| 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、酶解时问、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/LNaCl,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MgC12·6H2O,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. |
| 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 |
| DOI： |
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| 基金项目:国家科技支撑计划，农业科技跨越计划，教育部新世纪优秀人才支持计划，陕西省重大科技专项计划 |
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| Study on preparation and regeneration of protoplasts from CoQ10-yield strains |
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| Abstract: |
| 【Objective】 Using Agrobacterium tumefacien and Rhodospirillum 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 Agrobacterium tumefaciens and Rhodospirillum rubrum were studied from strain age,enzyme concentration,enzymatic time,enzymatic temperature and the certain of hypertonic solution.【Result】 ①The 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;②The proper conditions of protoplast preparation and regeneration for Rhodospirillum rubrum:45 h aged,hypertonic solution 0.5 mol/L sucrose and 0.02 mol/L MgCl2·6H2O, enzymatic 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 be the basis of protoplast fusion of these two strains. |
| Key words: CoQ10/coenzyme Q10 Agrobacterium tumefaciens Rhodospirillum rubrum protoplast preparation regeneration of protoplast |
