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. 2021 Apr 21;13(5):294.
doi: 10.3390/toxins13050294.

A Novel Microbial Zearalenone Transformation through Phosphorylation

Yan Zhu  1 , Pascal Drouin  2 , Dion Lepp  1 , Xiu-Zhen Li  1 , Honghui Zhu  1 , Mathieu Castex  2 , Ting Zhou  1
Affiliations

A Novel Microbial Zearalenone Transformation through Phosphorylation

Yan Zhu et al. Toxins (Basel). .

Abstract

Zearalenone (ZEA) is a mycotoxin widely occurring in many agricultural commodities. In this study, a purified bacterial isolate, Bacillus sp. S62-W, obtained from one of 104 corn silage samples from various silos located in the United States, exhibited activity to transform the mycotoxin ZEA. A novel microbial transformation product, ZEA-14-phosphate, was detected, purified, and identified by HPLC, LC-MS, and NMR analyses. The isolate has been identified as belonging to the genus Bacillus according to phylogenetic analysis of the 16S rRNA gene and whole genome alignments. The isolate showed high efficacy in transforming ZEA to ZEA-14-phosphate (100% transformation within 24 h) and possessed advantages of acid tolerance (work at pH = 4.0), working under a broad range of temperatures (22-42 °C), and a capability of transforming ZEA at high concentrations (up to 200 µg/mL). In addition, 23 Bacillus strains of various species were tested for their ZEA phosphorylation activity. Thirteen of the Bacillus strains showed phosphorylation functionality at an efficacy of between 20.3% and 99.4% after 24 h incubation, suggesting the metabolism pathway is widely conserved in Bacillus spp. This study established a new transformation system for potential application of controlling ZEA although the metabolism and toxicity of ZEA-14-phosphate requires further investigation.

Keywords: Bacillus; microbial transformation; phosphorylation; silage; zearalenone.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Phylogenetic analysis based on core genome sequence alignments of S62-W and 39 Bacillus reference genomes.
Figure 2
Figure 2
HPLC chromatogram of (A) S62-W culture spiked with 50 µg/mL of ZEA (0 h), (B) S62-W culture spiked with 50 µg/mL of ZEA (24 h), and (C) S62-W culture without ZEA (24 h).
Figure 3
Figure 3
HPLC UV spectra of ZEA and compound X.
Figure 4
Figure 4
MS spectra of (A) standard ZEA (25 µg/mL) and (B) compound X (25 µg/mL); MS/MS spectrum of (C) compound X.
Figure 4
Figure 4
MS spectra of (A) standard ZEA (25 µg/mL) and (B) compound X (25 µg/mL); MS/MS spectrum of (C) compound X.
Figure 5
Figure 5
Structure of ZEA-14-phosphate.
Figure 6
Figure 6
Correlation (A) between ZEA decrease and growth of Bacillus sp. S62-W in CMB at 28 °C (n = 3), and transformation (B) of ZEA and production of ZEA-14-phosphate by Bacillus sp. S62-W in CMB at 28 °C (n = 3).
Figure 7
Figure 7
(A): ZEA transformation by Bacillus sp. S62-W in various media (28 °C, 3 days, n = 3). CMB: Corn meal broth W/O yeast extract; ASE: Alfalfa silage extract; CSE: Corn silage extract. (B): ZEA transformation by Bacillus sp. S62-W in CMB with pH value between 3 and 10 (28 °C, 5 days, n = 3). (C): ZEA transformation by Bacillus sp. S62-W in CMB incubated between 22 and 42 °C (2 days, n = 3). (D): ZEA transformation by Bacillus sp. S62-W in CMB under different incubation conditions (n = 3). Aerobic: Aerobic incubation at 28 °C for 2 days; Anaerobic: Anaerobic incubation at 28 °C for 2 days; AN-A: Anaerobic incubation at 28 °C for 6 days following by aerobic incubation at 28 °C for 2 days. (E): ZEA transformation by Bacillus sp. S62-W in CMB spiked with concentration varying between 25 and 200 µg/mL of ZEA (28 °C, 24 h and 96 h, n = 3).
Figure 7
Figure 7
(A): ZEA transformation by Bacillus sp. S62-W in various media (28 °C, 3 days, n = 3). CMB: Corn meal broth W/O yeast extract; ASE: Alfalfa silage extract; CSE: Corn silage extract. (B): ZEA transformation by Bacillus sp. S62-W in CMB with pH value between 3 and 10 (28 °C, 5 days, n = 3). (C): ZEA transformation by Bacillus sp. S62-W in CMB incubated between 22 and 42 °C (2 days, n = 3). (D): ZEA transformation by Bacillus sp. S62-W in CMB under different incubation conditions (n = 3). Aerobic: Aerobic incubation at 28 °C for 2 days; Anaerobic: Anaerobic incubation at 28 °C for 2 days; AN-A: Anaerobic incubation at 28 °C for 6 days following by aerobic incubation at 28 °C for 2 days. (E): ZEA transformation by Bacillus sp. S62-W in CMB spiked with concentration varying between 25 and 200 µg/mL of ZEA (28 °C, 24 h and 96 h, n = 3).
Figure 7
Figure 7
(A): ZEA transformation by Bacillus sp. S62-W in various media (28 °C, 3 days, n = 3). CMB: Corn meal broth W/O yeast extract; ASE: Alfalfa silage extract; CSE: Corn silage extract. (B): ZEA transformation by Bacillus sp. S62-W in CMB with pH value between 3 and 10 (28 °C, 5 days, n = 3). (C): ZEA transformation by Bacillus sp. S62-W in CMB incubated between 22 and 42 °C (2 days, n = 3). (D): ZEA transformation by Bacillus sp. S62-W in CMB under different incubation conditions (n = 3). Aerobic: Aerobic incubation at 28 °C for 2 days; Anaerobic: Anaerobic incubation at 28 °C for 2 days; AN-A: Anaerobic incubation at 28 °C for 6 days following by aerobic incubation at 28 °C for 2 days. (E): ZEA transformation by Bacillus sp. S62-W in CMB spiked with concentration varying between 25 and 200 µg/mL of ZEA (28 °C, 24 h and 96 h, n = 3).
Figure 8
Figure 8
ZEA phosphorylation activity of 23 Bacillus spp. and Bacillus sp. S62-W in CMB at 28 °C for 24 h incubation.
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References

    1. Zinedine A., Soriano J.M., Moltó J.C., Mañes J. Review on the toxicity, occurrence, metabolism, detoxification, regulations and intake of zearalenone: An oestrogenic mycotoxin. Food Chem. Toxicol. 2007;45:1–18. doi: 10.1016/j.fct.2006.07.030. - DOI - PubMed
    1. Tanaka T., Hasegawa A., Yamamoto S., Lee U.S., Sugiura Y., Ueno Y. Worldwide contamination of cereals by the fusarium mycotoxins nivalenol, deoxynivalenol, and zearalenone. 1. Survey of 19 countries. J. Agric. Food Chem. 1988;36:979–983. doi: 10.1021/jf00083a019. - DOI
    1. Tangni E.K., Pussemier L., Van Hove F. Mycotoxin contaminating maize and grass silages for dairy cattle feeding: Current state and challenges. J. Anim. Sci. Adv. 2013;3:492–511.
    1. Ogunade I.M., Martinez-Tuppia C., Queiroz O.C.M., Jiang Y., Drouin P., Wu F., Vyas D., Adesogan A.T. Silage review: Mycotoxins in silage: Occurrence, effects, prevention, and mitigation. J. Dairy Sci. 2018;101:4034–4059. doi: 10.3168/jds.2017-13788. - DOI - PubMed
    1. EFSA Scientific opinion on the risks for public health related to the presence of zearalenone in food. EFSA J. 2011;9:2197. doi: 10.2903/j.efsa.2011.2197. - DOI

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