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Intracellular trehalose and sorbitol synergistically promoting cell viability of a biocontrol yeast, Pichia anomala, for aflatoxin reduction

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Abstract

Pichia anomala (Wickerhamomyces anomalus) WRL-076 was discovered by a visual screening bioassay for its antagonism against Aspergillus flavus. The yeast was shown to significantly inhibit aflatoxin production and the growth of A. flavus. P. anomala is a potential biocontrol agent for reduction of aflatoxin in the food chain. Maintaining the viability of biocontrol agents in formulated products is a great challenge for commercial applications. Four media, NYG, NYGS, NYGT and NYGST are described which support good growth of yeast cells and were tested as storage formulations. Post growth supplement of 5 % trehalose to NYGST resulted in 83 % viable yeast cells after 12 months in cold storage. Intracellular sorbitol and trehalose concentrations were determined by HPLC analysis at the beginning of the storage and at the end of 12 month. Correlation of cell viability to both trehalose and sorbitol suggested a synergistic effect. Bonferroni (Dunn) t Test, Tukey’s Studentized Range (HSD) Test and Duncan’s Multiple Range Test, all showed that yeast cell viability in samples with both intracellular trehalose and sorbitol were significantly higher than those with either or none, at a 95 % confidence level. DiBAC4(5) and CFDA-AM were used as the membrane integrity fluorescent stains to create a two-color vital staining scheme with red and green fluorescence, respectively. Yeast cells stored in formulations NYG and NYGS with no detectable trehalose, displayed mostly red fluorescence. Yeast cells in NYGST+5T showed mostly green fluorescence.

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Acknowledgments

This work is supported by in-house research program funds from U.S. Department of Agriculture, Agricultural Research Services (Project Numbers: 5325-42000-038, 5325-42000-048 and 5325-41440-006. We thank Thomas McKeon and Yuzhu Zhang for comments; Melissa Sanchez for technical assistance.

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. USDA-ARS, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA

    Sui Sheng T. Hua, Bradley J. Hernlem, Wallace Yokoyama & Siov Bouy L. Sarreal

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  1. Sui Sheng T. Hua
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  2. Bradley J. Hernlem
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  3. Wallace Yokoyama
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  4. Siov Bouy L. Sarreal
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Correspondence to Sui Sheng T. Hua.

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Hua, S.S.T., Hernlem, B.J., Yokoyama, W. et al. Intracellular trehalose and sorbitol synergistically promoting cell viability of a biocontrol yeast, Pichia anomala, for aflatoxin reduction. World J Microbiol Biotechnol 31, 729–734 (2015). https://doi.org/10.1007/s11274-015-1824-3

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  • DOI: https://doi.org/10.1007/s11274-015-1824-3

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