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Structure and enzymatic activity of laser immobilized ribonuclease A

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Abstract

Ribonuclease A (RNase A) enzyme was immobilized on solid holders by matrix-assisted pulsed laser evaporation (MAPLE) technique. The experiments were performed inside a stainless steel irradiation chamber. A UV KrF* (λ = 248 nm, τFWHM ≅ 25 ns, ν = 10 Hz) excimer laser source was used for irradiations. Surface morphology, molecular structure, and enzymatic activity of laser transferred RNase A samples were investigated as a function of RNase A concentration in the frozen composite MAPLE targets. Surface morphology and thickness of the immobilized enzyme were investigated by atomic force microscopy, scanning electron microscopy, and surface profilometry. The molecular structure of the laser transferred RNase A was determined by Fourier transform infrared spectroscopy. The enzymatic activity of RNase A after immobilization was tested through ribonucleic acid removal from deoxyribonucleic acid (DNA) extract solutions isolated from plant and animal tissues. A molecular method based on polymerase chain reaction was used to investigate the functional properties of DNA extracts treated with laser immobilized RNase A.

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Acknowledgements

The authors acknowledge with thanks the financial support of the Executive Unit for Financing Higher Education, Research, Development, and Innovation of the Romanian Ministry of Education, Research, Youth, and Sports under the Grant PN-II-RU-PD-2011-3-0241 and PN-II-PT-PCCA-2011-3.2-1235.

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Authors and Affiliations

  1. National Institute for Lasers, Plasma and Radiations Physics, P. O. Box MG 36, 77125, Bucharest, Romania

    C. Popescu, A. C. Popescu, I. Iordache, M. Motoc & E. Gyorgy

  2. Department of Genetics, Faculty of Biology, University of Bucharest, 060101, Bucharest, Romania

    D. Pojoga, A. Simon-Gruita, N. Constantin & G. Duta Cornescu

  3. Consejo Superior de Investigaciones Científicas, Instituto de Ciencia de Materiales de Barcelona (CSIC-ICMAB), Campus UAB, 08193, Bellaterra, Spain

    E. Gyorgy

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  1. C. Popescu
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Correspondence to E. Gyorgy.

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Popescu, C., Popescu, A.C., Iordache, I. et al. Structure and enzymatic activity of laser immobilized ribonuclease A. J Mater Sci 49, 4371–4378 (2014). https://doi.org/10.1007/s10853-014-8136-0

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  • DOI: https://doi.org/10.1007/s10853-014-8136-0

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