De Bellis Palmira, Tristezza Mariana, Haidukowski Miriam, Fanelli Francesca, Sisto Angelo, Mulè Giuseppina, Grieco Francesco
Institute of Sciences of Food Production, National Research Council, Unit of Bari, via Amendola 122/O, Bari 70126, Italy.
Institute of Sciences of Food Production, National Research Council, Unit of Lecce, via Provinciale Lecce-Monteroni, Lecce 73100, Italy.
Toxins (Basel). 2015 Nov 27;7(12):5079-93. doi: 10.3390/toxins7124864.
Ochratoxin A (OTA) is a mycotoxin with a main nephrotoxic activity contaminating several foodstuffs. In the present report, five soil samples collected from OTA-contaminated vineyards were screened to isolate microorganisms able to biodegrade OTA. When cultivated in OTA-supplemented medium, OTA was converted in OTα by 225 bacterial isolates. To reveal clonal relationships between isolates, molecular typing by using an automated rep-PCR system was carried out, thus showing the presence of 27 different strains (rep-PCR profiles). The 16S-rRNA gene sequence analysis of an isolate representative of each rep-PCR profiles indicated that they belonged to five bacterial genera, namely Pseudomonas, Leclercia, Pantoea, Enterobacter, and Acinetobacter. However, further evaluation of OTA-degrading activity by the 27 strains revealed that only Acinetobacter calcoaceticus strain 396.1 and Acinetobacter sp. strain neg1, consistently conserved the above property; their further characterization showed that they were able to convert 82% and 91% OTA into OTα in six days at 24 °C, respectively. The presence of OTα, as the unique OTA-degradation product was confirmed by LC-HRMS. This is the first report on OTA biodegradation by bacterial strains isolated from agricultural soils and carried out under aerobic conditions and moderate temperatures. These microorganisms might be used to detoxify OTA-contaminated feed and could be a new source of gene(s) for the development of a novel enzymatic detoxification system.
赭曲霉毒素A(OTA)是一种主要具有肾毒性的霉菌毒素,会污染多种食品。在本报告中,对从受OTA污染的葡萄园采集的五个土壤样本进行了筛选,以分离出能够生物降解OTA的微生物。在添加了OTA的培养基中培养时,225株细菌分离株将OTA转化为OTα。为了揭示分离株之间的克隆关系,使用自动rep-PCR系统进行了分子分型,结果显示存在27种不同的菌株(rep-PCR图谱)。对每个rep-PCR图谱的代表性分离株进行16S-rRNA基因序列分析表明,它们属于五个细菌属,即假单胞菌属、勒克氏菌属、泛菌属、肠杆菌属和不动杆菌属。然而,对这27株菌株的OTA降解活性进行进一步评估后发现,只有醋酸钙不动杆菌菌株396.1和不动杆菌属菌株neg1始终保持上述特性;对它们的进一步表征表明,在24℃下,它们能够在六天内分别将82%和91%的OTA转化为OTα。通过液相色谱-高分辨质谱法(LC-HRMS)确认了OTα作为唯一的OTA降解产物的存在。这是关于从农业土壤中分离的细菌菌株在有氧条件和适度温度下对OTA进行生物降解的首次报告。这些微生物可用于对受OTA污染的饲料进行解毒,并且可能是开发新型酶解毒系统的新基因来源。