| 多杀菌素生产菌原生质体的制备条件及其再生菌株生理特性 |
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| 引用本文： | 罗玉双,丁学知,夏立秋,王海龙,黄璠,唐滢.多杀菌素生产菌原生质体的制备条件及其再生菌株生理特性[J].生物工程学报,2009,25(3):360-367. |
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| 作者姓名： | 罗玉双 丁学知 夏立秋 王海龙 黄璠 唐滢 |
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| 作者单位： | 湖南师范大学生命科学学院,微生物分子生物学湖南省重点实验室,长沙,410081 |
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| 基金项目： | 国家“863计划”(Nos.2006AA02Z187,2006AA10A212);;教育部博士点基金(No.20060542006);;国家自然科学基金项目(Nos.30870064,30670052)资助~~ |
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| 摘 要： | 为了改良多杀菌素生产菌种,提高多杀菌素产量,研究了甘氨酸添加浓度、溶菌酶作用时间、温度和浓度对多杀菌素生产菌刺糖多胞菌Saccharopolyspora spinosaSP06081菌株原生质体制备和再生的影响,并考察了不同再生培养基和渗透压稳定剂对其再生的影响,确定了该菌株原生质体制备和再生的最佳条件。同时,对原生质体再生菌株的形态与多杀菌素产量变化进行了比较研究。结果表明:菌体在添加0.2%的甘氨酸的TSB培养基中培养48h收集,0.1mg/mL溶菌酶,28oC作用20min制备原生质体,将原生质体涂布于以蔗糖为渗透压稳定剂的R2YE培养基中,原生质体再生数目最多,达108个/mL以上;原生质体再生菌株在形态和抗生素产量上产生分化,29.3%的再生菌株形态上保持与亲本菌株一致,具有菌丝松散,断裂分枝多的特点,其中53.2%的再生菌株多杀菌素产量变异向正方向移动,最高产量达到582.0mg/L,比亲本菌株提高85.6%。原生质体再生菌株的形态分化与多杀菌素产量具有重要相关性。
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| 关 键 词： | 刺糖多胞菌 多杀菌素 原生质体 再生菌株 形态分化 |
| 收稿时间： | 2008/9/26 0:00:00 |
Conditions for protoplast preparation of spinosyn-producing strain and the physiological properties of protoplast-regenerated strains |
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| Yushuang Luo,Xuezhi Ding,Liqiu Xi,Hailong Wang,Fan Huang and Ying Tang.Conditions for protoplast preparation of spinosyn-producing strain and the physiological properties of protoplast-regenerated strains[J].Chinese Journal of Biotechnology,2009,25(3):360-367. |
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| Authors: | Yushuang Luo Xuezhi Ding Liqiu Xi Hailong Wang Fan Huang and Ying Tang |
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| Institution: | Key Laboratory of Microbial Molecular Biology of Hunan Province, College of Life Science, Hunan Normal University, Changsha 410081, China;Key Laboratory of Microbial Molecular Biology of Hunan Province, College of Life Science, Hunan Normal University, Changsha 410081, China;Key Laboratory of Microbial Molecular Biology of Hunan Province, College of Life Science, Hunan Normal University, Changsha 410081, China;Key Laboratory of Microbial Molecular Biology of Hunan Province, College of Life Science, Hunan Normal University, Changsha 410081, China;Key Laboratory of Microbial Molecular Biology of Hunan Province, College of Life Science, Hunan Normal University, Changsha 410081, China;Key Laboratory of Microbial Molecular Biology of Hunan Province, College of Life Science, Hunan Normal University, Changsha 410081, China |
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| Abstract: | To improve spinosyn-producing strain and enhance spinosyns yield, we studied the effects of glycin concentration and the operational time, temperature and lysozyme concentration on protoplast preparation of Saccharopolyspora spinosa SP06081. We also studied different regeneration media and osmotic stabilizing agents. In addition, we compared the change of morphology and spinosyns yield of the regenerated strains. The results showed that the Saccharopolyspora spinosa SP06081 protoplast yield was the highest ... |
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| Keywords: | Saccharopolyspora spinosa spinosyns protoplast protoplast-regenerated strains morphological differentiation |
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