isopropylstilbene发酵条件优化及其对灰葡萄孢的抑制作用

Fermentation condition optimization of isopropylstilbene and its inhibition effect against Botrytis cinerea

isopropylstilbene (3, 5-二羟基-4-异丙基二苯乙烯)是发光杆菌Photohabdus temperata产生的一种二苯乙烯类化合物,具有多种生物活性。为促进isopropylstilbene在农业抗生素领域应用价值的开发,采用固相萃取、柱层析等技术从发光杆菌P. temperata SN 35的发酵液中分离纯化得到1个化合物,通过核磁共振氢谱对其结构进行了确认,并采用高效液相色谱测定了其纯度;通过Plackett-Burman试验和响应面法对发光杆菌SN 35摇瓶发酵培养条件进行了优化,并初步研究了所分离化合物对灰葡萄孢Botrytis cinerea的离体抑菌活性及对灰霉病的防治效果。结果表明:所分离的化合物为isopropylstilbene,纯度为93%;对发光杆菌SN 35摇瓶发酵培养条件进行优化后,isopropylstilbene的产量是优化前的2.22倍。离体抑菌试验结果表明:isopropylstilbene对灰葡萄孢的菌丝生长具有较好的抑制作用,其EC_(50)值为(9.17±0.17)μg/mL,但低于对照药剂百菌清[EC_(50)值为(0.90±0.08)μg/mL];在10μg/mL下,isopropylstilbene对灰葡萄孢孢子萌发的抑制率为63.01%。活体盆栽试验结果表明,在200μg/mL下,isopropylstilbene对黄瓜灰霉病的防治效果达92.19%,与百菌清的防效(97.63%)相当,对番茄灰霉病的防治效果为72.12%,略低于百菌清(81.61%)。本研究通过对isopropylstilbene发酵条件的优化,为其工业化生产奠定了理论基础。该化合物对灰霉病具有很好的防治效果,可为天然杀菌剂的研究与开发提供参考。

Isopropylstilbene(3,5-dihydroxy-4-isopropyl stilbene)is a stilbene compound produced by Photohabdus temperata,which has various biological activities.In order to investigate the potential application of isopropylstilbene in the field of agricultural antibiotics,this compound was isolated from the fermentation broth of Photohabdus temperata SN35 by solid phase extraction and and purified by column chromatography.Its structure was confirmed by 1 H NMR and the purity was determined by high performance liquid chromatography.The shake flask fermentation conditions of the Photohabdus temperata SN35 were optimized by the Plackett-Burman test and the response surface method.The in vitro and in vivo antifungal activities of the compound against Botrytis cinerea were studied.The results showed that the isolated compound was isopropylstilbene with the purity of 93%.The yield of isopropylstilbene was 2.22 times higher than that before the optimization.The in vitro test results showed that the EC50 value of isopropylstilbene against the mycelial growth of B.cinerea was(9.17±0.17)μg/mL,which was lower than that of the control,chlorothalonil[EC50 value(0.90±0.08)μg/mL].And at 10μg/mL,the inhibition rate against spore germination was 63.01%.The results of pot experiments showed that the control effect of isopropylstilbene on cucumber gray mould was 92.19%at200μg/mL,which was almost the same with the control effect of chlorothalonil(97.63%).The control effect on tomato gray mold was 72.12%,which slightly lower than that of chlorothalonil(81.61%).This study laid a theoretical foundation for the industrial production by optimizing the fermentation conditions of isopropylstilbene.The compound has a good control effect against gray mold which will be useful for the research and development of natural fungicides.