Laccase immobilized on cobalt ferrite magnetic nanoparticles for bioremoval of thymol blue dye

Abstract

Laccases are extensively studied by researchers because they can efficiently catalyze phenolic and non-phenolic compounds, as well as extremely recalcitrant environmental pollutants such as textile dyes. Between all carriers available for enzymatic immobilization, magnetic nanoparticles have the advantage of easy bioseparation by an external magnetic field from the reaction medium. Cobalt ferrite magnetic nanoparticles were synthesized and functionalized to immobilize laccases produced by Phlebia brevispora BAFC 633. The functionalization was made with 3-aminopropyltriethoxysilane followed by glutaraldehyde. The biocatalyst was employed to bioremove thymol blue from an aqueous solution. Magnetic nanoparticles were characterized by FTIR, TEM, DRX and magnetism, before and after immobilization. Immobilized laccases removed 86% of thymol blue in 23 h, and showed the same bioremoval capability even 3 cycles. The Michaelis-Menten constant was determinate, and immobilized laccases exhibit higher affinity to thymol blue and 2,6-dimethoxyphenol than free laccases. Regarding stability, immobilized laccases kept 78% of activity at 28 days while it was maintained in the refrigerator against free laccases, which reached 63% of initial activity at 28 days under the same conditions. The effective concentration 50 from Lactuca sativa was near 100% when the effluent was diluted at least 1:1 in water. Thus, the biocatalyst obtained has a potential approach in the bioremoval and detoxification of dyes such as thymol blue.

Keywords

Nanobioremediation / Thymol blue / Immobilization / Magnetic nanoparticles / Laccase / Chemical Sciences / Physical Chemistry (incl. Structural)

References

[1] Al-MaqdiKA, ElmerhiN, AlzamlyA, ShahI, AshrafSS. Laccase–copper phosphate hybrid nanoflower as potent thiazole remediation agent. J Water Process Eng, 2023.

[2] EcerÜ, YılmazŞ. Fabrication of magnetic biochar-MIL-68(Fe)-supported cobalt composite material toward the catalytic reduction performance of crystal violet. J Water Process Eng, 2024.

[3] LeontieșAR, RăducanA, CulițăDC, AlexandrescuE, MoroșanA, MihaiescuDE, AricovL. Laccase immobilized on chitosan-polyacrylic acid microspheres as highly efficient biocatalyst for naphthol green B and indigo carmine degradation. Chem Eng J, 2022.

[4] KatiyarJ, SaharanVK. Enhanced photocatalytic degradation of reactive blue 21 dye and textile dyeing effluent by synthesized SmFeO₃-rGO photocatalyst in combination with ultrasonication: characterization and performance evaluation. J Water Process Eng, 2023.

[5] GeorgeJ, RajendranDS, VenkataramanS, RathankumarAK, SaikiaK, MuthusamyS, SinghI, SinghI, SinhaS, RamkumarS, CabanaH, VaidyanathanVK. Insolubilization of Tramates Versicolor laccase as cross-linked enzyme aggregates for the remediation of trace organic contaminants from municipal wastewater. Environ Res, 2022.

[6] BilalM, RashidEU, ZdartaJ, dos SantosJCS, FernandesPCB, ChengH, JesionowskiT. Engineering magnetic nanobiocatalytic systems with multipurpose functionalities for biocatalysis, biotechnology and bioprocess applications. Sustain Chem Pharm, 2022.

[7] Cardoso PintoG, LucenaGN, Debone PiazzaR, Lopes CostaJM, Torres Couto Coimbra e SilvaE, GuY, Veloso de PaulaA, JoaoN, SilvaO, Costa MarquesRF. Evaluation of the alternating magnetic field (AMF) influence in catalytic activities of enzymes immobilized into magnetic graphene oxide: a new approach. Mater Today Commun, 2023.

[8] MondalK, SharmaP, NajarIN, MondalS, HalderSK, ThakurN, MondalKC. Cloning, expression, and characterization of thermostable α-amylase of a thermophilic bacterium isolated from hot-spring of Sikkim and sustainable immobilization on coconut coir. Syst Microbiol Biomanufacturing, 2024.

[9] Naghash-HamedS, ArsalaniN, MousaviSB. Facile copper ferrite/carbon quantum dot magnetic nanocomposite as an effective nanocatalyst for reduction of para-nitroaniline and Ortho-Nitroaniline. Nano Futur, 2022.

[10] ChatterjeeA, MridhaD, BanerjeeJ, ChandaS, RayK, AcharyaK, DasM, RoychowdhuryT, SarkarJ. Green synthesis of iron oxide nanoparticles and their ameliorative effect on arsenic stress relief in Oryza sativa seedlings. Biocatal Agric Biotechnol, 2021.

[11] WuK, LiuJ, ChughVK, LiangS, SahaR, KrishnaVD, CheeranMJC, WangJP. Magnetic nanoparticles and magnetic particle spectroscopy-based bioassays: a 15 year recap. Nano Futur, 2022.

[12] HunyekA, SirisathitkulC. Effect of cobalt ferrite on curing and electromagnetic properties of natural rubber composites. Adv Mater Res, 2023.

[13] Vinod Kumar, SinghD, SinghB. Mycogenic synthesis of iron nanoparticles using thermophilic mould Myceliophthora Thermophila and their applicability in environmental remediation. Biocatal Agric Biotechnol, 2024.

[14] Prigioni GA, Rodríguez MD, Zapata PD, Villalba LL. Licor De maíz como medio de cultivo para la producción de lacasa por phlebia brevispora. Rev Cienc Y Tecnol. 2018;20(30):1–12.

[15] FonsecaMI, FariñaJI, CastrilloML, RodríguezMD, NuñezCE, VillalbaLL, ZapataPD. Biopulping of wood chips with Phlebia Brevispora BAFC 633 reduces lignin content and improves pulp quality. Int Biodeterior Biodegrad, 2014.

[16] FonsecaMI, TejerinaMR, Sawostjanik-AfanasiukSS, GiorgioEM, BarchukML, ZapataPD, VillalbaLL. Preliminary studies of new strains of Trametes sp. From Argentina for laccase production ability. Brazilian J Microbiol, 2016.

[17] RanF, ZouY, XuY, LiuX, ZhangH. Fe₃O₄@MoS₂@PEI-facilitated enzyme tethering for efficient removal of persistent organic pollutants in water. Chem Eng J, 2019.

[18] YoungBJ, RieraNI, BeilyME, BresPA, CrespoDC, RoncoAE. Toxicity of the effluent from an anaerobic bioreactor treating cereal residues on Lactuca sativa. Ecotoxicol Environ Saf, 2012.

[19] CruzYWG, VieiraYA, VilarDS, TorresNH, AguiarMM, CavalcantiEB, Américo-PinheiroJHP, SorianoRN, BharagavaRN, LimaAS, FerreiraLFR. Pulp wash: a new source for production of ligninolytic enzymes and biomass and its toxicological evaluation after biological treatment. Environ Technol, 2020.

[20] QiuX, WangY, XueY, LiW, HuY. Laccase immobilized on magnetic nanoparticles modi fi ed by amino- functionalized ionic liquid via dialdehyde starch for phenolic compounds biodegradation. Chem Eng J, 2020.

[21] HosseiniSH, DizMM. Preparation and characterization of nanoflake composite multi core–shell SrFe₁₂O₁₉/Fe₃O₄/PEG/PPy. Adv Mater Res, 2012.

[22] da SilvaPMM, CamparottoNG, Figueiredo NevesT, MastelaroVR, NunesB, Franco PiconeCS, PredigerP. Instantaneous adsorption and synergic effect in simultaneous removal of complex dyes through nanocellulose/graphene oxide nanocomposites: batch, fixed-bed experiments and mechanism. Environ Nanatechnol Monit Manag, 2021.

[23] WanJ, ZhangL, YangB, JiaB, YangJ, SuX. Enzyme immobilization on amino-functionalized Fe₃O₄@SiO₂ via electrostatic interaction with enhancing biocatalysis in sludge dewatering. Chem Eng J, 2022.

[24] ZhangM, JiangS, GuoX, TangX, BaiL, WangJ, ZhangH, XuD, WuR, LiuL, LiangH. Fabrication of high permeability and antifouling composite membrane loaded with Fe₃O₄ nanoparticles via a magnetic field induced phase separation. J Water Process Eng, 2023.

[25] ZhangF, WangL, YangL, DaiY, ZhangJ, HeL, CuiJ, ShenJ, WangZ. Recoverable magnetic Fe-MOF immobilized carrier to immobilize two microorganisms for reduction of heavy oil viscosity and oilfield wastewater COD. J Water Process Eng, 2023.

[26] WilliamsonGK, HallWH. X-ray line broadening from filed aluminium and wolfram. Acta Metall, 1953.

[27] OhY, SahuM, HajraS, PadhanAM, PandaSKimHJ. Spinel ferrites (CoFe₂O₄): synthesis, magnetic properties, and electromagnetic generator for vibration energy harvesting. J Electron Mater, 2022.

[28] ErsenO, BéginS, HoulléM, AmadouJ, JanowskaI, GrenècheJM, CrucifixC, Pham-HuuC. Microstructural investigation of magnetic CoFe2O4 nanowires inside carbon nanotubes by electron tomography. Nano Lett, 2008.

[29] FonsecaMI, MolinaMA, WinnikD, BusiMV, FarinaJI, VillalbaLL, ZapataPD. Isolation of a laccase-coding gene from the lignin-degrading fungus Phlebia Brevispora BAFC 633 and heterologous expression in Pichia pastoris. J Appl Microbiol, 2018.

[30] Syukri D, Suryanto H, Kurniawan F, Hari PD, Fiana RM, Rini. Bacterial reduction in river water using nanocellulose membrane from pineapple biomass with ferrous-ferric oxide reinforcement. Glob J Environ Sci Manag. 2024. https://doi.org/10.22035/gjesm.2024.02.14.

[31] IllanesA, Fernandez-LafuenteR, GuisánJM, WilsonLIllanesA. Heterogeneous enzyme kinetics. Enzyme Biocatalysis. Principles and applications, 2008ChileSpringer155-203.

[32] SheldonRA. Enzyme immobilization: the Quest for Optimum Performance. Adv Synth Catal, 2007.

[33] BarberisS, GuzmanF, IllanesA, Lopez-SantínJIllanesA. Study cases of enzymatic processes. Enzyme Biocatalysis. Principles and applications, 2008ChileSpringer253-378.

[34] Akkuş ÇetinusŞ, ÖztopHN. Immobilization of catalase on chitosan film. Enzyme Microb Technol, 2020.

[35] DwyerDF, KrummeML, BoydSA, TiedjeJM. Kinetics of phenol biodegradation by an immobilized methanogenic consortium. Appl Environ Microbiol, 1986.

[36] HuangQ, ShindoH. Effects of copper on the activity and kinetics of free and immobilized acid phosphatase. Soil Biol Biochem, 2000.

[37] ZhangH, HayAG. Magnetic biochar derived from biosolids via hydrothermal carbonization: enzyme immobilization, immobilized-enzyme kinetics, environmental toxicity. J Hazard Mater, 2020.

[38] NadarogluH, MosberG, GungorAA, AdıguzeldG, AdiguzelA. Biodegradation of some azo dyes from wastewater with laccase from Weissella viridescens LB37 immobilized on magnetic chitosan nanoparticles. J Water Process Eng, 2019.

[39] LiuY, ZengZ, ZengG, TangL, PangY, LiZ, LiuC, LeiX, WuM, RenP, LiuZ, ChenM, XieG. Immobilization of laccase on magnetic bimodal mesoporous carbon and the application in the removal of phenolic compounds. Bioresour Technol, 2012.

[40] KadamAA, JangJ, LeeDS. Supermagnetically tuned halloysite nanotubes functionalized with aminosilane for covalent laccase immobilization. ACS Appl Mater Interfaces, 2017.

[41] Ayala SchimpfAR, OrtelladoLE, GamarraMD, FonsecaMI, ZapataPD. In Vitro and computational response of Differential Catalysis by Phlebia Brevispora BAFC 633 Laccase in Interaction with 2,4-D and Chlorpyrifos. Int J Mol Sci, 2024.

Funding

Agencia Nacional de Promoción Científica y Tecnológica(PICT-2020-03105)

RIGHTS & PERMISSIONS

Jiangnan University