Hydrochar as protein support: preservation of biomolecule properties with non-covalent immobilization
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In this work, the ConBr lectin was non-covalently immobilized onto hydrochar (HC). This carbonaceous material was produced by the hydrothermal carbonization of glucose and then put to interact with the lectin, aiming to immobilize the biomolecule via electrostatic interactions. Samples obtained after the interaction were characterized by CHNS elemental analysis, scanning electron microscopy and Fourier transform infrared spectroscopy (FTIR). FTIR results from the conjugated sample identified the presence of NH2 + and NH3 + groups of the protein and COO− groups of the HC, indicating the occurrence of electrostatic interaction between the biomolecule and the support. Furthermore, the immobilization experiment was also performed using ConBr lectin marked with fluorescein isothiocyanate to assess the immobilization on the hydrochar using fluorescence emission analysis. Hemagglutination tests revealed that even after the conjugation with the HC, the agglutinating property of lectin toward erythrocytes (red blood cells) was preserved. Finally, our results indicate that non-covalent interactions represent an efficient mechanism for protein immobilization on the HC while maintaining the protein structure and its biological activity.
The authors acknowledge Francisco Holanda Soares Júnior for the SEM images and Antonio Gomes de Souza Filho for the precious discussions. Also, the authors are grateful to Central Analítida-UFC/CT-INFRA/MCTI-SISNANO/Pró-Equipamentos CAPES for providing the electron scanning microscopes and confocal fluorescence microscope and CETENE for the TEM measurements. This work was supported by CNPq (Grant 478743/2013-0), FUNCAP (PRONEX PR2-0101-00006.01.00/15) and CAPES (for the scholarship awarded to M. O. Castro).
- 2.Benešová E, Králová B (2012) Affinity interactions as a tool for protein immobilization. In: Magdeldin S (ed) Affinity chromatography. InTech, Rijeka, pp 29–46Google Scholar
- 14.Cavada BS, Barbosa T, Arruda S et al (2001) Revisiting proteus: do minor changes in lectin structure matter in biological activity? Lessons from and potential biotechnological uses of the Diocleinae subtribe lectins. Curr Protein Pept Sci 2:123–135. doi: 10.2174/1389203013381152 CrossRefGoogle Scholar
- 34.Pavia DL, Lampman GM, Kriz GS (2001) Introduction to spectroscopy: a guide for students of organic chemistry, 3rd edn. Thomson Learning Inc, StamfordGoogle Scholar
- 35.Patnaik P (2004) Dean’s analytical chemistry book, 2nd edn. McGRAW-HILL, New YorkGoogle Scholar
- 36.Silverstein RM, Webster FX, Kiemle DJ (2006) Identificação Espectrométrica de Compostos Orgânicos, 7th edn. LTC, Rio de JaneiroGoogle Scholar
- 39.Portal, ExPASy—Bioinformatics Resource (ProtParam Tool). http://web.expasy.org/cgi-bin/protparam/protparam1?P55915@1-237@. Accessed 8 Sep 2016