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A Possible Role of Aspergillus niger Mitochondrial Cytochrome c in Malachite Green Reduction Under Calcium Chloride Stress

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Abstract

In previous work, decolorization of malachite green (MG) was studied in Aspergillus niger in the presence and absence of calcium chloride stress. Decolorization took place within 24 h, and a signal transduction process that initiated MG decolorization was suggested to be involved. In the present study, further investigation of the relationship between calcium chloride stress and enhanced MG biodegradation was conducted at the sub-cellular level. MG-NADH reductase activity, a key enzyme in MG decolorization, was produced as decolorization commenced, and enzyme activity increased threefold upon exposure to calcium chloride. Inhibitors of cytochrome p450, Ca2+ channel activity as well as activity of the signaling protein phosphoinositide 3-kinase were tested. All three activities were inhibited to different extents resulting in reduced MG decolorization. Spectral analysis of the mitochondrial fraction showed a heme signal at 405 nm and A405/A280 ratio that is characteristic of the porphoryin ring of cytochromes. There were no peaks detected for cytochromes a or b, but a shoulder appearing at 550 nm was observed, which suggested that cytochrome c is involved; the absorbance for cytochrome c doubled after calcium chloride stress supporting this idea. MG decolorization took place via a series of demethylation steps, and cytotoxicity analysis revealed a decrease in the toxicity associated with generation of leucomalachite green.

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Acknowledgments

This work was supported by US–Egypt joint Grant, Project ID: 750, Science and Technology Development Fund (STDF), Cairo, Egypt. The authors would also like to thank the Cancer Research Laboratory at the road to Nobel, National Research Center, Cairo, Egypt, for providing their laboratory facilities for the cytotoxicity tests.

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Authors and Affiliations

  1. Microbiology Department, National Center for Radiation Research and Technology (NCRRT), Atomic Energy Authority, 3 Ahmad El Zomor St., Nasr City, Cairo, 11371, Egypt

    Ola M. Gomaa

  2. Radiation Physics Department, National Center for Radiation Research and Technology (NCRRT), Atomic Energy Authority, 3 Ahmad El Zomor St., Nasr City, Cairo, 11371, Egypt

    Nabila S. Selim

  3. Department of Food Science and Human Nutrition, Michigan State University (MSU), East Lansing, MI, USA

    John E. Linz

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  1. Ola M. Gomaa
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  2. Nabila S. Selim
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  3. John E. Linz
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Correspondence to Ola M. Gomaa.

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Gomaa, O.M., Selim, N.S. & Linz, J.E. A Possible Role of Aspergillus niger Mitochondrial Cytochrome c in Malachite Green Reduction Under Calcium Chloride Stress. Cell Biochem Biophys 67, 1291–1299 (2013). https://doi.org/10.1007/s12013-013-9661-1

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  • DOI: https://doi.org/10.1007/s12013-013-9661-1

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