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首页> 外文期刊>Biochemistry >Compound 800,a Natural Product Isolated from Genetically Engineered Pseudomonas:Proposed Structure,Reactivity,and Putative Relation to Heme
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Compound 800,a Natural Product Isolated from Genetically Engineered Pseudomonas:Proposed Structure,Reactivity,and Putative Relation to Heme

机译:化合物800,一种从基因工程假单胞菌中分离得到的天然产物:拟议的结构,反应性和与血红素的推定关系

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Genetically engineered strains of Escherichia coli and Pseudomonas aeruginosa were prepared harboring the gene cluster nirFDLGH from Pseudomonas stutzeri substrain ZoBell on a high copy plasmid.These genes have been previously implicated as being essential for the biosynthesis of heme d_1,the prosthetic group of dissimilatory nitrite reductases in anaerobic,denitryfying bacteria.Tetrapyrroles detectable at steady-state levels were identified from both organisms,and cell-free extracts from each were also used to transform uroporphyrinogen in vitro.E.coli does not naturally produce d_1,and the engineered strain failed to produce d]or any tetrapyrrole foreign to E.coli.Therefore,while nirFDLGH may be necessary for d_1 biosynthesis,it is not sufficient.In the denitrifier P.aeruginosa,the results were more positive.The presence of the plasmid led to increased levels of d_1.In addition,a previously unidentified tetrapyrrole was detected.This compound was characterized by visible absorption spectroscopy,infrared spectroscopy,X-ray photoelectron spectroscopy,mass spectrometry,and NMR,and a tentative structure was proposed for this compound.The tetrapyrrole has structural features similar to sirohydrochlorin(as precorrin-2 or sirotetrahydrochlorin,a known intermediate of d_1)and d_1 itself.The most unusual substituents are epoxide and sulfoxide moieties.When this tetrapyrrole was treated with strong mineral acid and heat,it was converted into natural d_1.
机译:用高拷贝质粒制备了带有斯图氏假单胞菌亚种ZoBell基因簇nirFDLGH的大肠杆菌和铜绿假单胞菌的基因工程菌株。从这两种生物中鉴定出在稳态水平下可检测到的四环戊四醇,并且还将每种生物的无细胞提取物用于体外转化尿卟啉原。大肠杆菌不能自然地产生d_1,工程菌无法因此,虽然d_1生物合成可能需要nirFDLGH,但这还不够。在反硝化假单胞菌P.aeruginosa中,结果更为阳性。质粒的存在导致水平提高此外,还检测到先前未鉴定的四吡咯。该化合物的特征在于可见吸收s提出了该化合物的光谱学,红外光谱,X射线光电子能谱,质谱和NMR以及暂定结构。四吡咯的结构特征与西罗盐酸二氢卟酚(precorrin-2或sirotetrahydrochlorin,已知的d_1中间体)相似。 d_1本身。最不常见的取代基是环氧基和亚砜部分。当四吡咯经强无机酸和热处理后,会转化为天然d_1。

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