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Purification and characterization of a novel antifungal protein secreted by Penicillium chrysogenum from an Arctic sediment

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Abstract

A fungal strain, Penicillium chrysogenum A096, was isolated from an Arctic sediment sample. Its culture supernatant inhibited mycelial growth of some plant pathogenic fungi. After saturation of P. chrysogenum A096 culture supernatant with ammonium sulfate and ion exchange chromatography, a novel antifungal protein (Pc-Arctin) was purified and identified by matrix assisted laser desorption ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF-TOF-MS). The gene encoding for Pc-Arctin consisting of 195 nucleotides was cloned from P. chrysogenum A096 to confirm the mass spectrometry result. Pc-Arctin displays antifungal activity against Paecilomyces variotii, Alternaria longipes, and Trichoderma viride at minimum inhibitory concentrations (MIC) of 24, 48, and 192 ng/disc, respectively. Pc-Arctin was most sensitive to proteinase K and then to trypsin but insensitive to papain. Pc-Arctin possesses high thermostability and cannot be antagonized by common surfactants, except for sodium dodecyl sulfate (SDS). Divalent ions, such as Mn2+, Mg2+, and Zn2+, inhibited the antifungal activity of Pc-Arctin. Hemagglutination assays showed that Pc-Arctin had no hemagglutinating or hemolytic activity against red blood cells (RBC) from rabbits, rats, and guinea pigs. Therefore, Pc-Arctin from Arctic P. chrysogenum may represent a novel antifungal protein with potential for application in controlling plant pathogenic fungal infection.

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Acknowledgments

We thank Xingzhong Qiao and Tianhua Zhong for their assistance with the experiments. The work was supported by grants from the Scientific Research Project of the Marine Public Welfare Industry of China (201005032, 201205020), the Nature Science Foundation of China (40930847), and China Polar Environment Comprehensive investigation and Assessment Program (CHINARE2013-01-06 and CHINARE2013-04-03).

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Authors and Affiliations

  1. State Key Lab. of Marine Environmental Science and Key Lab. of MOE for Coast and Wetland Ecosystems, School of Life Sciences, Xiamen University, Xiamen, China

    Zhiteng Chen, Wenchuan Yang & Tianling Zheng

  2. Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, No. 184, Daxue Road, Xiamen, 361005, China

    Zhiteng Chen, Jingqun Ao, Liping Jiao & Xinhua Chen

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  1. Zhiteng Chen
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  2. Jingqun Ao
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  3. Wenchuan Yang
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  4. Liping Jiao
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  5. Tianling Zheng
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  6. Xinhua Chen
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Correspondence to Tianling Zheng or Xinhua Chen.

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Chen, Z., Ao, J., Yang, W. et al. Purification and characterization of a novel antifungal protein secreted by Penicillium chrysogenum from an Arctic sediment. Appl Microbiol Biotechnol 97, 10381–10390 (2013). https://doi.org/10.1007/s00253-013-4800-6

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  • DOI: https://doi.org/10.1007/s00253-013-4800-6

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