从芽胞杆菌dhs-330转座突变库筛选获得高产表面活性剂突变株  

Screening of a Bacillus sp. Mutant with High Surfactivity from dhs-330 Transposon Mutagenesis

作　　者：司晓光[1] 杜瑾[1] 郝建安[1] 姜天翔[1] 张晓青[1] 杨波[1] 张爱君[1] 张雨山[1] 王静[1] 

机构地区：[1]国家海洋局天津海水淡化与综合利用研究所,天津300192

出　　处：《生物技术通讯》2016年第6期790-793,共4页Letters in Biotechnology

基　　金：海洋公益性行业科研专项(201305022-5);中央级公益性科研院所基本科研业务费专项(K-JBYWF-2015-G17);中央级公益性科研院所基本科研业务费专项团队项目(K-JBYWF-2015-T11)

摘　　要：目的:利用本实验室构建的芽胞杆菌dhs-330的转座突变体库,筛选表面活性剂高产菌株,并获得最佳发酵条件。方法:通过排油圈法初筛和测定表面张力复筛,从500个突变株中筛选出5株表面活性较高的突变株,对获得的5株突变株进行遗传稳定性分析,通过单因素实验对高产表面活性剂的发酵条件进行优化。结果:获得了5株表面活性较高的突变株,传代培养10代后突变株dhs-330-021的遗传稳定性和表面活性均较好,dhs-330-021的最佳培养基为葡萄糖32 g/L、酵母提取物1.2 g/L、尿素5.0 g/L、KH_2PO_4 0.8 g/L、Na_2HPO_4·12H_2O 3 g/L、MgSO_4·7H_2O 0.15g/L、微量元素溶液1 m L/L,发酵液稀释至1/10后的表面张力降低到30.392 mN/m。结论:运用转座突变技术进行高产表面活性剂菌株的选育,可获得遗传稳定的高产菌株。Objective： To screen the mutants with high yield of surfactant from the Bacillus dhs-330 mutant li-brary in our laboratory, and to optimize the fermentation condition. Methods： By primary screening and re-screen-ing, 5 mutants with high surface activity were obtained from 500 mutants. The genetic stabilities of 5 mutantswere studied and the optimum fermentation conditions were determined by single-factor experiments. Results： Thegenetic stability and surface activity of dhs-330-021 were superior in 5 mutants after 10 generations. The opti-mum medium for dhs-330-021 was as follow： glucose 32 g/L, yeast extract 1.2 g/L, urea 5.0 g/L, KH_2PO_4 0.8 g/L,Na_2HPO_4·12H_2O 3 g/L, MgSO_4·7H_2O 0.15 g/L, trace elements 1 m L/L. The surface tension of fermentation brothdiluted 10-folds reached to 30.392 m N/m. Conclusion： It＇s an efficient path to obtain high-yield and genetic sta-bility strain by transposon mutagenesis technology.

关 键 词：表面活性剂 芽胞杆菌 转座突变 条件优化 突变株 

分 类 号：Q933[生物学—微生物学]