Völkl Andrea, Vogler Bernhard, Schollenberger Margit, Karlovsky Petr
Max-Planck-Institut für Entwicklungsbiologie, Spemannstr. 35, D-72076 Tübingen, Germany.
J Basic Microbiol. 2004;44(2):147-56. doi: 10.1002/jobm.200310353.
Deoxynivalenol (DON) is a trichothecene secondary metabolite produced by Fusarium species infecting cereal crops. As a mycotoxin, DON causes losses in livestock production and poses a health risk to humans consuming contaminated cereal products. DON also acts as a virulence factor, facilitating the colonization of host plants by Fusarium spp. Enzymatic detoxification of mycotoxins in feed additives and genetically modified crops is a promising approach for the reduction of mycotoxin contamination of feeds and food. A prerequisite for the development of biotechnological strategies for DON detoxification is the availability of genes encoding suitable enzymatic activities. With the goal of isolating microbial cultures that can be used as a source of such activities, we screened 1285 microbial cultures from farmland soil, cereal grains, insects and other sources for DON transformation under aerobic conditions. One mixed culture transformed DON into two chromatographically separable products. The main product of the transformation was purified and its structure was elucidated by mass spectroscopy, (1)H-NMR, (13)C-NMR and proton-proton and carbon-proton correlated NMR spectroscopy. The structure of this product was determined to be 3-keto-4-deoxynivalenol. The DON-transforming mixed culture survived and retained its transforming activity during a starvation period of six months at 20 degrees C. Transformation of DON was suppressed by low concentrations of glucose and high concentrations of tryptone and yeast extract. Cell-free supernatants obtained either by filtration through a 0.22 microm membrane filter or by centrifugation did not exert DON-transforming activity. Trichothecenes 15-acetyl-DON, 3-acetyl-DON and fusarenon-X were also transformed.
脱氧雪腐镰刀菌烯醇(DON)是由感染谷类作物的镰刀菌属产生的一种单端孢霉烯族次生代谢产物。作为一种霉菌毒素,DON会导致畜牧生产损失,并对食用受污染谷类产品的人类健康构成风险。DON还作为一种毒力因子,促进镰刀菌属在宿主植物上的定殖。饲料添加剂和转基因作物中霉菌毒素的酶解毒是减少饲料和食品中霉菌毒素污染的一种有前景的方法。开发DON解毒生物技术策略的一个先决条件是要有编码合适酶活性的基因。为了分离可作为此类活性来源的微生物培养物,我们在有氧条件下从农田土壤、谷物、昆虫和其他来源筛选了1285种微生物培养物以进行DON转化。一种混合培养物将DON转化为两种色谱可分离的产物。转化的主要产物被纯化,其结构通过质谱、(1)H-NMR、(13)C-NMR以及质子-质子和碳-质子相关NMR光谱进行了阐明。该产物的结构被确定为3-酮-4-脱氧雪腐镰刀菌烯醇。在20摄氏度的饥饿期6个月期间,DON转化混合培养物存活并保留了其转化活性。低浓度葡萄糖、高浓度胰蛋白胨和酵母提取物会抑制DON的转化。通过0.22微米膜过滤器过滤或离心获得的无细胞上清液不具有DON转化活性。单端孢霉烯族毒素15-乙酰-DON、3-乙酰-DON和镰刀菌烯酮-X也被转化。