Shibata H, Odahara Y, Sasaki F, Hirota O, Nishida M, Tani I
Faculty of Pharmaceutical Sciences, University of Tokushima, Japan.
Microbiol Immunol. 1994;38(7):519-25. doi: 10.1111/j.1348-0421.1994.tb01817.x.
An extract from intact spores of Bacillus cereus T having a germination-inducing activity was studied. Two distinct germinative principles were found through dialysis of the extract. One was diffusible through the dialysis membrane and the other was non-diffusible. The activity of the former fraction was inhibited by the addition of 1 mM glycoletherdiamine-N,N,N',N'-tetraacetic acid (GEDTA), whereas the latter fraction was inactive unless GEDTA was added to the assay system. The diffusible principle maintained the major portion of the activity found in the crude spore extract. By means of high-performance liquid chromatography (HPLC) using a gel permeation chromatography column, 9 fractions were obtained from the deproteinized diffusible fraction. Of those fractions, two fractions (No. 1 and No. 8) were responsible for the germination-inducing activity, but no reconstituted activity was observed unless both fractions No. 1 and No. 8 were added to the assay system. Amino acid analysis of fraction No. 1 revealed that the fraction was rich in free amino acids, especially in alanine. On the other hand, by the use of reverse-phase HPLC and fast atom bombardment mass spectrometry, it was concluded that the effective substance in fraction No. 8 was inosine. Based on these findings, it was suggested that the active substances in fraction No. 1 might be a free amino acid such as L-alanine and/or Ca2+ and a Ca(2+)-binding substance.
对具有萌发诱导活性的蜡样芽孢杆菌T完整孢子提取物进行了研究。通过对提取物进行透析，发现了两种不同的萌发诱导成分。一种可透过透析膜，另一种则不能透过。向提取物中添加1 mM乙二醇醚二胺-N,N,N',N'-四乙酸（GEDTA）可抑制前一种成分的活性，而后一种成分在测定体系中不添加GEDTA时则无活性。可扩散成分保留了粗孢子提取物中大部分的活性。使用凝胶渗透色谱柱通过高效液相色谱（HPLC）从脱蛋白的可扩散成分中获得了9个组分。在这些组分中，有两个组分（第1号和第8号）具有萌发诱导活性，但除非将第1号和第8号组分都添加到测定体系中，否则未观察到重新组合后的活性。对第1号组分的氨基酸分析表明，该组分富含游离氨基酸，尤其是丙氨酸。另一方面，通过反相HPLC和快原子轰击质谱分析得出，第8号组分中的有效物质是肌苷。基于这些发现，有人提出第1号组分中的活性物质可能是诸如L-丙氨酸之类的游离氨基酸和/或Ca2+以及一种Ca(2+)结合物质。
