玉米赤霉烯酮降解菌的筛选及其活性检测

doi:10.11843/j.issn.0366-6964.2017.04.020

摘要/Abstract

摘要：

玉米赤霉烯酮（Zearalenone，ZEN）是危害动物健康的重要真菌毒素之一，笔者旨在筛选出能够有效降解ZEN的微生物，并对其降解特性以及降解产物的致毒活性进行检测，以期筛选出高效降解ZEN的菌株。以ZEN为唯一碳源进行初筛，以菌株发酵液对ZEN的降解率为复筛指标，对降解率最高的菌株进行16S rRNA基因测序分析；并对筛选出的菌株对ZEN的降解特性进行研究，对其降解产物在MCF-7细胞上进行活性检测。初筛获得40株能生长良好的细菌，复筛筛选出降解率最高H6菌株，经16S rRNA基因测序分析，该菌株为解淀粉芽孢杆菌（Bacillus amyloliquefaciens）。在37℃、180 r·min^-1、72 h时，该菌株发酵液对1 μg·mL^-1ZEN的降解率为95.6%，其上清液和菌悬液的降解率分别为68.93%和28.70%，对20 μg·mL^-1 ZEN的降解率为85.8%；菌株上清液经蛋白酶K、蛋白酶K+SDS和热处理后，其降解活性明显降低；该菌株ZEN的降解产物在MCF-7细胞上的增殖率显著降低。结果表明，本研究筛选出能有效降解ZEN的菌株H6，经鉴定判断为解淀粉芽孢杆菌（B. amyloliquefaciens），该菌株分泌的胞外酶能够降解ZEN，使其类雌激素毒性显著降低。

Abstract:

Zearalenone(ZEN) is one of the main mycotoxin endangering animal health. The aim of this research was to screen bacteria with the ability to degrade ZEN efficiently, and detect their degradation characteristics and toxicity. ZEN was used as the sole carbon source for the preliminary screening. And then the strains fermentation broth degradation rate was as the index for the second selection. The 16S rRNA sequence analysis was used to identify the strain with the highest ZEN degradation rate. The ZEN degradation characteristics of the obtained strain was studied, and the activity of the ZEN degradation product was tested in MCF-7 cells. Forty strains were screened out in the preliminary experiment, and strain H6 with the highest ZEN degradation rate was selected after the secondary screening. Strain H6 was identified as the Bacillus amyloliquefaciens by 16S rRNA sequence analysis. At 37℃, 180 r·min^-1, 72 h, the degradation rates of strain H6 fermentation liquor to 1 and 20 μg·mL^-1 ZEN were 95.6% and 85.8%, respectively. Among that, the degradation rates of the culture supernatant and cells to 1 μg·mL^-1 ZEN were 68.93% and 28.70%, respectively. The ZEN degradation activity of strain H6 culture supernatant was significantly decreased after treated with proteinase K, proteinase K combined with SDS and heating, respectively. Compared with the control group, the proliferation rate of MCF-7 cell was significantly reduced when treated with ZEN degradation product. In conclusion, strain H6 with the highest ZEN degradation rate was identified as Bacillus amyloliquefaciens by 16S rRNA sequence analysis. The extracellular enzymes secreted by strain H6 could degrade ZEN, and significantly reduce the estrogen activity.

中图分类号:

S852.6

魏单平, 刘玉洁, 孙向丽, 闫峰宾, 蒋瑞瑞, 康相涛, 王彦彬. 玉米赤霉烯酮降解菌的筛选及其活性检测[J]. 畜牧兽医学报, 2017, 48(4): 761-768.

WEI Dan-ping, LIU Yu-jie, SUN Xiang-li, YAN Feng-bin, JIANG Rui-rui, KANG Xiang-tao, WANG Yan-bin. Screening and Activity Detection of a Bacterial Strain Degrading Zearalenone[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(4): 761-768.

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