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

本项目旨在阐明月夜菇生物发光的分子机制。根据生物发光光谱在524 nm处显示其最大值以及提取物显示相同的荧光光谱的事实，将该发光中的发光体的发色团指定为异咯嗪。发光菌丝被培养成与蘑菇的鳃一样的光。发光体被命名为lampterflavin。将新鲜鳃浸入酸性溶液中，在氧气气氛下发光过夜。使溶液通过合并疏水物质的柱。甲醇洗脱得到单一的绿色荧光化合物灯黄素，其在FAB-MS谱中显示m/z 509。Lampteroflavin很容易被酸水解成核黄素，m/x 377;质量差133表明存在pentse。该糖通过Lf的六乙酸酯的串联MS/MS在m/z 259处作为母离子来鉴定。通过比较四个真实的MS/MS谱，确定糖是核糖。核磁共振谱进一步支持了这一点。NOE实验公开了核糖在核黄素的5'位以α键连接。水解产物的CD光谱推断为D-核糖;因此灯黄素的绝对结构为5 '-D-呋喃核糖基-核黄素。本研究将进一步立体选择性合成灯黄素，为蘑菇发光的分子机理研究提供理论依据。

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.