STRUCTURAL IDENTIFICATION OF AUTOINDUCER OF PHOTOBACTERIUM-FISCHERI LUCIFERASE
STRUCTURAL IDENTIFICATION OF AUTOINDUCER OF PHOTOBACTERIUM-FISCHERI LUCIFERASE
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DOI：
10.1021/bi00512a013
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发表时间：
1981-01-01
期刊：
影响因子：
2.9
通讯作者：
OPPENHEIMER, NJ
中科院分区：
文献类型：
--
作者：
EBERHARD, A;BURLINGAME, AL;OPPENHEIMER, NJ
在某些发光细菌菌株中，细菌荧光素酶的合成需要由细菌合成并排泄到培养基中的自诱导剂的阈值浓度。采用乙酸乙酯萃取、溶剂蒸发、乙醇-水混合萃取、硅胶层析、正相高效液相层析等方法从无细胞培养基中分离出菲氏弧菌（P. fischeri, Vibrio fischeri）菌株mh -1分泌的自诱导剂。最终产品的纯度为99%。通过高分辨率1H核磁共振光谱、红外光谱和高分辨率质谱分析确定了该自诱导剂的结构为N-（3-氧己醇）-3-氨基二氢-2(3H)-呋喃酮[或N-(. β。-ketocaproyl)高丝氨酸内酯)。自诱导剂水解生成的同型丝氨酸符合这种结构。将高丝氨酸内酯偶联至3-氧己酸钠的乙二醇缩醛，以弱酸性去除保护基团，得到外消旋体形式的合成自诱导剂；该合成材料显示出相应的生物活性。
Synthesis of bacterial luciferase in some strains of luminous bacteria requires a threshold concentration of an autoinducer synthesized by the bacteria and excreted into the medium. Autoinducer excreted by P. fischeri (Vibrio fischeri) strain MJ-1 was isolated from the cell-free medium by extraction with ethyl acetate, evaporation of solvent, workup with ethanol-water mixtures and silica gel chromatography, followed by normal-phase and then reverse-phase high-performance liquid chromatography. The final product was > 99% pure. The structure of the autoinducer as determined by high-resolution 1H NMR spectroscopy, IR spectroscopy and high-resolution mass spectrometry was N-(3-oxohexanoyl)-3-aminodihydro-2(3H)-furanone [or N-(.beta.-ketocaproyl)homoserine lactone]. The formation of homoserine by hydrolysis of the autoinducer was consistent with this structure. Synthetic autoinducer, obtained as a racemate, was prepared by coupling homoserine lactone to the ethylene glycol ketal of sodium 3-oxohexanoate, followed by mildly acidic removal of the protecting group; this synthetic material showed the appropriate biological activity.