Anderson R C, Rasmussen M A, Allison M J
Physiopathology Research Unit, U.S. Department of Agriculture, Ames, Iowa.
Appl Environ Microbiol. 1993 Sep;59(9):3056-61. doi: 10.1128/aem.59.9.3056-3061.1993.
The nitro toxins 3-nitro-1-propionic acid (NPA) and 3-nitro-1-propanol (NPOH), which are found in many leguminous plants, are known to be detoxified by ruminal microorganisms. The rates of the detoxification reactions are critical to acquisition of tolerance to the plants by ruminant animals, but further information is needed about factors which influence reaction rates and about the nature of the detoxification reactions. We found that rates of disappearance of NPA and NPOH varied somewhat between samples of ruminal fluid but were usually about 0.4 and 0.1 mumol/ml of ruminal fluid per h, respectively, and that rates with threefold-concentrated cells from rumen fluid were correspondingly higher. We present evidence that ruminal microbes from both cattle and sheep reduce these nitro groups in situ, so that NPA is converted to bet-alanine and NPOH is converted to 3-amino-1-propanol. These products were identified by thin-layer chromatography and, as their dabsyl derivatives, separated by high-performance liquid chromatography. The product beta-alanine was itself metabolized by these mixed suspensions of rumen microbes, so its recovery was always less than what would be estimated from NPA loss, but as much as 87% of the NPOH lost from incubation mixtures was recovered as 3-amino-1-propanol. Addition of sulfide and ferrous ions to suspensions of ruminal microbes increased the rate of NPOH reduction about threefold, but rates of NPA reduction were not similarly increased. When incubations were under hydrogen gas instead of carbon dioxide, the addition of sulfide and ferrous ions led to even greater (five- to eightfold) increases in the rates of NPOH metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)
在许多豆科植物中发现的硝基毒素3-硝基-1-丙酸(NPA)和3-硝基-1-丙醇(NPOH),已知可被瘤胃微生物解毒。解毒反应的速率对于反刍动物获得对这些植物的耐受性至关重要,但关于影响反应速率的因素以及解毒反应的性质,还需要更多信息。我们发现,瘤胃液样本中NPA和NPOH的消失速率略有不同,但通常分别约为每小时0.4和0.1 μmol/毫升瘤胃液,并且来自瘤胃液的三倍浓缩细胞的反应速率相应更高。我们提供的证据表明,牛和羊的瘤胃微生物都能在原位还原这些硝基,从而使NPA转化为β-丙氨酸,NPOH转化为3-氨基-1-丙醇。这些产物通过薄层色谱法进行鉴定,并作为它们的丹磺酰衍生物,通过高效液相色谱法进行分离。产物β-丙氨酸本身会被这些瘤胃微生物的混合悬液代谢,因此其回收率总是低于根据NPA损失估计的值,但从孵育混合物中损失的NPOH中,多达87%以3-氨基-1-丙醇的形式被回收。向瘤胃微生物悬液中添加硫化物和亚铁离子可使NPOH的还原速率提高约三倍,但NPA的还原速率没有类似增加。当在氢气而非二氧化碳条件下进行孵育时,添加硫化物和亚铁离子会使NPOH的代谢速率有更大幅度(五至八倍)的增加。(摘要截短于250字)