Preparation/production and Utilization Researches of Terpenoidal Plant-growth Stimulants and Other Bioactive Substances with High Bioactivity.

萜类植物生长刺激剂及其他高生物活性物质的制备/生产及利用研究。

• 批准号: 12556015
• 负责人: SASSA Takeshi
• 金额: $ 8.38万
• 依托单位: Yamagata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12556015/
• 关键词: fluorofusicoccin; semi-synthesis; Phomopsis amygdali; fusicoccin; cotylenin; seed-germination promotion; cell differentiation; antitumor; 発酵生産; 白血病細胞; 分化誘導; 植物成長調節; アブシジン酸; 3-エピフシコクシン; ガン細胞; ABA; Phomopsis amygdali; フロロフシコクシン; コチレノール; フ

1.Preparation of Highly Bioactive Fusicoccin Derivatives by Reverse-order Semi-synthesisWe have succeeded in obtaining 19/20-fluorofusicoccins quite efficiently by synthesizing 19/20-fluoro derivatives of fusicocca-2,10(14)-diene, a genuine biosynthetic intermediate of fusicoccins, and then feeding them into the fusicoccin-producing fungus for metabolic transformations. We named this novel process a "Reverse-order Semi-synthesis", as the approach deviates reversely from typical semi-synthesis insofar as purely chemical steps are followed by biochemical transformations.2.Finding of New Fusicoccin-producing Fungi in Japan and Gram-scale Production of FusicossinsWe found peach Fusicoccum canker fungi (Phomopsis amygdali Niigata) as the first fusicoccin-producing fungi in Japan, and selected its Niigata 74 strain as one of the highest fusicoccin-productive strains. This strain in a commercial sugar-soybean meal medium produced fusicoccin A and 3'-deacetylfusicoccin A (each 0.4 g/L) for its one-week cultivation or only 3'-deacetylfusicoccin A (0.9g/L) for its 2-week cultivation.3.Isolation of New Fusicoccins and Their Seed Germination-stimulating ActivityFrom the strain Niigata 74 new fusicoccins P, Q, R, and S, and 3-epifusicoccins H and Q were isolated as polar fusicoccins, and their structures were elucidated by chemical and spectroscopic evidence. Fusicoccins P and R being 16-O-methylfusicoccins showed the high activity in the seed germination-stimulating test in the presence of abscisic acid.4.Mammal Cell Differentiation-inducing Activity of Cotylenin A and Its Tumor Cell-Growth Suppressive ActivityCotylenin A induced the differentiation of human acute myeloid leukaemia cells in primaiy culture, and showed neurite genaration (outgrowth) promoting activity toward PC12 nerve cells at low concentrations. Cotylenin A and interferon-α cooperatively induced apoptosis and have an antitumor effect on human non-small cell lung carcinoma cells in nude mice.

1.通过逆序半合成制备高生物活性的Fusicoccin衍生物我们已经成功地通过合成Fusicocca-2，10（14）-二烯的19/20-氟衍生物（一种真正的Fusicoccin的生物合成中间体）非常有效地获得了19/20-氟Fusicoccin，然后将它们喂入产生Fusicoccin的真菌进行代谢转化。我们将这一新的过程命名为“逆序半合成”，因为这种方法与典型的半合成方法有很大的不同，因为在纯化学步骤之后是生化转化。2.在日本发现新的产梭孢菌素的真菌和革兰氏规模生产梭孢菌素我们发现了桃梭孢菌素溃疡菌（Phomopsis amygdali新泻）作为日本第一株产梭菌素的真菌，筛选出其产梭菌素最高的菌株之一新泻74。该菌株在商业糖-大豆粉培养基中产生梭孢菌素A和3 ′-脱乙酰梭孢菌素A（各0.4g/L）进行一周培养或仅3 '-脱乙酰梭孢菌素A（0.9g/L）培养2周。3.新的丝孢菌素的分离及其种子萌发刺激活性从菌株新泻中分离得到74个新的丝孢菌素P、Q、R和S，3-epifusicoccins H和Q为极性fusicoccins，经化学和光谱鉴定其结构。梭孢菌素P和R是16-O-甲基梭孢菌素，在脱落酸存在下在种子萌发刺激试验中显示出高活性。4. Cotylenin A的哺乳动物细胞分化诱导活性及其肿瘤细胞生长抑制活性Cotylenin A在原代培养中诱导人急性髓性白血病细胞分化，并且在低浓度下显示出对PC 12神经细胞的神经突生成（生长）促进活性。Cotylenin A和IFN-α协同诱导裸鼠人非小细胞肺癌细胞凋亡及抗肿瘤作用

项目成果

T.Sassa: "Fusicoccins P and Q, and 3-epifusicoccins H and Q, new polar fusicoccins from isolate Niigata 2-A of a peach Fusicoccum canker fungus."Biosci.Biotechnol.Biochem.. 66(11). 2356-2361 (2002)

T.Sassa：“Fusicoccins P 和 Q，以及 3-epifusicoccins H 和 Q，来自桃 Fusicocum 溃疡病真菌 Niigata 2-A 分离株的新型极性 Fusicoccins。”Biosci.Biotechnol.Biochem.. 66(11)。

N.Tajima: "Chemical identification of fusicoccins from a Japanese isolate Niigata 2 of peach Fusicoccum canker fungus (Phomopsis amygdoli) and production of 3'-deacetylfusicoccinA by the fungus."J.Pesticides Sci.. 27(1). 64-67 (2002)

N.Tajima：“对桃褐霉病菌（Phomopsis amygdoli）的日本分离株 Niigata 2 中的梭菌素进行化学鉴定，并通过该真菌生产 3-脱乙酰梭菌素 A。”J.Pesticides Sci. 27(1)。

K.Yamada: "Differentiation of human acute myeloid leukaemia cells in response to cotylenin A, a plant growth regulator."J.Haematology. 114. 814-821 (2001)

K.Yamada：“人急性髓系白血病细胞对植物生长调节剂子叶素 A 的反应的分化。”J.Haematology。

Y.Honma: "Cotylenin A, a differentiation-inducing agent, and IFN-α cooperatively induce apoptosis and have an antitumor effect on human non-small cell lung carcinoma cells in nude mice."Cancer Res.. 63. 3659-3666 (2003)

Y.Honma：“分化诱导剂 Cotylenin A 和 IFN-α 协同诱导细胞凋亡，并对裸鼠体内的人非小细胞肺癌细胞具有抗肿瘤作用。”Cancer Res.. 63. 3659-3666 ( 2003）

加藤 修雄: "特異な生理活性菌頚ジテルペン配糖体-フシコクシン、エリナシン-における初期生合成経路の解明と応用"日本農芸化学会誌. 76・12. 1202-1205 (2002)

加藤伸夫：“独特生理活性真菌二萜糖苷 - 梭菌素和埃里那星的初始生物合成途径的阐明和应用”，日本农业化学学会杂志 76・12（2002 年）。