原生质体转化构建有机磷农药降解工程菌  

CONSTRUCTION OF ORGANOPHOSPHATE INSECTICIDES DEGRADING STRAIN BY PROTOPLAST TRANSFORMATION

作　　者：王永杰[1] 李顺鹏[1] 

机构地区：[1]南京农业大学微生物学系,南京210095

出　　处：《应用与环境生物学报》1999年第S1期162-165,共4页Chinese Journal of Applied and Environmental Biology

摘　　要：降解有机磷农药甲胺磷、敌敌畏和对硫磷的菌株地衣芽孢杆菌 Ｐ１２（ Ｂａｃｉｌｌｕｓｌｉｃｈｅｎｉｆｏｒｍｉｓ） 经溶菌酶处理获得原生质体，加入降解乐果的供体菌 Ｇ１ Ｄ Ｎ Ａ，经ρ（ Ｐ Ｅ Ｇ６ ０００） ＝ ３００ ｇ Ｌ－ １ 诱导，在液体再生培养基中振荡培养ｔ＝ ２０ ｈ ，使其恢复细胞壁后，离心收集菌体，涂布于含乐果的基础培养基上，经筛选得一转化子 Ｊ Ｐ Ｚ．其菌落形态明显不同于出发菌株，乐果平板连续传代１０ 次，性状保持稳定．在θ＝ ３０ ℃，１００ ｒ／ｍｉｎ 的培养条件下，３ ｄ 内对ρ（ 甲胺磷） ＝ ０ ．５ ｇ Ｌ－ １ ，ρ（ 敌敌畏） ＝ ０ ．２ ｇ Ｌ－ １ ，ρ（ 对硫磷） ＝ ０ ．１ ｇ Ｌ－ １ ，ρ（ 乐果） ＝ ０ ．３ ｇ Ｌ－ １ 的降解率分别为 Ｒｄ ＝ ７９ ．１ ％ ，４６ ．７ ％ ，２９ ．４ ％ 和４６ ．４ ％ ．The protoplasts of Bacillus licheniformis degrading methamidophos, dichlorvos and parathion were obtained by lysozyme treatment. Adding chromosomal DNA of the donator strain G1 and inducing with ρ (PEG 6 000)=300 g L -1 ,the cell walls were regenerated in the regeneration liquid medium by shaking. The cells were collected by centrifuge and sprayed on the basic medium containing dimethoate. The transformant J PZ were gained. The morphology of J PZ colonies was apparently different from that of the original strain. Its natures were stable when it was transferred for ten times in the plate containing dimethoate. Under the conditions of θ =30℃ and 100 r/min, the degradation rates of ρ (methamidophos)=0.5 g L -1 , ρ (dichlorvos)=0.2 g L -1 , ρ (paration) =0.1 g L -1 and ρ (dimethoate)=0.3 g L -1 were R d=79.1%,46.7%,29.4% and 46.4%, respectively in 3 d.

关 键 词：原生质体 转化 有机磷农药 降解 工程菌株 

分 类 号：X172[环境科学与工程—环境科学] Q813.2[生物学—生物工程]