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CLytA-DAAO Chimeric Enzyme Bound to Magnetic Nanoparticles. A New Therapeutical Approach for Cancer Patients?

ABSTRACT: D-amino acid oxidase (DAAO) is an enzyme that catalyzes the oxidation of D-amino acids generating H₂O₂. The enzymatic chimera formed by DAAO bound to the choline-binding domain of N-acetylmuramoyl-L-alanine amidase (CLytA) induces cytotoxicity in several pancreatic and colorectal carcinoma and glioblastoma cell models. In the current work, we determined whether the effect of CLytA-DAAO immobilized in magnetic nanoparticles, gold nanoparticles, and alginate capsules offered some advantages as compared to the free CLytA-DAAO. Results indicate that the immobilization of CLytA-DAAO in magnetic nanoparticles increases the stability of the enzyme, extending its time of action. Besides, we compared the effect induced by CLytA-DAAO with the direct addition of hydrogen peroxide, demonstrating that the progressive generation of reactive oxygen species by CLytA-DAAO is more effective in inducing cytotoxicity than the direct addition of H₂O₂. Furthermore, a pilot study has been initiated in biopsies obtained from pancreatic and colorectal carcinoma and glioblastoma patients to evaluate the expression of the main genes involved in resistance to CLytA-DAAO cytotoxicity. Based on our findings, we propose that CLytA-DAAO immobilized in magnetic nanoparticles could be effective in a high percentage of patients and, therefore, be used as an anti-cancer therapy for pancreatic and colorectal carcinoma and glioblastoma.

SUBMITTER: Fuentes-Baile M

PROVIDER: S-EPMC7867295 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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CLytA-DAAO Chimeric Enzyme Bound to Magnetic Nanoparticles. A New Therapeutical Approach for Cancer Patients?

Fuentes-Baile María M Pérez-Valenciano Elizabeth E García-Morales Pilar P de Juan Romero Camino C Bello-Gil Daniel D Barberá Víctor M VM Rodríguez-Lescure Álvaro Á Sanz Jesús M JM Alenda Cristina C Saceda Miguel M

International journal of molecular sciences 20210202 3

D-amino acid oxidase (DAAO) is an enzyme that catalyzes the oxidation of D-amino acids generating H<sub>2</sub>O<sub>2</sub>. The enzymatic chimera formed by DAAO bound to the choline-binding domain of N-acetylmuramoyl-L-alanine amidase (CLytA) induces cytotoxicity in several pancreatic and colorectal carcinoma and glioblastoma cell models. In the current work, we determined whether the effect of CLytA-DAAO immobilized in magnetic nanoparticles, gold nanoparticles, and alginate capsules offered ...[more]

PMID: 33540681

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Project description:Diabetes is a chronic metabolic disease which leads to high glucose levels in the blood, with severe consequences for human health. Due to the worldwide appeal for the reduction in calorie intake, this study presents the development of a nanomaterial able to capture sucrose selectively, thus providing a tool to remove naturally occurring sucrose from food, such as fruit juices, producing low-calorie juices for consumption. Magnetite nanoparticles (Fe3O4 NPs) coated with an inert material (SiO2) and functionalised with the enzyme invertase were designed to remove sucrose from solutions. Fe3O4 NPs were synthesised using the co-precipitation method, whereas the coating with a silica shell was done by the Stöber method. Its physicochemical characteristics were determined, with excellent stability over time. On the other hand, the invertase enzyme was extracted from dry Baker's yeast, purified and immobilised on the surface of the silica-coated Fe3O4 NPs. pH-triggered sucrose capture occurred at pH 3.0 once invertase with protonated catalytic residues was able just to bind with sucrose in a highly selective way. After a short, 1 min interaction, approximately 13.5 mmol L-1 of sucrose was captured per gram of nanomaterial and removed with the use of an external permanent magnet. The complex sucrose/nanomaterial was washed, and the released sucrose was put into buffered solution (pH = 4.8), where it underwent hydrolysis to yield inverted sugar. On the other side, sucrose-free nanomaterial was reused with no loss of enzymatic capability to capture sucrose at pH = 3.0 and maintained the invertase activity at pH 4.8 in ten consecutive rounds of re-use. As sucrose was recovered in the form of inverted sugar, not just low sugar beverage could be obtained, but also a high valued market product. Thus, the developed technology allows for the commercialisation of low-calorie food, offering healthier options to consumers and helping to fight diabetes and obesity.

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CLytA-DAAO Chimeric Enzyme Bound to Magnetic Nanoparticles. A New Therapeutical Approach for Cancer Patients?

Publications

CLytA-DAAO Chimeric Enzyme Bound to Magnetic Nanoparticles. A New Therapeutical Approach for Cancer Patients?

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