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Activity of chitosan–lysozyme nanoparticles on the growth, membrane integrity, and β-1,3-glucanase production by Aspergillus parasiticus

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Abstract

Synthesis of nanocomposites from antimicrobial biopolymers such as chitosan (CS) and lysozyme (LZ) is an important and promising area in bionanotechnology. Chitosan–lysozyme (CS–LZ) nanoparticles (NPs) were prepared by the nanoprecipitation method, using commercial chitosan of 153 kDa. TEM and dynamic light scattering (DLS) analysis were carried out to evaluate the morphology, size, dispersion, and Z potential. Association efficiency of lysozyme was determined using Coomassie blue assay. The antifungal activity of NPs against Aspergillus parasiticus was evaluated through cell viability (XTT), germination and morphometry of spores, and reducing sugars production; the effects on membrane integrity and cell wall were also analyzed. NPs’ size were found in the range of 13.4 and 11.8 nm for CS–LZ and CS NPs, respectively, and high Z potential value was observed in both NPs. Also, high association of lysozyme was presented in the CS matrix. With respect to the biological responses, CS–LZ NPs reduced the viability of A. parasiticus and a strong inhibitory effect on the germination of spores (100% of inhibition) was observed at 24 h in in vitro assays. CS–LZ and CS NPs affected the membrane integrity and the cell wall of spores of fungi with respect to control, which is consistent with the low amount of reducing sugars detected. CS–LZ NPs prepared by nanoprecipitation promise to be a viable and safe alternative for use in biological systems, with a possible low or null impact to humans and biota. However, the potential benefits and the environmental and health implications of NPs need to be globally discussed due to its possible negative effects.

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Abbreviations

CS–LZ:

Chitosan–lysozyme

NPs:

Nanoparticles

CS:

Chitosan

LZ:

Lysozyme

TEM:

Transmission electron microscopy

DLS:

Dynamic light scattering

XTT:

Tetrazolium salt

Mw:

Molecular weight

ROS:

Reactive oxygen species

kDa:

Kilo Daltons

PDA:

Potato dextrose agar

µg/mL:

Microgram per milliliter

BSA:

Bovine serum albumin

PI:

Propidium ioidide

CW:

Calcofluor-white

CS:

Chitosan of 153 kDa

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Acknowledgements

The study was funded by the Mexican Council for Science and Technology (CONACyT) through the project No. 219786 (CB-2013-01) and for the scholarship to Cynthia Nazareth Hernández-Téllez for postgraduate studies. The authors acknowledge the University of Sonora for the assistantship grant to Francisco Julián Rodríguez-Córdova for undergraduate studies.

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Authors and Affiliations

  1. Laboratorio de Microbiología y Micotoxinas, Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Blvd. Luis Encinas y Rosales s/n, Col. Centro, 83000, Hermosillo, Sonora, Mexico

    Cynthia Nazareth Hernández-Téllez, Francisco Julián Rodríguez-Córdova, Ema Carina Rosas-Burgos, Mario Onofre Cortez-Rocha, Armando Burgos-Hernández, Wilfrido Torres-Arreola, Aarón Martínez-Higuera & Maribel Plascencia-Jatomea

  2. Laboratorio de Biopolímeros, Centro de Investigación en Alimentación y Desarrollo, A.C, Hermosillo, Sonora, Mexico

    Jaime Lizardi-Mendoza

Authors
  1. Cynthia Nazareth Hernández-Téllez
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  2. Francisco Julián Rodríguez-Córdova
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  3. Ema Carina Rosas-Burgos
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  4. Mario Onofre Cortez-Rocha
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  6. Jaime Lizardi-Mendoza
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  7. Wilfrido Torres-Arreola
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  9. Maribel Plascencia-Jatomea
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Contributions

Maribel Plascencia-Jatomea conceived and designed the experiments. Cynthia Nazareth Hernández-Téllez, Francisco Julián Rodríguez-Córdova and Aaron Martínez-Higuera performed the experiments. Ema Carina Rosas Burgos, Mario Onofre Cortez-Rocha, Armando Burgos-Hernández, Wilfrido Torres-Arreola and Jaime Lizardi-Mendoza analyzed the data and contributed with reagents, materials and analysis tools. Cynthia Nazareth Hernández-Téllez and Maribel Plascencia-Jatomea wrote the paper.

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Correspondence to Maribel Plascencia-Jatomea.

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Hernández-Téllez, C.N., Rodríguez-Córdova, F.J., Rosas-Burgos, E.C. et al. Activity of chitosan–lysozyme nanoparticles on the growth, membrane integrity, and β-1,3-glucanase production by Aspergillus parasiticus . 3 Biotech 7, 279 (2017). https://doi.org/10.1007/s13205-017-0913-4

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  • DOI: https://doi.org/10.1007/s13205-017-0913-4

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