高产酸性普鲁兰酶工程菌株种子强化工艺  

Seed Strengthening Process of Acid Pullulanase Producing Engineered Strain

作　　者：黄钦耿 林美花 黄勤勤 吴松刚[1,2] 黄建忠[1,2] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108 [2]工业微生物教育部工程研究中心,福建福州350108

出　　处：《精细化工》2017年第11期1239-1245,共7页Fine Chemicals

基　　金：国家高技术研究开发计划项目(No.2015AA021005)~~

摘　　要：为进一步提高酸性普鲁兰酶工程菌株(Baclicus subtilis F1147)的产酶水平,开展了工程菌株种子强化工艺的研究。采用小米为载体,制备菌株接触面大、通气量好的小米种培养基,小米种摇瓶发酵最佳接种量为0.33%,其发酵酶活较二代斜面种高64.5%,较摇瓶种高21.4%。30 L罐补料分批发酵,小米种最佳接种量为0.33%,种子罐周期约为8 h,较摇瓶种种子罐提前1 h,发酵最高酶活达1 209 U/m L,较摇瓶种罐高73.7%,生长及产酶周期较摇瓶种罐延长4 h,最高菌浓也较摇瓶种罐高出22.1%,显著促进菌株生长及延长产酶周期。并且小米种发酵有利于改善发酵液特性,提高陶瓷膜除菌效率,其除菌率达100%,滤液浓缩4倍,酶活力回收率可达90.4%,体现了较好的浓缩效果,为工程菌株的工业化放大应用奠定了良好基础。To further improve the yield of acid pullulanase produced by fermentation of the genetic engineering strain Baclicus subtilis F1147,the research on seed strengthening process was carried out.The millet medium with a large contact surface and good ventilation was prepared by using millet as a carrier.Under the condition of shaking flasks fermentation,the optimum inoculation amount of millet seed strain was 0.33%.The activity of pullulanase in millet seed flasks was 64.5% higher than that of the second generation slant,and 21.4% higher than that of shaking flasks seed.Subsequently,the fedbatch fermentation was carried out in a 30 L automatic fermenter.The seed culture cycle of millet（8 h）by seed strengthening process was reduced more efficiently than that of shaking flasks seed process（9 h）.The highest activity of pullulanase in seed strengthening process was 1 209 U/m L,which was73.7% higher than that of shaking flasks seed process.In addition,the growth and production cycle of pullulanase in millet strengthening process prolonged 4 hours compared with that of shaking flasks seed process,and the highest concentration of pullulanase was 22.1% higher than that of shaking flasks seed process,indicating that the seed strengthening process could significantly promote enzyme growth and extension of enzyme production cycle.In addition,seed enhancement was beneficial to improve the characteristics of fermentation broth and improve the bacteria removing efficiency of ceramic membrane,and the sterilization rate was up to 100%.The filtrate was concentrated 4 times,and the recovery rate of pullulanase activity was 90.4%,which laid a good foundation for the industrial application of genetic engineering strain Baclicus subtilis F1147.

关 键 词：普鲁兰酶 枯草芽孢杆菌 种子强化 补料分批发酵 膜处理 生物工程 

分 类 号：TQ426.97[化学工程]