Extraction and purification of ustiloxin A from rice false smut balls by a combination of macroporous resin and high-spe
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(2020) 2:29
RESEARCH
Food Production, Processing and Nutrition
Open Access
Extraction and purification of ustiloxin A from rice false smut balls by a combination of macroporous resin and high-speed countercurrent chromatography Gang Wang1,2, Dan He1, Fengchun Zhao3, Junqiang Hu1, Yin-Won Lee4, Jianrong Shi1,2* and Jianhong Xu1,2*
Abstract Rice false smut is an emerging plant disease worldwide. Ustiloxin A (UstA) is the major mycotoxin found in rice false smut balls, which are fungal colonies in rice florets. In this study, a new method consisting of macroporous resin column chromatography and high-speed countercurrent chromatography (HSCCC) was developed for UstA separation. UstA was extracted by a 3.81% HCOOH solution and adsorbed by XAD-4 resin. UstA was then eluted by a 40% methanol solution supplemented with 0.1% trifluoroacetic acid (TFA). Further purification was achieved by HSCCC using a two-phase solvent system consisting of n-butanol/TFA/H2O (1/0.05/1, v/v/v). Under the optimized conditions, 225 mg of UstA was obtained with a purity of 97.39% in a single run, with a final recovery of 65.2%. An inhibitory effect on seed germination of wheat and maize caused by UstA was observed in a preliminary phytotoxicity assay. Keywords: Ustiloxin A , Rice false smut, Preparative purification, Macroporous resin, High-speed counter-current chromatography, Phytotoxicity assay
Introduction Rice false smut is a plant disease caused by the fungal pathogen Ustilaginoidea virens Takahashi, which is now rapidly emerging in rice cultivation worldwide (Qiu et al. 2019). The most noticeable symptom of rice false smut is the false smut balls that form on rice florets as orange or green bulbs (Fan et al. 2020). In addition to the yield loss caused by rice false smut, U. virens produces secondary metabolites called mycotoxins. There are two main types of mycotoxins in rice false smut balls, * Correspondence: [email protected]; [email protected] 1 Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology/Key Laboratory for Control Technology and Standard for Agro-product Safety and Quality, Ministry of Agriculture and Rural Affairs/Key Laboratory for Agro-product Safety Risk Evaluation (Nanjing), Ministry of Agriculture and Rural Affairs/ Collaborative Innovation Center for Modern Grain Circulation and Safety/ Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing, People’s Republic of China Full list of author information is available at the end of the article
namely, ustiloxins (Koiso et al. 1994) and ustilaginoidins (Meng et al. 2015). Ustiloxin A (Fig. 1), the main derivative of ustiloxins, has shown phytotoxicity against seed germination (Koiso et al. 1992) as well as cytotoxic activity on bovine brain tubulin in vitro (Ludueńa et al. 1994) and early life zebrafish in vivo (Hu et al. 2019). UstA is also capable of causing “Lupinosis”-like lesions in mice (Nakamura et al. 1994). Hu et al. (2020) reported that UstA is pr
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