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[1]于平,徐超超,朱鹏志.产碱性果胶酶的枯草芽孢杆菌ZGL14原生质体制备与再生条件[J].应用与环境生物学报,2018,24(02):357-361.[doi:10.19675/j.cnki.1006-687x.2017.05040]
 YU Ping XU Chaochao,ZHU Pengzhi.Protoplast preparation and regeneration conditions of Bacillus subtilis ZGL14 producing alkaline pectinase[J].Chinese Journal of Applied & Environmental Biology,2018,24(02):357-361.[doi:10.19675/j.cnki.1006-687x.2017.05040]
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产碱性果胶酶的枯草芽孢杆菌ZGL14原生质体制备与再生条件()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
24卷
期数:
2018年02期
页码:
357-361
栏目:
研究论文
出版日期:
2018-04-25

文章信息/Info

Title:
Protoplast preparation and regeneration conditions of Bacillus subtilis ZGL14 producing alkaline pectinase
作者:
于平徐超超朱鹏志
浙江工商大学食品与生物工程学院 杭州 310035
Author(s):
YU Ping XU Chaochao ZHU Pengzhi
College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310035, China
关键词:
枯草芽孢杆菌原生质体制备原生质体再生形成率再生率渗透压稳定剂
Keywords:
Bacillus subtilis protoplast preparation protoplast regeneration formation rate regeneration rate osmotic pressure stabilizer
分类号:
Q813.2
DOI:
10.19675/j.cnki.1006-687x.2017.05040
摘要:
原生质体融合技术是一种重要的微生物育种方法. 以能产碱性果胶酶的枯草芽孢杆菌ZGL14为出发菌株,探讨影响其原生质体制备和再生的主要因素. 通过研究菌龄、溶菌酶浓度、酶解时间、酶解温度、渗透压稳定剂的pH值、再生培养基类型和培养方式等对枯草芽孢杆菌ZGL14原生质体制备与再生的影响,确定其最优化条件. 结果显示:枯草芽孢杆菌ZGL14原生质体制备的最优条件为菌龄9 h,溶菌酶浓度0.2 mg/mL,酶解时间10 min,酶解温度32 ℃,渗透压稳定剂的pH值 7.0. 在此条件下,枯草芽孢杆菌ZGL14原生质体的形成率为98%,再生率达30%. 枯草芽孢杆菌ZGL14原生质体再生培养基以琥珀酸钠作为渗透压稳定剂并结合Ca2+再生效果较好,再生方式以单层培养较好. 本研究获得了枯草芽孢杆菌ZGL14原生质体制备和再生的最佳条件,可为实现其融合育种提供技术支持. (图7 表1 参14)
Abstract:
The protoplast fusion technique is one of the important microbial breeding methods. The main factors that affect protoplast preparation and regeneration were investigated using Bacillus subtilis ZGL14 as the starter strain. The optimal conditions for protoplast preparation and regeneration were determined in this study by optimizing the cell age, lysozyme concentration, enzymolysis time, enzymolysis temperature, pH values of the osmotic pressure stabilizer, regeneration medium types, and culture type. The results indicated that the optimal condition for protoplast preparation of B. subtilis ZGL14 was as follows: 9 h of cell age, 0.2 mg/mL of lysozyme concentration, 10 min of enzymolysis time, 32 °C of enzymolysis temperature, and 7.0 of osmotic pressure stabilizer pH value. Under this condition, the protoplast formation and regeneration rates were > 98% and > 30%, respectively. A good protoplast regeneration effect can be achieved using the regeneration medium with sodium succinate as osmotic pressure stabilizer in combination with Ca2+. The single-layer culture was better than double-layer for regeneration. The optimal condition for protoplast preparation and regeneration of B. subtilis ZGL14 was determined in this study, which provides technical support for protoplast fusion breeding of B. subtilis.

参考文献/References:

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更新日期/Last Update: 2018-04-25