麦芽糖转运相关基因的表达对刺糖多孢菌生长及多杀菌素合成的影响  

Effects of Expression of Maltose Transport Related Genes on Cell Growth and Spinosad Production in Saccharopolyspora spinosa

作　　者：黄颖[1,2] 赵晨[1] 张求学[1] 宋渊[2] 张晓琳[1] 

机构地区：[1]国家粮食局科学研究院,北京100037 [2]中国农业大学生物学院,北京100193

出　　处：《农业生物技术学报》2015年第11期1438-1444,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31300092);粮食公益性行业科研专项(No.201313002-3)

摘　　要：多杀菌素(spinosad)为土壤放线菌刺糖多孢菌(Saccharopolyspora spinosa)经有氧发酵产生的次级代谢产物,是一种广谱、高效、低毒的生物杀虫剂,已在农药、兽药和卫生用药领域得到广泛应用。刺糖多孢菌对长效碳源淀粉的利用能力很低,这给多杀菌素的工业化发酵带来了困难。本研究在刺糖多孢菌高产菌株ASAGF73中表达阿维链霉菌(Streptomyces avermitilis)的麦芽糖转运系统基因(maltose transporter gene)mal E、mal F和mal G,以提高菌株利用淀粉的能力。将整合型载体p SET152::mal EFG通过接合转移转化ASAGF73,获得的转化子成功表达了mal EFG基因。单一碳源发酵实验结果表明,转化子利用糊精和麦芽糖的能力显著提高。以糊精或麦芽糖为单一碳源的条件下,mal EFG的表达显著促进了菌体生物量的提高,进而提高多杀菌素的产量。本研究对于以淀粉为主要碳源发酵生产多杀菌素的工艺研究具有理论指导意义及应用价值。Spinosad, produced by Saccharopolyspora spinosa, is a kind of high-efficiency and low-toxicity bioinsecticide wildly used as pesticide, veterinary drug, hygienic insecticide, etc. The ability of S. spinosa to utilize starch and maltose was low, making a huge obstacle for the industrial fermentation of spinosad. In order to improve the ability of S. spinosa to use starch, the maltose transport related genes mal EFG from Streptomyces avermitilis were expressed in a spinosad high-yield strain S. spinosa ASAGF73. The integration expression vector p SET152：：mal EFG was transformed into ASAGF73 through conjugal transfer, and the mal EFG genes were successfully expressed in the resulting transformant. ASAGF73 and the transformant were cultured in single carbon source medium to study the effects of mal EFG, showing that the transformant consumed dextrin and maltose betterly than ASAGF73. In the media with dextrin or maltose as the sole carbon source, expression of mal EFG genes promoted obviously the growth of S. spinosa, resulting in improvement of spinosad yields. This study provides theoretical significance and application value for spinosad fermentation process using starch as main carbon source.

关 键 词：多杀菌素 刺糖多孢菌 麦芽糖转运系统 malEFG 基因表达 

分 类 号：S482.3[农业科学—农药学]