Studies on Biosysnthesis of Insecticidal Indole Alkaloid Okaramines

单纯青霉（Penicillium simplicissimum） ATCC 90288生长在山核桃上，产生具有独特结构的杀虫吲哚类生物碱山核桃胺。为了阐明克拉明的生物遗传途径，我们开展了多项研究，得到了如下一些结果。1)为了喂入标记的化合物，测试了几种液体培养培养基。在含有甲醇提取液的Czapek培养基中，胚芽生长良好，胚芽胺产量较高。2)在培养液中加入标记色氨酸，分离卡拉明B。质谱分析表明，其吲哚部分来源于色氨酸。3)大菜籽油和大豆中存在诱导单孢霉ATCC 90288产卡拉胺的因素。建立了一种评价卡拉明产量的生物测定系统，并用该系统对这些因子进行了部分纯化。4)从ATCC 90288中分离得到一个卡拉明同源物，并对其结构进行了分析。5)对ATCC 90288合成的萜类化合物进行了研究，分离出一种新的齐墩烷型三萜。6)新土壤分离物acleaspergillus acleatus KF-428与新同源物okamines H和I一起产生okamines A和B。

Penicillium simplicissimum ATCC 90288, when grown on okara, produces insecticidal indole alkaloid okaramines, which has a unique structure. To clarify the biogenetic pathway of okaramines, several subjects were carried out, and some results are obtained as follows.1) To feed labelled compounds, several media for liquid culture were tested. Czapek medium exsibited good growth and good production of okaramines in the presence of methanol extract of okara.2) Labelled tryptophan was added to the liquid culture, and okaramine B was isolated. Mass spectrum of the isolated okaramine B strongly indicated that indole parts of okaramines are derived from tryptophan.3) Okara and soyabean are strongly suggested to have some factors which induce okaramine production by P.simplicissimum ATCC 90288. A bioassay system for evaluating okaramine production was established, and these factors were partially purified with this system.4) A okaramine congener, okaramine G,was isolated from ATCC 90288, and the structure was elucidated.5) Investigation of terpene-related compounds produced by ATCC 90288 led to the isolation of a new oleanane triterpene.6) A new soil isolate Aspergillus aculeatus KF-428 produced okaramines A and B together with new congeners okaramines H and I.