辅酶Q_(10)产生菌的原生质体制备与再生条件研究  

Study on preparation and regeneration of protoplasts from CoQ_(10)-yield strains

作　　者：王飞飞[1] 岳田利[1] 袁亚宏[1] 高振鹏[1] 

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2009年第3期208-212,218,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"十一五"科技支撑计划项目(2006BAK02A18;2006BAK02A24;2006BAK02A05);农业科技跨越计划项目(2005-4);教育部新世纪优秀人才支持计划项目(2005);陕西省重大科技专项项目(2006KZ09-G1)

摘　　要：【目的】选用辅酶Q10产生菌深红红螺菌和根癌土壤杆菌为出发菌株,系统研究其原生质体制备和再生条件,为辅酶Q10的工业化生产提供技术支持。【方法】从菌龄、溶菌酶质量浓度、酶解时间、酶解温度、高渗溶液的选择等,对深红红螺菌和根癌土壤杆菌2株菌进行了原生质体制备和再生条件的研究。【结果】①根癌土壤杆菌原生质体制备和再生的适宜条件为:菌龄13.5 h,高渗溶液为0.8 mol/L NaCl,溶菌酶质量浓度为0.2 mg/mL,酶解时间75 min,酶解温度35℃;②深红红螺菌原生质体制备和再生的适宜条件为:菌龄45 h,高渗溶液为0.5 mol/L蔗糖+0.02 mol/L的MgCl2.6H2O,溶菌酶质量浓度为3 mg/L,酶解时间60 min,酶解温度37℃。【结论】得到了根癌土壤杆菌AT-N10和深红红螺菌Rh1.5005适宜的原生质体制备和再生条件。[Objective] Using Agrobacterium tumefaciens and Rhodospirillurn rubrum as original strains,the research studied the proper protoplast preparation and regeneration conditions for providing CoQ10 scale-up process with technical support. [Method] The protoplast preparation and regeneration conditions of Agrobacteriurn turnefaciens and Rhodospirillum rubrum were studied from strain age, enzyme concentration,enzymatic time, enzymatic temperature and the certain of hypertonic solution. [Resultl （DThe proper conditions of protoplast preparation and regeneration for Agrobacterium tumefaciens ; 13.5 h aged, hypertonic solution 0.8 mol/L NaCl, enzymatic time 75 min, enzymatic temperature 35 ℃, enzymatic concentration 0.2 mg/mL; （2）The proper conditions of protoplast preparation and regeneration for Rho- dospirillum rubrum .45 h aged,hypertonic solution 0.5 mol/L sucrose and 0.02 mol/L MgC12 · 6H2O, en- zymatic time 60 min, enzymatic temperature 37 ℃, enzymatic concentration 3 mg/L. [Conclusion] The proper conditions were obtained for protoplast preparation and regeneration of Agrobacterium tumefaciens and Rhodospirillum rubrum. This would he the basis of protoplast fusion of these two strains.

关 键 词：CoQ10/辅酶Q10 根癌土壤杆菌 深红红螺菌 原生质体制备 再生条件 

分 类 号：Q813.2[生物学—生物工程]