Abstract：Spinosad, produced by Saccharopolyspora spinosa, is a novel natural macrolide antibiotic with the properties of both safety of biopesticide and the efficiency of chemical pesticide. With an unique mechanism of action, spinosad has a weak toxicity towards mammalian, natural enemies of insects and environment. Thus, it has obtained permission and wildly used as a new biopesticide in different fields, such as pesticide, veterinary drug, sanitary field, etc. At present, the research of spinosad in our country is still in the laboratory scale due to its low yield. Industrial production of spinosad still has not been implemented for the lack of breeding of high-yield strains of spinosad. The mutation breeding systems and rational screening of Saccharopolyspora spinosa for high-yield spinosad were summarized in this paper. The common mutation types, including ultraviolet mutagenesis, ion beam implantation, γ ratial and atmospheric and room temperature plasma, have the features of high safety for the operators and environment, easy operation, rapid mutation capability, high mutation rate and genetic stability of targeted mutants. The optimization of the media, including seed and fermentation medium were reviewed. The effects of breeding temperature, dissolved oxygen, inoculum level, seeding age, medium volume, the initial pH and various precursors on the spinosad fermentation were also studied in the shaking flasks. Several precursors, such as short-chain alcohols, amino acids and methylating agents, could enhance biosynthesis of spinosad. The biosynthesis, separation and detection methods for this kind of compounds were also summarized. It is helpful for encouraging the industrial production of spinosad to develop high performance and accurate analysis methods. The domestic and foreign research status was analyzed from 4 aspects. Meanwhile, the trends in the development of spinosad were also discussed.
陈园. 2013. 多杀菌素高产菌株的诱变育种及快速筛选方法的研究[D]. 硕士学位论文, 华南理工大学, 导师：熊犍，pp. 18-23. (Chen Y. 2013. Mutation breeding of spinosad high-producing strain and study of rapid screening method[D]. Thesis for M.S., South China University of Technology, Supervisor: Xiong J, pp. 18-23.)陈继红，张利平，郭立格，等. 2008. 多杀菌素产生菌株航天育种效果研究[J]. 安徽农业科学, 36(12): 4951-4952. (Chen J H, Zhang L P, Guo L G, et al. 2008. Breening effect of space treatment breening on producting strains of spinosad[J]. Anhui Agriculture Science, 36(12): 4951-4952.) 兰周. 2012. 多杀菌素生物合成基因簇的克隆及异源表达载体的构建[D]. 硕士学位论文, 福建农林大学, 导师：关雄，pp. 18-26. （Lan Z. 2012. Cloning and construction of heterologoues expression vector of spinosad biosynthesis gene cluster[D]. Thesis for M.S., Fujian University of Agricultural and Forestry, Supervisor: Guan X, pp. 18-26.)梁艳. 2007. 多杀菌素高产菌株的选育和发酵条件的优化研究[D]. 硕士学位论文, 天津大学，导师：闻建平，pp. 17-31. (Liang Y. 2007. Study on the breeding of spinosad producing strain and optimization of fermentation conditions[D]. Thesis for M.S., Tianjin University , Supervisor: Wen J P, pp. 17-31.)黄颖，赵晨，杨博磊，等. 2014. 刺糖多孢菌高产菌株和野生型菌株多杀菌素生物合成基因簇(spn)在发酵过程中的表达比对[J]. 农业生物技术学报，11: 1337-1346. (Huang Y, Zhao C, Yang B L, et al. 2014. Expression Analysis of Spinosad Biosynthetic Gene Cluster(spn) in Hyperproducing and Wild-type Sacchar opolyspora spinosa strains During Fermentation[J]. Journal of Agricultural Biotechnology, 11: 1337-1346.)乔广军, 汪晨, 张凯, 等. 2012. 多杀菌素高产菌株的诱变选育[J]. 食品与发酵科技, 48(5): 30-34.(Qiao G J, Wang C, Zhang K, et al. 2012. Study on the breeding of spinosad producing strain[J]. Food Fermentaion Techniqual, 48(5): 30-34. )张雪，张晓菲，王立言, 等. 2014. 常压室温等离子体对微生物的作用机理及其应用基础研究[J]. 化工学报, 65(7): 2676-2684. (Zhang X, Zhang X F, Wang L Y, et al. 2014. Recent progress on atmospheric and room temperature plasma mutation breeding technology and its applications[J]. Journal of Chemical Technology, 65(7): 2676-2684.)王美玲，2013. 多杀菌素高产菌株的选育及发酵工艺的优化[D]. 硕士学位论文，天津大学，导师：卢玉文，pp: 14-22. (Wang M L, 2013. Mutation breeding of spinosad high-producing strain and study of rapid screening method[D]. Thesis for M.S., Tianjin University , Supervisor: Lu W Y, pp. 14-22.) 张苑. 2004. 多杀菌素高产菌种的推理选育和发酵工艺的优化[D]. 硕士学位论文, 浙江大学，导师：岑沛霖，pp. 26-32. (Zhang Y. 2004. Study on the breeding of spinosad producing strain and optimization of fermentation conditions[D]. Thesis for M.S., Zhejiang University, Supervisor: Cen P L, ) Boeck L D, Chio H, Eaton T E, et al. 1990. Macrolide compounds. Europe, EP375316[P]. Chen H L, Wan H G, Zhang J, et al. 2008. Mutation induction of pleurotus ferulae by low-energy N+ ion implantation and characters of the selected mutant[J]. Nuclear Science Techniques, 19(1): 13-16.Creemer, Kirst H A, Mynderse J S, et al. 1993. New A83543 compounds and process for production thereof, WO199309126A1[P].Guo W Q, Luo L S, LinN W, et al. 2012. Breeding high-yielding strains of spinosad by ion implantation[J]. Agriculure Science Technology, 14(4): 148-152.Hong L, Zhao Z, MelanconⅢ C E, et al. 2008. In vitro charac-terization of the enzymes involved in TDPD- forosamine biosynthesis in the spinosyn pathway of Saccharopolys-pora spinosa[J]. Journal of the American Chemical Society , 130(14): 4954-4967.Isiorho E A, Liu H, Keatinge Clay A T . 2012. Structural stud-ies of the spinosyn rhamnosyltransferase, SpnG[J]. Biochemistry, 51(6): 1213-1222.Jin Z H, Chen X, Cen P L. 2006. Effects of glucose and phosphate on spinosad fermentation by Saccharopolyspora spinosa[J]. Journal of Chinese Chemistry Engineer, 14(4): 542-546.Jin Z H, Xu B, Lin S Z, et al. 2009. Enhanced production of spinosad in Saccharopolyspora spinosa?by genome shuffling[J]. Application of Biochemistry and Biotechnology,?159(3):?655-663.Liang Y, Yu W Y,?Wen J P. 2009. Improvement of?Saccharopolyspora spinosa?and the kinetic analysis for spinosad production[J]. Application of Biochemistry and Biotechnology,?152(3): 440-448.Li J A, Han L M. 2008. An extraction method for spinosad, China, CN101560231A [P]. Mertz F P, Yao R C. 1990. Saccharopolyspora spinosasp. nov. isolated from soil collected in a sugar rum still[J]. International Journal of Systematic Bacteriology, 40 (1): 34-39.Myoyong L, David R W, Patricia N. 1999. Fluorescent excitation transfer immunoassay for the determination of spinosyn A in water[J]. Journal of Agriculture and Food Chemistry, 47: 2766-2770.Song W, Xiong J, Guo W Q, et al. 2009. Mutation effection of MNNG on strains of spinosad[J]. Chinese Journal of Biological Control, 25(2): 176-180.Strobel R J, Nakatsukasa W M. 1993. Response surface methods for optimizing Saccharopolyspora spinosa, a novel macrolide producer[J]. Journal of Industrial Microbiology Biotechnology, 11: 121-127.Ueno E, Oshima H,?Matsumoto H, et al. 2006. Determination?of?spinosad?in vegetables and fruits by high-performance liquid chromatography with UV and mass spectrometric detection after gel permeation chromatography and solid-phase?extraction?cleanup on a 2-layered column[J]. Journal of AOAC International, 89(6):?1641-1649.Wang C, Zhang X L, Chen Z, et al. 2009. Strain construction for enhanced production of spinosad via intergeneric protoplast fusion[J]. Canadian Journal of Microbiology, 55(9): 1070-1075.Waldron C, Madduri K, Crawford K. 2000. A cluster of genes for the biosynthesis of spinosyns, novel macrolide insect control agents produced by Saccharopolyspora spinosa[J]. Antonie Van Leeuwenhoek, 78(3): 385-390.Waldron C, Matsushima P, Rosteck P R, et al. 2001. Cloning and analysis of the spinosad biothetic gene cluster of Saccharopolyspora spinosa[J]. Chemistry and Biology, 8(5): 487-499.Wu Q, Li J G, Li D Z, et al. 2015. Determination residues of spinosad and spinetoram in soil by ultra performance liquid chromatography [J]. Chinese Journal of Pesticide Science, 17(1): 111-114.Xia L Q，Ding X Y, Yu Z Q, et al. 2010. An crafts for extraction of spinosad from the spinosyn fermentation broth, China, CN101906124B[P]. Xue L, Abd ElAty A M, Park J Y, et al. 2011. Determination of spinetoram in leafy vegetable crops using liquid chromatography and con?rmation via tandem mass spectrometry[J]. Chromatography, 25: 1099-1106.Zou X, Hang H F. 2009. Enhancement of erythromycin a production with feeding available nitrogen sources in erythromycin biosynthesis phase[J]. Bioresource Technology, 100: 3358-3365.Zhang S, Xia W, Zhu S R, et al. 2014. Fermentation Medium for Spinosad, China, CN103882079 [P].
