Binks P R, French C E, Nicklin S, Bruce N C
Institute of Biotechnology, University of Cambridge, United Kingdom.
Appl Environ Microbiol. 1996 Apr;62(4):1214-9. doi: 10.1128/aem.62.4.1214-1219.1996.
A mixed microbial culture capable of metabolizing the explosive pentaerythritol tetranitrate (PETN) was obtained from soil enrichments under aerobic and nitrogen-limiting conditions. A strain of Enterobacter cloacae, designated PB2, was isolated from this culture and was found to use PETN as a sole source of nitrogen for growth. Growth yields suggested that 2 to 3 mol of nitrogen was utilized per mol of PETN. The metabolites pentaerythritol dinitrate, 3-hydroxy-2,2-bis-[(nitrooxy)methyl]propanal, and 2,2-bis-[(nitrooxy)methyl]-propanedial were identified by mass spectrometry and 1H-nuclear magnetic resonance. An NADPH-dependent PETN reductase was isolated from cell extracts and shown to liberate nitrite from PETN, producing pentaerythritol tri- and dinitrates which were identified by mass spectrometry. PETN reductase was purified to apparent homogeneity by ion-exchange and affinity chromatography. The purified enzyme was found to be a monomeric flavoprotein with a M(r) of approximately 40,000, binding flavin mononucleotide noncovalently.
在有氧和氮限制条件下,从土壤富集培养物中获得了一种能够代谢炸药季戊四醇四硝酸酯(PETN)的混合微生物培养物。从该培养物中分离出一株阴沟肠杆菌,命名为PB2,发现它能够利用PETN作为唯一的氮源进行生长。生长产量表明,每摩尔PETN可利用2至3摩尔氮。通过质谱和1H-核磁共振鉴定了代谢产物季戊四醇二硝酸酯、3-羟基-2,2-双-[(硝基氧基)甲基]丙醛和2,2-双-[(硝基氧基)甲基]丙二醛。从细胞提取物中分离出一种依赖NADPH的PETN还原酶,该酶可从PETN中释放亚硝酸盐,生成季戊四醇三硝酸酯和二硝酸酯,通过质谱对其进行了鉴定。通过离子交换和亲和色谱将PETN还原酶纯化至表观纯一。发现纯化后的酶是一种单体黄素蛋白,相对分子质量约为40000,非共价结合黄素单核苷酸。