.BrassicajunceaLBrassicaceae0.7650.400.2919.600.4026.560.053.43Czern.7.WithaniaSolanaceae0.0831.440.014.290.1555.000.029.32coagulanceDun.8.CucumismeloL.var.CucurbitaceaeagrestisNaudin.0.0330.770.0115.000.0539.800.0114.43*Concentrationsinmg/gofseedflour.Valuesaremeanofthreeindependentextractions.#%yieldofaparticularproteinintotalproteincontentsofseedflour.Table2:Seedstorageprotein(SSPs)composition,functionalimportance,andmedicinalsignificanceofselectedmedicinalplantsfoundinSindhPakistanMedicinalplantsWithImportanceofproteinMedicinalsignificancefamilynameSyzygiumcumini(Myrtaceae)Lawsoniainermis(Lythraceae)Calotropisprocera(Asclepiadaceae)Ricinuscommunis(Euphorbiaceae)Linumusitatissimum(Linaceae)Brassicajuncea(Brassicaceae)Withaniacoagulans(Solanaceae)ProvideessentialaminoacidsandsupportantioxidantdefensemechanismsinhumanmetabolismEasilydigestibleproteins(albumins)enhancenutrientabsorptionandimmunesupportDefense-relatedproteinsmayexhibitbioactiveandtherapeuticpropertiesinsmalldosesStorageproteinsincludepotentbioactivemolecules;requiredetoxificationforsafeuseHigh-qualityproteinscontributestocardiovascularhealthandmetabolicregulationRichinfunctionalproteins(e.g.,cruciferin)supportingdetoxificationandcellularprotectionBioactiveproteinsandpeptidesaidinimmunemodulationandglucoseregulationCucumismelovar.agrestis(Cucurbitaceae)NutritionallyimportantproteinscontributetohydrationbalanceandmetabolicsupportCucumismeloL.var.agrestisNaudin.NutritionallyimportantproteinscontributetohydrationbalanceandmetabolicsupportAntidiabetic,anti-inflammatory,antioxidantAntimicrobial,antifungal,hepatoprotectiveAnti-inflammatory,analgesicLaxative,antimicrobial,anti-inflammatoryCardioprotective,antioxidant,anticancerAntimicrobial,anti-inflammatorychemoprotectiveAntidiabetic,hepatoprotective,immunomodulatoryAntioxidant,anti-inflammatory,diureticAntioxidant,anti-inflammatory,diureticTable2showedthesemedicinalplantsareveryvaluableforhumannutritionandtherapeuticapplications,seedscontainfourtypesofprotein(SSPs)thatprovideabalanceofrapidlydigestibleproteins(albumins)andslowrelease,long-lastingproteins(globulins,prolamins,glutelins)tomeetimmediatemetabolicneeds,long-termaminoacidsupply,anddeliveryofbioactivecompounds,whicharedirectlylinkedtotheobservedantioxidant,antidiabetic,immunomodulatory,andcardioprotectiveactivitiesoftheseplants.L.usitatissimumL.,andL.inermisL.seedsarerichsourcesofalbuminandglobulinand,therefore,areespeciallyimportantforhumannutritionandtherapeuticuse.Fig.1.Graphicalrepresentationofthefourmajorseedstorageprotein(SSPs)(Albumin,Globulin,ProlaminandGlutelin.)concentrations(mg/g)measuredfromSyzygiumcuminiL.,LawsoniainermisL.Calotrpisprocera.RBroRicinuscommunisL.,LinumusitatissimumL.,BrassicajunceaLCzern.,WithaniacoagulansDun,andCucumismelovar.agrestisNaudin.seedsfoundinSindh,Pakistan.ConclusionThefindingsofthepresentstudyrevealthatthecompositionofseedstorageproteinsdiffersmarkedlyamongtheselectedmedicinalplantofSindh,Pakistan.AlbuminwasfoundtobethedominantproteinfractioninB.junceaLCzern,suggestingitskeyroleasaprimarywater-solublestorageproteininthisspecies.Contrastingly,R.communisLishighestamountofglobulin,suggestinganabundanceofsalt-solubleproteins.W.coagulansDun.hadauniqueproteinprofile,withprolaminbeingthedominantfraction,suggestinganadaptivemechanismofproteinstorage.S.cuminiLhadrelativelyhigherlevelsofglutelincomparedtotheotherspecies,butglutelinwasalwaystheleastabundantfraction.Thisindicatesthatseedstorageproteinprofilesarespecies-specificandinfluencedbygeneticandphysiologicalfactors,andalbumin,globulin,prolamin,andglutelinfractionsvaryinrelationtoseedstructure,nutritionalpotential,andmetabolicfunctions.G.2052Page1References1.Thangadurai,D.,(2001).NutritionalpotentialofbiochemicalcomponentsinGalactialongifoliaBenth.(Fabaceae).Nahrung/Food,45(2),97–100.2.Blagoveschenskii,A.V.(2006).Theevolutionofproteinsandtheevolutionoffloweringplants.Bio-chemistry(U.S.S.R.),25,9.3.Templeman,T.S.,A.E.Demaggio,andD.A.Stetler,(2012).Biochemistryoffernsporegermination:GlobulinstorageproteinsinMatteucciastruthiopterisL.PlantPhysiol.85,343-349.4.S.K.Khanzada,A.K.Khanzada,A.KandhroandA.Panhwar.(2021).Isolationandcharacterizationofmajorseedstorageproteins:II.ApiaceaefamilyfoundinSindh,Pakistan.PakistanJournalofScienceVol.73No.1,114-1195.Kumar,V.,etal.(2024).Influenceofseedpre-treatmentandstorageongerminationandphysiologicalcharacteristicsofmedicinalplantseeds.JournalofAppliedResearchonMedicinalandAromaticPlants,6.Latif,R.,&Nawaz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