高产武夷菌素wysR基因过表达菌株的筛选及菌株生物特性研究  

Screening of High-yield Strain of Wuyiencin Gene wysR Overexpression and Its Biological Characteristics

作　　者：王家旺[1] 葛蓓孛[1] 刘艳[1] 刘彦彦[1] 张克诚[1] 

机构地区：[1]中国农业科学院植物保护研究所农业部作物有害生物综合治理重点实验室,北京100193

出　　处：《生物技术通报》2015年第8期125-131,共7页Biotechnology Bulletin

基　　金：公益性行业(农业)科研专项(201103002;201303025;国家自然科学基金项目(31371985;31401796)

摘　　要：以武夷菌素生物合成正调控基因wys R过表达菌株为初选材料,通过琼脂块法对2 060个过表达菌株单菌落进行初筛,得到35个优选菌株,再采用摇瓶发酵复筛的方法筛选出9个高产菌株,并通过菌株遗传稳定性分析,最终确定W-273菌株为最优菌株。研究武夷菌素wys R基因过表达菌株W-273的生理生化特征,发现与原始菌株CK-15相比,W-273菌株生长迅速,产孢量增加且产孢时间提前,W-273菌株生长3-4 d即可完成产孢,较菌株CK-15产孢时间提前了3-4 d;电子显微镜观察W-273菌株和CK-15菌株孢子形态不同,W-273菌株孢子呈椭圆形或杆状,CK-15菌株孢子呈圆形;通过摇瓶发酵表明W-273菌株较CK-15菌株代谢旺盛,抗生素产量提高79%-289%,且最佳发酵时间缩短4-8 h。In this research the over-expression strains with positive-regulating gene wysR synthesized from Wuyiencin were utilized as primary materials, 35 relatiVely optimal strains were selected from 2060 individual colonies of over-expression strains by agar blocks, then 9 high-yield strains were screened out adopting the method of shake flask fermentation. Ultimately, the strain W-273 was determined as the optimal strain by genetic stability analysis. Comparing the physiological and morphological characteristics between the strain of W-273 and original CK-15, W-273 strain grew faster, the production of spore was higher, and the time of spore production was in advance. The time to complete spore production for W-273 strain was 3-4 d, and this was in advance for 3-4 d than the original strain CK-15. Through electron microscope observation, the spores of W-273 were oval or rod, and the CK-15 strains were circular. Shake flask fermentation showed that strain W7273 had a higher metabolism compared to strain CK-15, antibiotic production increased 79%-289%, and the best fermentation time shortened 4-8 h.

关 键 词：武夷菌素 过表达菌株 wysR基因 筛选 

分 类 号：Q93[生物学—微生物学]