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冠菌素(coronatine,COR)是一种环境友好的新型植物生长调节剂,能有效调控植物生长,增强植物抗逆性。目前冠菌素主要通过微生物发酵的方式获得,而发酵效率低是限制冠菌素应用的最大问题。为提高冠菌素产量,本研究建立了丁香假单胞菌(Pseudomonas syringae)Z2-6的半连续发酵方式,并对移取起始时间、移取间隔时间以及移取体积3个方面进行了优化。结果表明,第10天开始移取较第8、9天开始移取可以延长冠菌素产出周期,使冠菌素合成始终维持在较高的水平。间隔1 d移取较间隔2 d移取冠菌素生产效率更高。当移取量不大于30%时,不会对菌体量产生影响,并且当移取量为30%时冠菌素产量达到最高。与传统发酵方法相比,在发酵第10天起每隔1 d移出30%发酵液并补充等体积新鲜培养基,可使冠菌素产量由11.2 mg/(L·d)提高到22.1 mg/(L·d),生产效率提高了98%。本研究利用半连续发酵方式有效提高了冠菌素发酵生产效率,节约了生产成本,为冠菌素的工业化生产和农业推广应用提供有力支持。
Coronatine (COR) is a new environment-friendly plant growth regulator, which can effectively regulate plant growth and enhance plant stress resistance. At present, coronatine is mainly obtained by means of microbial fermentation, and the low fermentation efficiency is the biggest problem that limits the application of coronatine. In order to improve the yield of coronatine, a semi-continuous fermentation method of Z2-6 of Pseudomonas syringae was established in this study, and three aspects of the start-up time, removal time and removal volume were established optimization. The results showed that the removal of coronatine could be prolonged on the 10th day and started on the 8th and 9th day, and the synthesis of coronatine was always maintained at a higher level. It is more efficient to remove coronatine for 1 day at an interval of 2 days. When the amount of removal is not more than 30%, the amount of bacteria will not be affected, and the production of coronatine reaches the highest when the amount of removal is 30%. Compared with the traditional fermentation method, 30% of fermentation broth was removed every 1 day from the 10th day of fermentation and the equal volume of fresh medium was supplemented to increase the production of coronatine from 11.2 mg / (L · d) to 22.1 mg / (L · d), production efficiency increased by 98%. In this study, the semi-continuous fermentation method was used to effectively improve the productivity of coronatine fermentation, save the production cost and provide strong support for the industrial production of Coronatine and agricultural extension and application.