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当前位置: 首页 > 文章 > 高产截短侧耳素产生菌高通量筛选方法的建立 中国兽医学报 2013,33 (10) 1590-1594
Position: Home > Articles > Identification of pleuromutilin-high-producing strains by high-throughput screening method Chinese Journal of Veterinary Science 2013,33 (10) 1590-1594

高产截短侧耳素产生菌高通量筛选方法的建立

作  者：
王学红;梁剑平;赵晓彬;刘宇;郝宝成
单  位：
农业部兽用药物创制重点实验室;中国农业科学院兰州畜牧与兽药研究所;甘肃省新兽药工程重点实验室
关键词：
截短侧耳素;高通量筛选;微生物育种
摘  要：
截短侧耳素类抗生素是一类副作用小、效果显著的抗菌化合物,其中真菌发酵产物截短侧耳素为人工合成各种截短侧耳素类抗生素的原料药,已大规模工业生产,市场需求巨大,急需选育高产菌株,利用高通量技术以提高筛选效率是可行的方法。通过固体发酵将发酵过程微量化于96孔板,恒温培养,确定高通量培养方法,截短侧耳素微孔板提取、显色,确定特征吸收波长,酶标仪测定特征波长吸收值计算截短侧耳素产量,确定高通量提取测定方法。结果表明,截短侧耳素经浓硫酸显色后,在455nm波长处有特征吸收峰,96孔板固体发酵和常规摇瓶发酵之间相关性良好,高产截短侧耳素产生菌的高通量筛选方法与常规方法相比具有快速准确的特性,此方法切实可行,结果可靠,大大提高截短侧耳素产生菌的筛选效率。
译  名：
Identification of pleuromutilin-high-producing strains by high-throughput screening method
作  者：
WANG Xue-hong;LIANG Jian-ping;ZHAO Xiao-bin;LIU Yu;HAO Bao-cheng;Key Laboratory of New Animal Drug Project,Gansu Province/Key Laboratory of Veterinary Pharmaceutics Dis covery,Ministry of Agriculture/Lanzhou Institute of Animal Science and Veterinary Pharma ceutics,Chinese Academy of Agricultural Sciences;
关键词：
pleuromutilin;;high-throughput screening;;microbial breeding
摘  要：
Pleuromutilin is one of the antibiotics that have few side effects,and its effect is significant.It is an active pharmaceutical ingredient,and has been industrially fermented in large-scale.The high-yield pleuromutilin strains were screened by the high-throughput screening.The solidstate fermentation miniaturize in 96-well microplates.Extract and color the pleuromutilin,according to the microplate reader UV absorption,measure the content of pleuromutilin.The characteristic absorption peak of colored pleuromutilin by concentrated H2SO4 is 455nm,the correlation between 96-well microplates and conventional fermentation is good,the high-throughput determination of extraction results is consistent with conventional detection methods,high-throughput screening method of high-yield pleuromutilin strain is faster and more accurate than the conventional method of screening,and has practical and reliable results,elevating the screening efficiency.

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