植物内生菌发酵培养基的初探

A study on fermentation media of plant endophytes

从高活性菌株到获得高产量活性产物之间,发酵是一个重要环节。本研究设计4种内生菌发酵培养基,对89株分离自傣药植物且具有高抗菌活性的内生菌及5株潜在放线菌新种进行摇瓶发酵,发酵液的提取物用3种展层系统作薄层色谱(TLC)展层,用UV(254nm、366nm)和茴香醛试剂进行检测。实验结果表明,用1号和4号培养基发酵产生的次生代谢产物的种类最丰富,2号和3号培养基产生的较少;用II号展层系统(氯仿∶甲醇=4∶1)和III号展层系统(正丁醇∶乙酸∶水=4∶1∶5)作TLC层析,检测到的产物最多;3种检测方法以茴香醛检测到的代谢产物最丰富。对每一株菌而言,产生最多产物的发酵培养基各不相同,为了获得尽可能多的次生代谢产物,不同菌株选用的发酵培养基不一样。探究适合的发酵培养条件,为活性产物的分离、制备奠定了基础。

Fermentation of 89 endophytes and 5 novel species candidates of actinomycetes were carried out by four different fermentation media. Three developing systems were used for chromatography of TLC. UV 254nm, 366nm and anisaldehyde as a chromogenic agent were used for detecting compounds of these strains. Results indicate that the metabolites of these strains were the most abundant when these strains were fermented in No. 1 and No. 4 media, and only few in No. 2 and No. 3. Metabolites of each strain were different in different media. More metabolites could be detected when the TLC was carried out in system Ⅱ （chloroform ： methanol=9 ： 1） and system Ⅲ （butanol ： acetic acid ： water=4 ： 1 ： 5） with anisaldehyde test solution. These result established a base for designing of fermentation medium, isolation and purification of effective compounds for discovery of new leader compounds from endophytes.