Improved L-Asparaginase Properties and Reusability by Immobilization onto Functionalized Carbon Xerogels

resumo

Enzyme immobilization can offer a range of significant advantages, including reusability, and increased selectivity, stability, and activity. In this work, a central composite design (CCD) of experiments and response surface methodology (RSM) were used to study, for the first time, the L-asparaginase (ASNase) immobilization onto functionalized carbon xerogels (CXs). The best results were achieved using CXs obtained by hydrothermal oxidation with nitric acid and subsequent heat treatment in a nitrogen flow at 600 degrees C (CX-OX-600). Under the optimal conditions (81 min of contact time, pH 6.2 and 0.36 g/L of ASNase), an immobilization yield (IY) of 100 % and relative recovered activity (RRA) of 103 % were achieved. The kinetic parameters obtained also indicate a 1.25-fold increase in the affinity of ASNase towards the substrate after immobilization. Moreover, the immobilized enzyme retained 97 % of its initial activity after 6 consecutive reaction cycles. All these outcomes confirm the promising properties of functionalized CXs as support for ASNase, bringing new insights into the development of an efficient and stable immobilization platform for use in the pharmaceutical industry, food industry, and biosensors. The development of an efficient strategy for immobilizing L-asparaginase enzyme onto functionalized carbon xerogels is the focus of this work. The results reveal the importance of tuning the surface chemistry of the materials and prove the enhanced activity, higher affinity with the substrate, and reusability of L-asparaginase achieved through immobilization.+ image

palavras-chave

ACUTE LYMPHOBLASTIC-LEUKEMIA; SURFACE-CHEMISTRY; ACTIVATED CARBON; ADSORPTION; LACCASE; EXPRESSION; STABILITY; NANOTUBES; CHILDREN; ANTIBODY

categoria

Chemistry

autores

Barros, RAM; Cristovao, RO; Carneiro, IG; Barros, MA; Pereira, MM; Carabineiro, SAC; Freire, MG; Faria, JL; Santos-Ebinuma, VC; Tavares, APM; Silva, CG

Grupos

G5 - Materiais Biomiméticos, Biológicos e Vivos
G6 - Materiais Virtuais e Inteligência Artificial

Projectos

CICECO - Aveiro Institute of Materials (UIDB/50011/2020)

CICECO - Aveiro Institute of Materials (UIDP/50011/2020)

Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)

Collaboratory for Emerging Technologies, CoLab (EMERGING TECHNOLOGIES)

agradecimentos

This work was supported by national funds through FCT/MCTES (PIDDAC): LSRE-LCM, UIDB/50020/2020 (DOI: 10.54499/UIDB/50020/2020) and UIDP/50020/2020 (DOI: 10.54499/UIDP/50020/2020); and ALiCE, LA/P/0045/2020 (DOI: 10.54499/LA/P/0045/2020). This work was developed partly within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020 (DOI 10.54499/UIDB/50011/2020), UIDP/50011/2020 (DOI 10.54499/UIDP/50011/2020) & LA/P/0006/2020 (DOI 10.54499/LA/P/0006/2020), financed by national funds through the FCT/MCTES (PIDDAC). This work received support from Portuguese national funds through projects DOIs: 10.54499/LA/P/0008/2020, 10.54499/UIDP/50006/2020, 10.54499/UIDB/50006/2020. Valeria C. Santos-Ebinuma acknowledges FAPESP (2018/06908-8 and 2021/06686-8) and the National Council of Scientific and Technological Development, Brazil (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico - CNPq) - Grant no. 312463/2021-9. A.P.M.T. acknowledges FCT for the research contract CEEC-IND/2020/01867 (doi: 10.54499/2020.01867.CEECIND/CP1589/CT0013). S.A.C.C. also acknowledges FCT for the Scientific Employment Stimulus - Institutional Call (DOI 10.54499/CEECINST/00102/2018/CP1567/CT0026). M. M. P. acknowledges the financial support of FCT, Portugal, within the projects DOI: 10.54499/UIDB/00102/2020 (Base funding) and DOI: 10.54499/UIDP/00102/2020 (Programmatic funding). M.A.B. and R.A.M.B. acknowledge FCT for their PhD grants SFRH/BD/145014/2019 and 2022.12055.BD, respectively. I.G.C. acknowledges her scholarship funded by Project #SUMMER@LSRE-LCM_2021, program "Verao com Ciencia", financed by national funds through FCT/MCTES (PIDDAC).