一株脱氧雪腐镰刀菌烯醇生物转化菌的分离,鉴定和转化效果的初步研究
首发时间:2007-02-15
摘要:以脱氧雪腐镰刀菌烯醇(DON)为唯一碳源,通过富集培养从土壤中分离到一株可以生物转化DON的真菌,编号为A-DON。用真菌引物Bt2a和Bt2b扩增了该菌的β-tubulin 基因序列,通过β-tubulin 基因序列分析和形态学观察最终鉴定该菌株为塔宾曲霉(Aspergillus tubingensis)。该菌的β-tubulin 基因序列在GenBank的登录号为DQ902579,其与塔宾曲霉AY820009的相似性为99%。在pH6,30℃,14d内其对DON的平均转化率为94.4%。通过对DON的转化产物的HPLC-MS-MS分析,结果表明转化产物的分子量为314.4D,与DON的分子量(296.3D)相比增加了18.1D,其机理可能是DON的主要毒性基团12,13-环氧环打开变成了两个羟基,其分子量就成了314.4D。该菌在无机盐中对DON的耐受浓度高为40mg/L,可望用于霉败饲料的脱毒,为DON的生物转化提供了一种新的途径。
关键词: 脱氧雪腐镰刀菌烯醇 生物转化 分离 鉴定 β-tubulin基因
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Isolation and Identification of a Strain of Aspergillus tubingensis With Deoxynivalenol Biotransformation Capability
Abstract:A colony was isolated from soil by inorganic salt culture media added (deoxynivalenol)DON and numbered A-DON. Bt2a-Bt2b were used as primers to amplify the β-tubulin gene of A-DON. Sequence analysis and morphology observation indicated that A-DON belonged to Aspergillus tubingensis. Sequence analysis of Bt2a-Bt2b was submitted to GenBank. The access number in GenBank was DQ902579. Its homology to Aspergillus tubingensis AY820009 was 99%. The mean DON-biotransformation rate was 94.4% after two weeks cultivation. The molecular weight of biotransformation product analyzed by HPLC-MS/MS was 314.4 D which was different from molecular weight of DON (296.3 D). The molecular weight of biotransformation product was 18.1 D larger than that of DON, which indicated that 12,13-epoxy ring in DON could be changed into 12,13-hydroxide radical. It indicated that the A-DON had the ability to transform DON into another product. A-DON could tolerate 40mg L-1 DON in inorganic salt media. It could to be used for detoxification of mycotoxin in mouldy food and feedstock. The research exhibited a new method for DON.biotransformation.
Keywords: Deoxynivalenol Isolation Identification β-tubulin gene Biotransformation
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一株脱氧雪腐镰刀菌烯醇生物转化菌的分离,鉴定和转化效果的初步研究
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