Isolation and structure elucidation of aerial mycelium-inhibitory substance in actinomycetes and differentiation mechanism of aerial mycelia

放线菌气生菌丝抑制物质的分离、结构解析及气生菌丝分化机制

• 批准号: 06660131
• 负责人: NATSUME Masahiro
• 金额: $ 1.34万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660131/
• 关键词: Streptomyces; Actinomycetes; morphological differentiation; Aerial mycelium-inhibitory substance

1.Aerial mycelium-inhibitory substance in Streptomyces alboniger Culture filtrate of S.alboniger in Bennet's medium was extracted with EtOAc at pH3 to give EtOAc soluble acidic fraction. The extract was purified by silica gel column and pTLC,and then fractionated by ODS-HPLC with the guidance of UV absorption at 215 nm. The peak at Rt 7.8 min showed aerial mycelium-inhibitory activity, however its activity became incomplete after ODS-HPLC.The active substance was identified as phenylacetic acid by spectral analysis. Addition of the peak fraction at Rt 4.7 min to phenylacetic acid recovered complete aerial mycelium inhibition. The synergistic substance at Rt 4.7 min was identified as 4-hydroxybenzaldehyde. Since phenylacetic acid is a metabolite of phenylalanine, the aerial mycelium-inhibitory activity of other related metabolites were examined. As a result, phenylethylamine and phenylacetaldehyde showed stronger activity than phenylalanine by itself. The structure-activity relationships of 4-hydroxybenzaldehyde was also examined.2.Improvement of carbazomycinal production Carbazomycinal is an aerial mycelium-inhibitory substance isolated from Streptoverticillium sp.by our group. For the mass production of carbazomycinal, we improved cultured conditions. Use of the production medium of carbazomycin antibiotics, which have carbazole nucleus similarly to carbazomycinal, and addition of antifoaming agentresulted in 56-fold yield of carbazomycinal.3.Screening of the strain that produce aerial mycelium-regulatory substance The strain that produce aerial mycelium-inhibitory or-inducing substance was surveyed in the culture collection of our laboratory. We have not found aerial mycelium inhibitor producing strain yet, but could found 3 strains that produce aerial mycelium-inducing substance.

1.用EtOAc在pH3条件下提取白链霉菌在Bennet培养基中的培养滤液，得到EtOAc可溶酸性组分。提取液经硅胶柱和薄层色谱纯化，在215 nm紫外吸收引导下，采用ODS-HPLC进行分离。在rt7.8 min时出现抑制气相菌丝的活性，但经ODS-HPLC后活性不完全。通过光谱分析鉴定其活性物质为苯乙酸。在4.7 min时加入苯基乙酸，可恢复对空气中菌丝的完全抑制。Rt 4.7 min时的增效物质为4-羟基苯甲醛。由于苯乙酸是苯丙氨酸的代谢物，因此对其他相关代谢物的抑气活性进行了研究。结果表明，苯乙胺和苯乙醛表现出比苯丙氨酸更强的活性。研究了4-羟基苯甲醛的构效关系。卡帕霉素是本课目从链霉菌中分离得到的一种气相菌丝抑制物质。为了卡帕霉素的大规模生产，我们改进了培养条件。2 .利用卡巴霉素类抗生素的生产培养基，卡巴霉素类抗生素具有与卡巴霉素相似的咔唑核，并加入消泡剂，使卡巴霉素的产率达到56倍。对本实验室收集的培养物中产生气生菌丝抑制或诱导物质的菌株进行了调查。我们还没有发现产生空中菌丝抑制剂的菌株，但发现了3株产生空中菌丝诱导物质的菌株。