Abstract
The hydroxyapatite/glycyrrhizin/lithium-based metal–organic framework (HA/GL/Li-MOF) nanocomposites were synthesized via the hydrothermal method in the presence of lecithin and glycyrrhizin. Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), and scanning electron microscopy (SEM) equipped with energy-dispersive X-ray spectroscopy (EDS) were applied for characterization of the fabricated nanocomposites. The HA/GL/Li-MOF and Li-MOF nanocomposites were employed as support for immobilization of Thermomyces lanuginosus lipase (TLL). The Plackett–Burman and Box-Behnken designs were used for screening and optimizing of variables affecting the immobilization conditions, respectively. The optimum specific activity of immobilized TLL on HA/GL/Li-MOF and Li-MOF nanocomposites (41.8 ± 1.2 U/mg and 39.4 ± 3.1 U/mg, respectively) was predictably determined at support concentration of 0.5 mg/mL, glutaraldehyde concentration of 5 mM, and enzyme activity of 20 U/mg, while the specific activities of TLL@ HA/GL/Li-MOF and TLL@Li-MOF were experimentally found to be 39.5 ± 3.7 U/mg and 38.5 ± 2.3 U/mg, respectively. The stability results showed that the TLL@ HA/GL/Li-MOF has suitable stability against pH and thermal denaturation. However, the immobilized TLL on Li-MOF represented lower stability compared with that of the HA/GL/Li-MOF. The immobilized TLL on HA/GL/Li-MOF maintained near 70% of its original activity after 15 days’ storage and during 5 runs of application. In addition, TLL@HA/GL/Li-MOF exhibited higher enzyme–substrate affinity (Km, 10.1 mM) compared to that of TLL@Li-MOF (Km, 23.4 mM). Therefore, these findings demonstrated the potential use of HA/GL/Li-MOF nanocomposites for enzyme immobilization.
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Research reported in this publication was supported by Elite Researcher Grant Committee under Award number 4000022 from the National Institute for Medical Research Development (NIMAD), Tehran, Iran. This work was financially supported by Pharmaceutics Research Center, Kerman University of Medical Sciences, Kerman, Iran (Grant number 98000501).
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Atefeh Ameri carried out the enzyme immobilization procedure, and its optimization as well as characterization of the immobilized enzyme and writing the manuscript. Fahimeh Asadi was involved in support preparation and screened the variables related to the immobilization process. Mojtaba Shakibaie was involved in designing of the experiments and analyzing of data. Alieh Ameri participated in analysis of the obtained results of support and immobilized enzyme characterization. Hamid Forootanfar supervised the study, analyzed the obtained results, and was involved in writing the manuscript. Mehdi Ranjbar was involved in support preparation and its characterization. All the authors read and approved the final manuscript.
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Ameri, A., Asadi, F., Shakibaie, M. et al. Hydroxyapatite/Glycyrrhizin/Lithium-Based Metal–Organic Framework (HA/GL/Li-MOF) Nanocomposite as Support for Immobilization of Thermomyces lanuginosus Lipase. Appl Biochem Biotechnol 194, 2108–2134 (2022). https://doi.org/10.1007/s12010-022-03800-3
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DOI: https://doi.org/10.1007/s12010-022-03800-3