【作者】 张明政；

【导师】 林乃铨； 刘波；

【作者基本信息】 福建农林大学 ， 农药学， 2009， 硕士

【摘要】 本研究对生物农药苏云金芽孢杆菌Btlsz9408进行了气升式发酵,研究了发酵液的离心过滤工艺条件、壳聚糖絮凝浓缩工艺条件、复相乳液法制备Bt杀虫蛋白肠溶微胶囊的最佳工艺条件、羟丙基甲基纤维素邻苯二甲酸酯(HPMCP)蛋白质微胶囊包裹量的测定方法、杀虫蛋白的非肠溶性微胶囊、肠溶性微胶囊和非肠溶性微胶囊的释放特性等。气升式发酵得到了晶体规则,数量多的发酵原液。离心过滤可以除去发酵原液中的大颗粒物质,有利于后续分离纯化工艺的进行,其最佳工艺条件为在每次处理量1L,温度10-15℃、pH4.5时过滤效果最好。采用壳聚糖进行絮凝浓缩,避免了化学絮凝剂、微生物絮凝剂对原药效果的影响,絮凝浓缩可除去约75%左右的水分及有色物质,其最佳工艺条件为:pH5.0-6.0、温度30-35℃、壳聚糖浓度0.025%、搅拌时间5min、550r/min;通过对发酵液分离纯化,得到等电点沉淀法提取的杀虫蛋白质比有机溶剂法的纯度高。对测定HPMCP蛋白质微胶囊包裹量的方法研究中得到福林-酚法最适合测定。复相乳液法制备的肠溶式微胶囊形态多为圆形,并且没有粘结在一起的现象,最佳工艺条件为:分散质浓度为0.5%,搅拌转速6000r/min、囊壁材料浓度2%-3%、乳化剂用量为5%、芯材量4%、分散比为1:2.5-1:3.5。以乙基纤维素制备的非肠溶性微胶囊形态规则,不粘结在一起,通过对微胶囊释放特性研究得到:体外测定2-3h微胶囊释放量达到平衡,肠溶性微胶囊在偏碱性环境中释放速度快,而非肠溶性微胶囊则不受溶液的酸碱性影响。更多还原

【Abstract】 It studied air lift fermentation for Bt LSZ9408, the centrifugal filtration technology conditions of fermentation broth, the flocculation concentration technology condition of chitosan as flocculant for concentrating fermentation liquid. The optimization technology for preparation Bt pesticide protein to Enteric-coated microcapsules dosage form, the mensurating methods of protein envelopment quantum of hydroxy proprl methyl celluloxe phthalate protein microcapsules, un-enteric-coated microcapsules of Bt pesticide protein and the released character of enteric-coated microcapsules or not for Enteric-coated microcapsules were also studied in it.Many regular crystals of pesticide protein were obtained by air lift fermentation. The centrifugal filtration could eliminate big particles, which made for separation and purification of industry of Bt pesticide. The optimum technology condition was 1 L disposal quantity every time, temperature 10-15℃、pH 4.5. It could avoid the effect of chemical or microbial flocculant on active compound if chitosan as flocculant, which can remove about 75% water or coloured material, the optimal technology condition was pH5.0-6.0, temperature 30-35℃, chitosan concentration 0.025%, electromagnetic stirring time 5 min, electromagnetic stirring speed 550r/min. By separation fermentation liquid of Bt, which obtained that the pesticide protein purity extracted by isoelectric point precipitation method was higher than organic solvent method.Folin-pHenol assay was the appropriate method for detecting protein envelopment quantum of HPMCP microcapsules. The shape of enteric-coated microcapsules prepared by two pHase emulsion assay was rotundity and un-conglutinant, the optimum technology condition was dispersed pHase 0.5%, stirring speed 6000r/min, wall material 2%-3%, emulsifier concentration 5%, heartwood concentration 4%, dispersed proportion 1:2.5-1:3.5. The shape of un-enteric-coated microcapsules ethyl cellulose microcapsules was regular article and un-conglutinant. By the released character study of microcapsules which obtained that the released quantum reached peak value in two to three hours in vitro test, the enteric-coated released more quickly in alkalescence solution than in acidity solution but the un-enteric-coated was not.更多还原