O'Kane D J, Karle V A, Lee J
Biochemistry. 1985 Mar 12;24(6):1461-7. doi: 10.1021/bi00327a026.
Bright strains of the marine bioluminescent bacterium Photobacterium leiognathi produce a "lumazine protein" in amounts comparable to that previously found in Photobacterium phosphoreum. New protocols are developed for the purification to homogeneity of the proteins from both species in yields up to 60%. In dimmer strains the amounts of lumazine protein in extracts are less, and also there is an accompanying shift of the bioluminescence spectral maximum to longer wavelength, 492 nm. Both types of lumazine proteins have identical fluorescence spectra, with maxima at 475 nm, so it is suggested that, whereas lumazine protein is the major emitter in bright strains, there is a second emitter also present with a fluorescence maximum at longer wavelength. The two species of lumazine protein have the same 276 nm/visible absorbance ratio, 2.2, but differ in visible maxima: P. phosphoreum, 417 nm; P. leiognathi, 420 nm. For the latter the bound lumazine has epsilon 420 = 10 100 M-1 cm-1, practically the same as in free solution. The two lumazine proteins also differ quantitatively in their effect on the in vitro bioluminescence reaction, i.e., at blue shifting the bioluminescence spectrum or altering the kinetics. The P. phosphoreum lumazine protein is more effective with its homologous luciferase or with P. leiognathi luciferase than is the lumazine protein from P. leiognathi. These differences may have an electrostatic origin.
明亮型的海洋发光细菌利氏发光杆菌（Photobacterium leiognathi）产生的“鲁玛嗪蛋白”数量与先前在磷光发光杆菌（Photobacterium phosphoreum）中发现的数量相当。已开发出新的方案，用于从这两个物种中纯化蛋白质至同质，产率高达60%。在较暗的菌株中，提取物中鲁玛嗪蛋白的量较少，并且生物发光光谱最大值也伴随有向更长波长（492nm）的偏移。两种类型的鲁玛嗪蛋白具有相同的荧光光谱，最大值在475nm，因此有人提出，虽然鲁玛嗪蛋白是明亮菌株中的主要发光体，但也存在另一种发射体，其荧光最大值在更长波长处。这两种鲁玛嗪蛋白具有相同的276nm/可见光吸光度比，即2.2，但可见光最大值不同：磷光发光杆菌为417nm；利氏发光杆菌为420nm。对于后者，结合的鲁玛嗪的ε420 = 10 100 M-1 cm-1，实际上与在自由溶液中的相同。这两种鲁玛嗪蛋白在对体外生物发光反应的影响上也存在定量差异，即在使生物发光光谱蓝移或改变动力学方面。磷光发光杆菌的鲁玛嗪蛋白与其同源荧光素酶或利氏发光杆菌荧光素酶相比，比利氏发光杆菌的鲁玛嗪蛋白更有效。这些差异可能源于静电作用。
