Abstract
For the first time, 3D chitin scaffolds from the marine demosponge Aplysina archeri were used for adsorption and immobilization of laccase from Trametes versicolor. The resulting chitin–enzyme biocatalytic systems were applied in the removal of tetracycline. Effective enzyme immobilization was confirmed by scanning electron microscopy. Immobilization yield and kinetic parameters were investigated in detail, in addition to the activity of the enzyme after immobilization. The designed systems were further used for the removal of tetracycline under various process conditions. Optimum process conditions, enabling total removal of tetracycline from solutions at concentrations up to 1 mg/L, were found to be pH 5, temperature between 25 and 35 °C, and 1 h process duration. Due to the protective effect of the chitinous scaffolds and stabilization of the enzyme by multipoint attachment, the storage stability and thermal stability of the immobilized biomolecules were significantly improved as compared to the free enzyme. The produced biocatalytic systems also exhibited good reusability, as after 10 repeated uses they removed over 90% of tetracycline from solution. Finally, the immobilized laccase was used in a packed bed reactor for continuous removal of tetracycline, and enabled the removal of over 80% of the antibiotic after 24 h of continuous use.
Highlights
Diverse polymer, carbon, inorganic, and hybrid-based ultralight porous three-dimensional (3D) materials have been used for enzyme immobilization [1,2,3,4]
For the first time, 3D chitin scaffolds from the marine demosponge A. archeri were used as a support for effective immobilization of laccase
The thermal and storage stability of the immobilized enzyme were significantly improved as compared to the free enzyme, indicating the protective effect of the support on the biomolecules
Summary
Diverse polymer-, carbon-, inorganic-, and hybrid-based ultralight porous three-dimensional (3D) materials have been used for enzyme immobilization [1,2,3,4]. Modern porous 3D printing scaffolds with complex internal structures and channels have been the focus of much attention [5]. Our scaffolding strategy [6] is based, on naturally prefabricated 3D scaffolds of poriferan (sponge) origin. These unique biopolymer-containing constructs offer alternative immobilization matrices that can be isolated from demosponges cultivated worldwide, to provide appropriate supports for a broad range of enzymes. SbpDpeexoeritoncnrttgeieromeiencspee)aorc[are7btto,ie8iduod]asmraesi(smppalroeseenoftsfigecekinscnnttiovsa[cw9ea–rnfe1ifnn2oae]l,asdwpscawpowblamlihecsamticsetioheournthcrrsicaeaevalientombfraeeetpxnchtretronesttmpe[l1iyone3nab–gbc1eeei7eoson])m,ur[si7emesp,l8epeo]ctorittncrreeosgdpci[hnr9aee–sssm1ece2naifs]fft,tferocawyltdiarvses[en1twe8ein]hwt,iracaehpbaapnltehmdliacsevaoneetutinorrz[cen1yecs3me–oinn1eft7ly], eliemcmtroocbhileimzaitsitorny [[1198]].,Manedanewnzhyilme,emimarmineobdielimzaotsiopnon[1g9e]s. oMf tehaenowrhdieler,Vmeraorningeiiddeamhaovspe obneegnesreocfotghneizoerdder Vaesroanrgeiindeawahbalveesobueercne roefcuognniqizueedly apsrea-strreuncetwuraebdle3Dsocuhricteinoofususnciaqfufoelldysp[2re0-–s2tr8u]cwtuhriecdh h3Davechfoituinnodus scaapfpfolilcdasti[o2n0–s2in8]tiwsshuicehenhgaivneefeoruinngd[6a,p2p1,l2ic9a–t3io4n],sdirnutgisrseuleeaesneg[i3n5e],etrhinegd[e6v,2e1lo,2p9m–3e4n]t, odfruhgybrreildeamseat[e3r5i]a,ltshe d[e3v6e–l4o0p]m, aenndt oefnhvyirbornidmmenattaelrisaclisen[3c6e–[4401],,4a2n].dCehnivtiinroonfminevnetartlesbcrieanteceor[4ig1i,n42h].asCphriteivnioofuisnlyvebreteebnrsattuedoireidgin hbays prreesveiaorucshleyrbseienn pstruodteieind bimymreosebailriczhaetriosninapsroateminaitmrixmfoobrilitzhaetiiomnmasoabimlizaatrtiixonforoftheenizmymmoebsil[i4z3a–ti4o8n],of eninzcylumdeisng[43li–p4a8s]e,sin[c4l9u,5d0in] ganlidpapsaeps a[4in9,5[501]]a. nHdopwaepvaeinr, [i5n1]t.hHisoswtuedvyer,wine tihnivsessttuigdaytewdethinevaebstiligitayteodf tahe aubinliitqyuoe,f raeuadnyiq-tuoe-,urseead3Dy-tcoh-uitsineo3uDs cshcaitfifnooldusissoclaaftfeodldfriosomlattehde fmroamrintehedmemaroisnpeodnegme oAspplyosningae Aarpclhyesriina ar(cFhigerui r(eFi1g)utroe i1m) tmo oimbimlizoebillaiczcealsaeccaassea asselaecsteeldecetendzyenmzey,mane,daannd iannnionvnaotvivaetivapepalpicpalitcioantioonf othf itshiusnuiqnuiqeue sysystsetmemininththeerermemovoavlaol fotfettertarcayccylcinlienbeybysimsimulutaltnaenoeuosuasdasdosroprtpiotnioannadncdatcaaltyatliyctciconcovnervseirosnio. n.
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