Lampteromyces Bioluminescence---production of the light emitter from riboflavin

本项目将对月夜菇（TSUKIYOTAKE）生物发光的分子机制进行研究。根据生物发光光谱在524 nm处显示其最大值和提取物显示相同的荧光光谱，该发光的发光体的发色团被指定为异alloxazine。培养发光菌丝体，使其表现出与蘑菇鳃相同的光。这种发光物质被命名为灯黄素。它是从酸性溶液中分离出来的，在酸性溶液中，新鲜的鳃浸泡在氧气气氛下发光过夜。溶液通过结合疏水物质的柱。甲醇洗脱液产生一个单一的绿色荧光化合物灯黄素，在FAB-MS光谱中显示为m/ z509。灯黄素易被酸水解成核黄素，m/x 377；质量差表明存在戊二烯。采用串联质谱法，以m/ z259为亲本离子，对该糖进行了鉴定。通过4个真实的MS/MS谱图的比较，鉴定该糖为核糖。核磁共振谱进一步证实了这一点。NOE实验揭示了核黄素5'位置α链上的核糖的连接。水解产物的CD谱显示d -核糖；因此，灯黄素的绝对结构是5'- - d -核糖呋喃基核黄素。下一步将进行立体选择性合成灯黄素的研究，为研究蘑菇生物发光的分子机制提供依据。

The molecular mechanism of the bioluminescence in moon-night mushroom (TSUKIYOTAKE) is to be elucidated in this project. The chromophore of the light emitter in this luminescence was assigned to be isoalloxazine from the facts that the bioluminescence spectrum showed its maximum at 524 nm and that the extracts showed an identical fluorescence spectrum. Luminous myceria was cultivated to exhibit the same light as the gills from the mushroom. The light emitter was named as lampteroflavin. It was isolated from an acidic solution in which fresh gills were immerged and luminesced for overnight under oxygen atmosphere. The solution was passed through columns which combined the hydrophobic substances. Methanol eluate afforded a single green fluorescent compound, lampteroflavin, which showed m/z 509 in FAB-MS spectrum. Lampteroflavin was readily hydrolyzed by acid into riboflavin, m/x 377; the mass difference 133 suggested the presence of pentse. This sugar was identified by tandem MS/MS of hexaacetate of Lf at m/z 259 as parent ion. From comparison of four authentic MS/MS spectra the sugar was identified to be ribose. This was further supported from nmr spectra. NOE experiments disclosed the connection of the ribose in alpha linkage at the 5' position of riboflavin. CD spectrum of the hydrolysate concluded the D-ribose; and thus the absolute structure of lampteroflavin is 5'- -D-ribofuranosyl-riboflavin. Further studies will be followed by stereoselective synthesis of lampteroflavin to supply for the investigation of the molecular mechanism of the mushroom bioluminescence.