多杀菌素高产菌株的选育和发酵条件的优化研究
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摘要
多杀菌素是一种新型生物杀虫剂,具有以下优良特性:杀虫谱广,作用方式独特,杀虫活性高,半衰期短,残留低,无抗药性,对人畜无害,环境污染少等。多杀菌素在农牧业生产中具有广阔的应用前景,该产品的开发具有良好的经济效益和社会效益。多杀菌素的两种活性组分spinosyn A和spinosyn D,是刺糖多孢菌经有氧发酵产生的。目前,筛选高产菌株和优化发酵条件是提高多杀菌素产量的主要方法,也是国内外学者研究的热点。本文通过筛选多杀菌素高产菌株和优化发酵条件,使多杀菌素的产量提高了438.5%。
本论文根据多杀菌素的生物合成途径和代谢调节机理,对刺糖多孢菌进行理性化选育。以刺糖多孢菌原始菌株为出发菌株,经过紫外诱变,以2 mg/L鼠李糖和1.5%丙酸钠为筛选剂,筛选出抗鼠李糖和丙酸钠的多杀菌素高产突变株U-4213,通过前体正丙醇的添加实验,证明突变株对前体的耐受性增加,产素能力提高,产量达到120 mg/L,较出发菌株(产量为32.5 mg/L)提高了269.2%,且遗传性状稳定。
以突变株U-4213为出发菌株,通过一系列的单因子实验、Plackeet-Burman筛选实验和中心组合实验对培养基组分和发酵工艺条件进行优化,最终确定最佳的培养条件:葡萄糖68.5 g/L,麦芽糖10 g/L,棉籽蛋白26.0 g/L,玉米浆17.8 g/L,牛肉膏2.0 g/L,硫酸锌0.2 g/L,硫酸铵1.5 g/L,CaCO3 5 g/L,接种量10%,初始pH 7.0,摇床转速220 rpm,培养7天。在此条件下进行多杀菌素的发酵实验,得到多杀菌素的产量为175 mg/L,与优化前(120 mg/L)相比产量提高了45.8%。
最后建立了刺糖多孢菌原始菌株和突变菌株U-4213分批发酵培养的菌体生物量,多杀菌素浓度和底物消耗的动力学方程,方程的拟和值与实验值吻合很好,其相关系数(R2)分别为:0.9961,0.9955和0.9944以及0.9984,0.9934和0.9938,该结果表明,所选的动力学模型可以很好的描述多杀菌素的发酵行为,所建立的动力学方程能以高的准确度模拟刺糖多孢菌原始菌株发酵过程中的细胞生物量、产物多杀菌素浓度和底物浓度,为放大性实验研究提供基础。
Spinosad, a novel kind of biopesticide, has following excellent characteristics: effective on a wide variety of crops pest, unique mode of action, active to economically important pests, short half-life, low residue, non-drug resistant, large margins of safety to mammals, mankind and environmental profile. Spinosad will be widely used in agriculture and stockbreeding, and its exploitation has good economic and societal benefit. Spinosyn A and spinosyn D are the most active compositions of spinosad, which are producted by the aerobic fermentation of Saccharopolyspora spinosa. Nowadays, breeding high-producing strains and optimizing culture conditions are the research focus in the field of increasing the yield of spinosad. In this paper, the productivity of spinosad was improved by 438.5% through breeding high-producing strains and optimizing culture conditions.
A rational selection procedure was developed based on the biosynthetic pathway and metabolic regulation of spinosad. The wild type strain of Saccharopolyspora spinosa was treated with UV irradiation. A new mutant U-4213 was obtained using 2mg/L rhamnose and 1.5% sodium propionate as screening agents by resistant selection strategies in series. The results of experiment on tolerance to propyl alcohol showed that, in the mutant strain, the tolerance to precursor was improved and the level of producing spinosad was increased, the productivity of spinosad reached 120 mg/L, which was improved by 269.2% compared with that in the original strain (32.5 mg/L).
The optimization of the fermentation conditions for U-4213 was investigated by series single factor experiments, Plackeet-Burman selective experiment and central composite design experiment. The final obtained optimal condition was: glucose 68.5 g/L, maltose 10 g/L, cottonseed protein 26.0 g/L, corn steep liquor 17.8 g/L, beef extract 2.0 g/L, zinc sulfate 0.2 g/L, ammonium sulfate 1.5 g/L, calcium carbonate 5 g/L, the liquid volume 30 mL in 250 mL shaking flask, the inoculum ratio 10%, the initial pH 7.0,shaker speed 220 rpm, cultivate for 7 days. The productivity of spinosad under this optimal fermentation condition by shaking-flask batch fermentation reached 175 mg/L, which was increased by 45.8% compared with that under the initial fermentation condition (120 mg/L).
The kinetic models for biomass, substrate consumption and spinosad production were studied for the mutant strain U-4213 and the wild strain by conducting batch fermentation. The models performed well in simulating the cell, spinosad and carbohydrate concentrations of both wild strain and mutant strain with high accuracy (the correlative coefficient R2 were 0.9961, 0.9955, 0.9944 and 0.9984, 0.9934, 0.9938 respectively for wild strain and mutant strain). The result suggests that the kinetic models can describe well the process of spinosad fermentation, and will be a basic for the research of scaling up.
本论文根据多杀菌素的生物合成途径和代谢调节机理,对刺糖多孢菌进行理性化选育。以刺糖多孢菌原始菌株为出发菌株,经过紫外诱变,以2 mg/L鼠李糖和1.5%丙酸钠为筛选剂,筛选出抗鼠李糖和丙酸钠的多杀菌素高产突变株U-4213,通过前体正丙醇的添加实验,证明突变株对前体的耐受性增加,产素能力提高,产量达到120 mg/L,较出发菌株(产量为32.5 mg/L)提高了269.2%,且遗传性状稳定。
以突变株U-4213为出发菌株,通过一系列的单因子实验、Plackeet-Burman筛选实验和中心组合实验对培养基组分和发酵工艺条件进行优化,最终确定最佳的培养条件:葡萄糖68.5 g/L,麦芽糖10 g/L,棉籽蛋白26.0 g/L,玉米浆17.8 g/L,牛肉膏2.0 g/L,硫酸锌0.2 g/L,硫酸铵1.5 g/L,CaCO3 5 g/L,接种量10%,初始pH 7.0,摇床转速220 rpm,培养7天。在此条件下进行多杀菌素的发酵实验,得到多杀菌素的产量为175 mg/L,与优化前(120 mg/L)相比产量提高了45.8%。
最后建立了刺糖多孢菌原始菌株和突变菌株U-4213分批发酵培养的菌体生物量,多杀菌素浓度和底物消耗的动力学方程,方程的拟和值与实验值吻合很好,其相关系数(R2)分别为:0.9961,0.9955和0.9944以及0.9984,0.9934和0.9938,该结果表明,所选的动力学模型可以很好的描述多杀菌素的发酵行为,所建立的动力学方程能以高的准确度模拟刺糖多孢菌原始菌株发酵过程中的细胞生物量、产物多杀菌素浓度和底物浓度,为放大性实验研究提供基础。
Spinosad, a novel kind of biopesticide, has following excellent characteristics: effective on a wide variety of crops pest, unique mode of action, active to economically important pests, short half-life, low residue, non-drug resistant, large margins of safety to mammals, mankind and environmental profile. Spinosad will be widely used in agriculture and stockbreeding, and its exploitation has good economic and societal benefit. Spinosyn A and spinosyn D are the most active compositions of spinosad, which are producted by the aerobic fermentation of Saccharopolyspora spinosa. Nowadays, breeding high-producing strains and optimizing culture conditions are the research focus in the field of increasing the yield of spinosad. In this paper, the productivity of spinosad was improved by 438.5% through breeding high-producing strains and optimizing culture conditions.
A rational selection procedure was developed based on the biosynthetic pathway and metabolic regulation of spinosad. The wild type strain of Saccharopolyspora spinosa was treated with UV irradiation. A new mutant U-4213 was obtained using 2mg/L rhamnose and 1.5% sodium propionate as screening agents by resistant selection strategies in series. The results of experiment on tolerance to propyl alcohol showed that, in the mutant strain, the tolerance to precursor was improved and the level of producing spinosad was increased, the productivity of spinosad reached 120 mg/L, which was improved by 269.2% compared with that in the original strain (32.5 mg/L).
The optimization of the fermentation conditions for U-4213 was investigated by series single factor experiments, Plackeet-Burman selective experiment and central composite design experiment. The final obtained optimal condition was: glucose 68.5 g/L, maltose 10 g/L, cottonseed protein 26.0 g/L, corn steep liquor 17.8 g/L, beef extract 2.0 g/L, zinc sulfate 0.2 g/L, ammonium sulfate 1.5 g/L, calcium carbonate 5 g/L, the liquid volume 30 mL in 250 mL shaking flask, the inoculum ratio 10%, the initial pH 7.0,shaker speed 220 rpm, cultivate for 7 days. The productivity of spinosad under this optimal fermentation condition by shaking-flask batch fermentation reached 175 mg/L, which was increased by 45.8% compared with that under the initial fermentation condition (120 mg/L).
The kinetic models for biomass, substrate consumption and spinosad production were studied for the mutant strain U-4213 and the wild strain by conducting batch fermentation. The models performed well in simulating the cell, spinosad and carbohydrate concentrations of both wild strain and mutant strain with high accuracy (the correlative coefficient R2 were 0.9961, 0.9955, 0.9944 and 0.9984, 0.9934, 0.9938 respectively for wild strain and mutant strain). The result suggests that the kinetic models can describe well the process of spinosad fermentation, and will be a basic for the research of scaling up.
引文
[1]唐除痴，农药化学，天津：南开大学出版社，1998
[2]张兴，马志卿，李广泽，试谈生物农药的定义和范畴，农药科学与管理，2002，23(1)：32—36
[3]张兴，马志卿，李广泽等，生物农药评述，西安农林科技大学学报(自然科学版)，2002，30(2)：142—147
[4]高菊芳，生物农药的作用、应用与功效(一)，世界农药，2001，23(1)：1—6
[5]Pillmoor J B，Wright K，TerryA S，Natural products as a source of agochemicalsand leads for chemical synthesis，Pesticide Science，1993，39：131—140
[6]吴霞，多杀菌素-以天然产物开发新农药的范例，世界农药，23(1)：24—28
[7]Boeck L Dwaine，Chio H，Eaton T E，et al，Eur．Pat．Appl．EP 375316，1990
[8]Kirst H，Michel K，Discovery，isolation and structure elucidation of a family ofstructurally unique fermentation—derived tetracyclic macrolides，Baker D，FenyesJ，Synthesis and Chemistry of Agrochemicals III，Washington，Dc：AmericanChemical Society，1992，214—225
[9]Thompson G D，Michel K H，Yao R C，et al，The discovery ofSaccharopolysporaspinosa and a new class of insect control products，Down to Earth，1997，52(1)：1—5
[10]Kirst H A，Michel K H，Martin J，et al，A83543A—D，unique fermentation—derivedtetracyclic macrolides，Tetrahedron Letters，1991，32(37)：4839—4842
[11]Turner J R，Mary L B，Broughton M C，Process for the production of A83543compounds with Saccharopolyspora spinosa，United States Patent 5591606，1997
[12]Baker P J，Process for isolating A83543 and its components，United States Patent5227295，1993
[13]Waldron C，Madduri K，Crawford K，A cluster of genes for the biosynthesis ofspinosyns，novel macrolide insect control agents produced by Saccharopolysporaspinosa，Antonie Van Leeuwenhoek，2000，78(324)：385—390
[14]张部昌，赵志虎，马清钧，红霉素生物合成的分子生物学，生物技术通讯，2001，12(2)：151—160
[15]Ikeda H，Nonomiya T，Usami M，et al，Organization of the biosynthetic genecluster for the polyketide anthelmintic macrolide avermectin in Streptomycesavermitils，Proceedings of the National Academy of Science，1999，96：9509—9514
[16]郝卫民，弗氏链霉菌基因组的tylIBA区域五个泰洛星生物合成基因的分析，国外医药抗生素分册，1996，17(6)：401--403
[17]李荣贵，王普，梅建凤等，新型生物杀虫剂-刺糖菌素，微生物学通报，2003，30(1)：77—92
[18]Krishnamurthy Madduri，Clive Waldron，Donald J Merlo，Rhamnosebiosynthesis pathway supplies precursors for primary and secondary metabolismin Saccharopolyspora spinosa，Journal of Bacteriology，2001，183(19)：5632—5638
[19]Clive Waldron，Patti Matsushima，Paul R，et al，Cloning and analysis of thespinosad biosynthetic gene cluster of Saccharopolyspora spinosa，ChemistryBiology．2001．8：487—499
[20]Mynderse，A83543 compounds and process for production thereof,United StatesPatent 5670486，1997
[21]Baker P J，Process for producing A83543 compounds，United States Patent5362634，1994
[22]Sparks T C，Biological activity of the spinosyns，fermentation derived insectcontrol agents on tabacco budworm larvae，Journal of Economic Entomology，1988，91：1227—1283
[23]Salgado V L，Studies on the mode of action of Spinosad：Insect Symptoms andPhysiological Correlates，Pesticide Biochemistry and Physiology，1998，60(2)：91—102
[24]Salgado V L，Sheers J J，Watson G B，et al，Studies on the mode of action ofspinosad：the internal effective concentration and the concentration dependenceon neural excitation，Pesticide Biochemistry and Physiology，1998，60(2)：103—110
[25]Liu N，Yue X，Insecticide resistance and cross—resistance in the housefly(Diptera：Muscidae)，Journal of Economic Entomology，2000，93(4)：1269—1275
[26]Carson W G Trumhle J T Effect of insecticides on celervinsects，ArthropodManagement Tests．1997．22：117
[27]Palurnbo J C，Evaluation of selective insecticides for control of lepidopterouslarvae in lettuce，Arthropod Management Tests，1997，22：136
[28]Riley D G Evaluation of insecticide treatments on cabbage，ArthropodManagement Tests，1998，23：82
[29]Schuster D J，Management of insects on flesh market tomatoes，spring，1996，Arthropod Management Tests．1997．22：182
[30]Webb S E，Control of pickleworm on squash with selective insecticides，1997，Arthropod Management Tests，1998，23：142—143
[31]潘登明，马艳，贾海庆，48％Spinosad浓溶剂防治棉铃虫试验研究，中国棉花，2001，28(1)：15—16
[32]林文彩，郭世俭，章金明等，菜喜和集琦虫蜘克对小菜蛾生长发育、存活和生殖的影响，浙江农业学报，2001，13(3)：152—156
[33]张友军，王光锋，吴青君等，多杀菌素对不同发育阶段甜菜夜蛾的毒力及其体内超氧化物歧化醛、过氧化氢酶和过氧化物酶的影响，农药学学报，2003，5(3)：31--38
[34]Thompson G D，Dutton R，Sparks T C，Spinosad a case study：an example from anatural products discovery programme，Pest Management Science，2000，56(8)：696—702
[35]Carson W Trumble J，Effect of insecticides for control of leafminers onlimabeans，1997，Arthropod Management Tests，1998，23(1)：74—75
[36]Kerns D，Control of lepidopterous larvae and leafminers in lettuce，1995，Arthropod Management Tests，1996，21(2)：117—118
[37]Linduska J，Foliarsprays to control earinvading insects on sweet corn，1 997，Arthropod Management Tests，1998，23(1)：95—96
[38]SaundersD，Fate of spinosad in the enviroment，Down to EaCh，1996，52(1)：21—28
[39]Gary D，Robert D，Spinosad a case study：an example from a nature productsdiscovery programme，Pest Managament Science，2000，56(8)：696—702
[40]Schoonover J，Larson L，Laboratory activity of spinosad on nontarget beneficialarthropods，Arthropod Management Tests，1995，20(5)：357．
[41]Nayak M K，Effectiveness of spinosad as a grain protectant against resistanbeetle and peocid pests of stored grain in Australia，Journal of Stored ProductsResearch，41(4)：455—467
[42]Pineda S，Effects of two biorational insecticides，spinosad andmethoxyfenozide，on Spodoptera littoralis (Lepidoptera：Noctuidse) underlaboratory conditions，Journal ofEconomic Entomology，97(6)：1906—1911
[43]Jones T The efficacy of spinosad against the western flower thrips，Frankliniellaoccidentalis，and its impact on associated biological control agents on greenhousecucumbers in southern Ontario，Pest Management Science，61(2)：179—185
[44]Downard P, Spinosad controls a range of lepidoptern pests in crucifers inAustralia，Management of Dismondback moth and Other Crucifer Pests，351-355
[45]Herron G A，James T M，2005，Monitoring insecticide resistance in AustralianFrankliniella occidentalis Pergande(Thysanoptera：Thripidate)detects fipronil andspinosad resistance，Australian Journal ofEntomology，44：299—303
[46]Shono T Scott J G Spinosad resistance in the housefly，Musca domestica，iS dueto a recessive factor on autosome 1，Pesticide Biochemistry and Physiology, 2003，75(1—2)：17
[47]唐振华，庄佩君，多杀菌素A／D及甜菜夜蛾对该药剂的抗药性，世界农药，2001，23(3)：38--40
[48]Gary D，Crouse T C，Sparks C V et al，Chemistry and insecticide activity of thespinosyns，The Food—environment Change，1998，56：158—160
[49]Creemer L C，Kirst H A，Paschal J W et al，Synthesis and insecticidal activity ofspinosyn Analogs functionally altered at the 2’-．3’-and 4’-positions of therhamnose moiety，Journal of Antibiotic，2000，53：171—178
[50]Crouse G D，Sparks T C，Naturally derived materials as products and leads forinsect control：the spinosyns，Reviews of Toxicology，1998，2：133—146
[51]Crouse G D，Sparks T C，Schoonover J，et al，Recent advances in the chemistryof spinosyns，Pest Management Science，2001，57：177—185
[52]Sparks T C，Anzeveno P B，Martynow J G，et al，The application of artificialneural networks to the identification of new spinosoids with improved biologicalactivity toward larvae of Heliothis virescens，Pesticide Biochemistry andPhysiology，2000，67：187—197
[53]Sparks T C，Crouse G D，Durst G Natural products as insecticides：the biology，biochemistry and quantitative structure—activity relationships of spinosyns andspinosoids，Pest Management Science，2001，57：896—905
[54]Jeschke，Method for producing novel spinosyn derivatives，United States Patent7034130，2006
[55]F P Mertz，R C Yao，Saccharopolyspora spinosa sp．nov．Isolated from soilcollected in a sugar mill rum still，International Journal of System Bacteriology，1990，40：34—39
[56]贺玉平，戴经元，代鹏等，多杀菌素高产菌株的选育．中国生物防治，22(1)：37—39
[57]苏建亚，沈晋良，多杀菌素的生物合成，中国生物工程杂志，23(5)：55—59
[58]Mynderse，A83543 compounds and process for production thereof,United StatesPatent 5840861，1998
[59]Mynderse，A83543 compounds and process for production thereof,United StatesPatent 5670364，1997
[60]张苑，金志华，林建平等，多杀菌素的高效液相色谱法测定，农药，2003，42(10)：27—28
[61]Lee M，Walt D R，Nutent P J，Agricultural Food Chemistry，1999：47：2766—2770
[62]吴红宇，多杀菌素(Spinosads)产生菌Saccharopolyspora spinosa产素能力的提高及发酵工艺的初步研究：[硕士学位论文]，北京；中国农业大学，2004
[63]代朋，多杀菌素生产菌株育种及发酵工艺研究：[硕士学位论文]，武汉；华中农业大学，2004
[64]R J Strobel，Jr,W M Nakatsukasa，Response surface methods for optimizingSaccharopolyspora spinosa，a novel macrolide producer,Journal of IndustrialMicrobiology，1993，11：121—127
[65]Krishnan S，Prapulla S G Raj alakshmi D，et al，Screening and selection of mediacomponents for lactic acid production using Plackett—Burman design，BioprocessEngineering，1998，19：61—65
[66]Ahuja S K，Ferreira G M，Moreira A R，Application of Plackett—Burman designand response surface methodology to achieve exponential growth for aggregatedshipworm bacterium，Biotechnology and Bioengineering，2004，85(6)：666—675
[67]Plackett R L，Burman J P, The design of optimum multifactorial experiments，Biometrika，1946，33：305—325
[68]马小魁，陈五岭，陈勇，利用Minitab软件优化酵母培养基，食品与发酵工业，2005，31(12)：72—75
[69]Goudar C T，Joeris K，Logistic equations effectively model mammalian cellbatch and fed—batch kinetics by logically constraining the fit，BiotechnologyProgress，2005，21(4)：1109—1118
[70]Luedeking R，Piret E L，A kinetic study of the lactic acid fermentation：Batchprocess at controlled pH，Journal of Biochemical MicrobiologicalBiotechnolofgical Engineering，1959，1：363—394
[71]张竞立，钱曙光，廖美德等，刺糖多孢菌分批发酵动力学研究，华南农业大学学报，2006，27(3)：51--54
[72]Hesketh A，K ochi，A novel method for improving Streptomyces coelicolor A3(2)for production of actinorhodin by introduction of rpsL(Encoding ribosomalprotein S12)mutation conferring resistance to streptomycin，Journal of Antibiotic，1997，50：532
[73]JIN Zhi—hua，WU Jian—ping，ZHANG Yuan，et al，Improvement of spinosadproducing Saccharopolyspora spinosa by rational screening，Journal of Zhej iangUniversity SCIENCE A，2006，7：366—370
[74]Sergio Sanchez，Arnold L Demain，Metabolic regulation of fermentationprocesses，Enzyme and Microbial Technology，2002，31：895—906
[75]王菊芳，吴晖，前体物质对阿维菌素生物合成的影响，华南理工大学学报(自然科学版)，2002，30(5)：16—18
[76]顾觉奋，郎天戈，丙酸盐对大环内酯类抗生素M-90生物合成的调控，中国抗生素杂志，1998，23(2)：136—139
[77]张蔚文，焦瑞身，丙酸盐对利福霉素生物合成的调控，中国抗生素杂志，1995，20(4): 268—272
[78]刘峰，陈浦云，陈详明，红霉素高产变株F1—57的选育及发酵工艺研究，福建师范大学学报(自然科学版)，2003，19(2)：61--64
[79]梁桂兆，卞科，基于响应面分析法的可食性小麦蛋白膜的研究，食品科学，2004，25(5):70—76
[80]韩永龙，王浩，付丽佳，SPSS软件用于药学研究中的正交设计数据处理，药学进展，2003，26(3)：179—182
[81]侯学文，悬浮玫瑰茄细胞的生长及泛醌合成的代谢调控，广州：华南理工大学食品与生物工程学院，1998
[82]Martin J F，McDaniel L E，Kinetics of biosynthesis of polyene macrolideantibiotics in batch cultures：cell maturation time，Biotechnology andBiOengineering，1975，17：925—38
[2]张兴，马志卿，李广泽，试谈生物农药的定义和范畴，农药科学与管理，2002，23(1)：32—36
[3]张兴，马志卿，李广泽等，生物农药评述，西安农林科技大学学报(自然科学版)，2002，30(2)：142—147
[4]高菊芳，生物农药的作用、应用与功效(一)，世界农药，2001，23(1)：1—6
[5]Pillmoor J B，Wright K，TerryA S，Natural products as a source of agochemicalsand leads for chemical synthesis，Pesticide Science，1993，39：131—140
[6]吴霞，多杀菌素-以天然产物开发新农药的范例，世界农药，23(1)：24—28
[7]Boeck L Dwaine，Chio H，Eaton T E，et al，Eur．Pat．Appl．EP 375316，1990
[8]Kirst H，Michel K，Discovery，isolation and structure elucidation of a family ofstructurally unique fermentation—derived tetracyclic macrolides，Baker D，FenyesJ，Synthesis and Chemistry of Agrochemicals III，Washington，Dc：AmericanChemical Society，1992，214—225
[9]Thompson G D，Michel K H，Yao R C，et al，The discovery ofSaccharopolysporaspinosa and a new class of insect control products，Down to Earth，1997，52(1)：1—5
[10]Kirst H A，Michel K H，Martin J，et al，A83543A—D，unique fermentation—derivedtetracyclic macrolides，Tetrahedron Letters，1991，32(37)：4839—4842
[11]Turner J R，Mary L B，Broughton M C，Process for the production of A83543compounds with Saccharopolyspora spinosa，United States Patent 5591606，1997
[12]Baker P J，Process for isolating A83543 and its components，United States Patent5227295，1993
[13]Waldron C，Madduri K，Crawford K，A cluster of genes for the biosynthesis ofspinosyns，novel macrolide insect control agents produced by Saccharopolysporaspinosa，Antonie Van Leeuwenhoek，2000，78(324)：385—390
[14]张部昌，赵志虎，马清钧，红霉素生物合成的分子生物学，生物技术通讯，2001，12(2)：151—160
[15]Ikeda H，Nonomiya T，Usami M，et al，Organization of the biosynthetic genecluster for the polyketide anthelmintic macrolide avermectin in Streptomycesavermitils，Proceedings of the National Academy of Science，1999，96：9509—9514
[16]郝卫民，弗氏链霉菌基因组的tylIBA区域五个泰洛星生物合成基因的分析，国外医药抗生素分册，1996，17(6)：401--403
[17]李荣贵，王普，梅建凤等，新型生物杀虫剂-刺糖菌素，微生物学通报，2003，30(1)：77—92
[18]Krishnamurthy Madduri，Clive Waldron，Donald J Merlo，Rhamnosebiosynthesis pathway supplies precursors for primary and secondary metabolismin Saccharopolyspora spinosa，Journal of Bacteriology，2001，183(19)：5632—5638
[19]Clive Waldron，Patti Matsushima，Paul R，et al，Cloning and analysis of thespinosad biosynthetic gene cluster of Saccharopolyspora spinosa，ChemistryBiology．2001．8：487—499
[20]Mynderse，A83543 compounds and process for production thereof,United StatesPatent 5670486，1997
[21]Baker P J，Process for producing A83543 compounds，United States Patent5362634，1994
[22]Sparks T C，Biological activity of the spinosyns，fermentation derived insectcontrol agents on tabacco budworm larvae，Journal of Economic Entomology，1988，91：1227—1283
[23]Salgado V L，Studies on the mode of action of Spinosad：Insect Symptoms andPhysiological Correlates，Pesticide Biochemistry and Physiology，1998，60(2)：91—102
[24]Salgado V L，Sheers J J，Watson G B，et al，Studies on the mode of action ofspinosad：the internal effective concentration and the concentration dependenceon neural excitation，Pesticide Biochemistry and Physiology，1998，60(2)：103—110
[25]Liu N，Yue X，Insecticide resistance and cross—resistance in the housefly(Diptera：Muscidae)，Journal of Economic Entomology，2000，93(4)：1269—1275
[26]Carson W G Trumhle J T Effect of insecticides on celervinsects，ArthropodManagement Tests．1997．22：117
[27]Palurnbo J C，Evaluation of selective insecticides for control of lepidopterouslarvae in lettuce，Arthropod Management Tests，1997，22：136
[28]Riley D G Evaluation of insecticide treatments on cabbage，ArthropodManagement Tests，1998，23：82
[29]Schuster D J，Management of insects on flesh market tomatoes，spring，1996，Arthropod Management Tests．1997．22：182
[30]Webb S E，Control of pickleworm on squash with selective insecticides，1997，Arthropod Management Tests，1998，23：142—143
[31]潘登明，马艳，贾海庆，48％Spinosad浓溶剂防治棉铃虫试验研究，中国棉花，2001，28(1)：15—16
[32]林文彩，郭世俭，章金明等，菜喜和集琦虫蜘克对小菜蛾生长发育、存活和生殖的影响，浙江农业学报，2001，13(3)：152—156
[33]张友军，王光锋，吴青君等，多杀菌素对不同发育阶段甜菜夜蛾的毒力及其体内超氧化物歧化醛、过氧化氢酶和过氧化物酶的影响，农药学学报，2003，5(3)：31--38
[34]Thompson G D，Dutton R，Sparks T C，Spinosad a case study：an example from anatural products discovery programme，Pest Management Science，2000，56(8)：696—702
[35]Carson W Trumble J，Effect of insecticides for control of leafminers onlimabeans，1997，Arthropod Management Tests，1998，23(1)：74—75
[36]Kerns D，Control of lepidopterous larvae and leafminers in lettuce，1995，Arthropod Management Tests，1996，21(2)：117—118
[37]Linduska J，Foliarsprays to control earinvading insects on sweet corn，1 997，Arthropod Management Tests，1998，23(1)：95—96
[38]SaundersD，Fate of spinosad in the enviroment，Down to EaCh，1996，52(1)：21—28
[39]Gary D，Robert D，Spinosad a case study：an example from a nature productsdiscovery programme，Pest Managament Science，2000，56(8)：696—702
[40]Schoonover J，Larson L，Laboratory activity of spinosad on nontarget beneficialarthropods，Arthropod Management Tests，1995，20(5)：357．
[41]Nayak M K，Effectiveness of spinosad as a grain protectant against resistanbeetle and peocid pests of stored grain in Australia，Journal of Stored ProductsResearch，41(4)：455—467
[42]Pineda S，Effects of two biorational insecticides，spinosad andmethoxyfenozide，on Spodoptera littoralis (Lepidoptera：Noctuidse) underlaboratory conditions，Journal ofEconomic Entomology，97(6)：1906—1911
[43]Jones T The efficacy of spinosad against the western flower thrips，Frankliniellaoccidentalis，and its impact on associated biological control agents on greenhousecucumbers in southern Ontario，Pest Management Science，61(2)：179—185
[44]Downard P, Spinosad controls a range of lepidoptern pests in crucifers inAustralia，Management of Dismondback moth and Other Crucifer Pests，351-355
[45]Herron G A，James T M，2005，Monitoring insecticide resistance in AustralianFrankliniella occidentalis Pergande(Thysanoptera：Thripidate)detects fipronil andspinosad resistance，Australian Journal ofEntomology，44：299—303
[46]Shono T Scott J G Spinosad resistance in the housefly，Musca domestica，iS dueto a recessive factor on autosome 1，Pesticide Biochemistry and Physiology, 2003，75(1—2)：17
[47]唐振华，庄佩君，多杀菌素A／D及甜菜夜蛾对该药剂的抗药性，世界农药，2001，23(3)：38--40
[48]Gary D，Crouse T C，Sparks C V et al，Chemistry and insecticide activity of thespinosyns，The Food—environment Change，1998，56：158—160
[49]Creemer L C，Kirst H A，Paschal J W et al，Synthesis and insecticidal activity ofspinosyn Analogs functionally altered at the 2’-．3’-and 4’-positions of therhamnose moiety，Journal of Antibiotic，2000，53：171—178
[50]Crouse G D，Sparks T C，Naturally derived materials as products and leads forinsect control：the spinosyns，Reviews of Toxicology，1998，2：133—146
[51]Crouse G D，Sparks T C，Schoonover J，et al，Recent advances in the chemistryof spinosyns，Pest Management Science，2001，57：177—185
[52]Sparks T C，Anzeveno P B，Martynow J G，et al，The application of artificialneural networks to the identification of new spinosoids with improved biologicalactivity toward larvae of Heliothis virescens，Pesticide Biochemistry andPhysiology，2000，67：187—197
[53]Sparks T C，Crouse G D，Durst G Natural products as insecticides：the biology，biochemistry and quantitative structure—activity relationships of spinosyns andspinosoids，Pest Management Science，2001，57：896—905
[54]Jeschke，Method for producing novel spinosyn derivatives，United States Patent7034130，2006
[55]F P Mertz，R C Yao，Saccharopolyspora spinosa sp．nov．Isolated from soilcollected in a sugar mill rum still，International Journal of System Bacteriology，1990，40：34—39
[56]贺玉平，戴经元，代鹏等，多杀菌素高产菌株的选育．中国生物防治，22(1)：37—39
[57]苏建亚，沈晋良，多杀菌素的生物合成，中国生物工程杂志，23(5)：55—59
[58]Mynderse，A83543 compounds and process for production thereof,United StatesPatent 5840861，1998
[59]Mynderse，A83543 compounds and process for production thereof,United StatesPatent 5670364，1997
[60]张苑，金志华，林建平等，多杀菌素的高效液相色谱法测定，农药，2003，42(10)：27—28
[61]Lee M，Walt D R，Nutent P J，Agricultural Food Chemistry，1999：47：2766—2770
[62]吴红宇，多杀菌素(Spinosads)产生菌Saccharopolyspora spinosa产素能力的提高及发酵工艺的初步研究：[硕士学位论文]，北京；中国农业大学，2004
[63]代朋，多杀菌素生产菌株育种及发酵工艺研究：[硕士学位论文]，武汉；华中农业大学，2004
[64]R J Strobel，Jr,W M Nakatsukasa，Response surface methods for optimizingSaccharopolyspora spinosa，a novel macrolide producer,Journal of IndustrialMicrobiology，1993，11：121—127
[65]Krishnan S，Prapulla S G Raj alakshmi D，et al，Screening and selection of mediacomponents for lactic acid production using Plackett—Burman design，BioprocessEngineering，1998，19：61—65
[66]Ahuja S K，Ferreira G M，Moreira A R，Application of Plackett—Burman designand response surface methodology to achieve exponential growth for aggregatedshipworm bacterium，Biotechnology and Bioengineering，2004，85(6)：666—675
[67]Plackett R L，Burman J P, The design of optimum multifactorial experiments，Biometrika，1946，33：305—325
[68]马小魁，陈五岭，陈勇，利用Minitab软件优化酵母培养基，食品与发酵工业，2005，31(12)：72—75
[69]Goudar C T，Joeris K，Logistic equations effectively model mammalian cellbatch and fed—batch kinetics by logically constraining the fit，BiotechnologyProgress，2005，21(4)：1109—1118
[70]Luedeking R，Piret E L，A kinetic study of the lactic acid fermentation：Batchprocess at controlled pH，Journal of Biochemical MicrobiologicalBiotechnolofgical Engineering，1959，1：363—394
[71]张竞立，钱曙光，廖美德等，刺糖多孢菌分批发酵动力学研究，华南农业大学学报，2006，27(3)：51--54
[72]Hesketh A，K ochi，A novel method for improving Streptomyces coelicolor A3(2)for production of actinorhodin by introduction of rpsL(Encoding ribosomalprotein S12)mutation conferring resistance to streptomycin，Journal of Antibiotic，1997，50：532
[73]JIN Zhi—hua，WU Jian—ping，ZHANG Yuan，et al，Improvement of spinosadproducing Saccharopolyspora spinosa by rational screening，Journal of Zhej iangUniversity SCIENCE A，2006，7：366—370
[74]Sergio Sanchez，Arnold L Demain，Metabolic regulation of fermentationprocesses，Enzyme and Microbial Technology，2002，31：895—906
[75]王菊芳，吴晖，前体物质对阿维菌素生物合成的影响，华南理工大学学报(自然科学版)，2002，30(5)：16—18
[76]顾觉奋，郎天戈，丙酸盐对大环内酯类抗生素M-90生物合成的调控，中国抗生素杂志，1998，23(2)：136—139
[77]张蔚文，焦瑞身，丙酸盐对利福霉素生物合成的调控，中国抗生素杂志，1995，20(4): 268—272
[78]刘峰，陈浦云，陈详明，红霉素高产变株F1—57的选育及发酵工艺研究，福建师范大学学报(自然科学版)，2003，19(2)：61--64
[79]梁桂兆，卞科，基于响应面分析法的可食性小麦蛋白膜的研究，食品科学，2004，25(5):70—76
[80]韩永龙，王浩，付丽佳，SPSS软件用于药学研究中的正交设计数据处理，药学进展，2003，26(3)：179—182
[81]侯学文，悬浮玫瑰茄细胞的生长及泛醌合成的代谢调控，广州：华南理工大学食品与生物工程学院，1998
[82]Martin J F，McDaniel L E，Kinetics of biosynthesis of polyene macrolideantibiotics in batch cultures：cell maturation time，Biotechnology andBiOengineering，1975，17：925—38
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