产琥珀酸放线杆菌的原生质体制备与再生
Protoplast Preparation and Regeneration of Actinobacillus succinogenes
查看参考文献15篇
文摘
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基因组重组技术是一项重要的菌种改造技术,原生质体制备和再生是进行基因组重组的前提和基础. 目前少有关于产琥珀酸放线杆菌(Actinobacillus succinogenes)CGMCC2650原生质体研究的报道. 为了优化该菌的原生质体制备和再生条件,及利用基因组重组技术构建优良菌种提供参考,研究了甘氨酸预处理,菌龄,酶浓度,作用时间,温度对产琥珀酸放线杆菌原生质体制备和再生的影响,并考察了不同渗透压稳定剂对其再生的影响. 结果表明,菌体在添加了0.6mg/ml甘氨酸的TSB培养基中培养5h后收集,用SMM稀释到OD660=1.0,用0.025mg/ml溶菌酶在37℃下酶解45min制备原生质体,将原生质体涂布于含0.3mol/L蔗糖的再生培养基中,再生率最大,达到40.9%. 确定了产琥珀酸放线杆菌原生质体制备和再生的最佳条件,所用的原生质体制备的方法对琥珀酸的产生没有影响,这为进一步开展该菌的原生质体诱变及基因组重组等研究奠定了基础 |
其他语种文摘
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Genome shuffling was an important technology of strain improvement. The effects of glycin concentration, cell age, lysozyme concentration, the operational time and temperature wer conducted to optimize the conditions of protoplast formation and regeneration of Actinobacillus succinogenes CGMCC 2650.The effects of the different osmotic stabilizing agents on regeneration were also examed. The protoplast yield was the highest under these conditions: The bacteria was cultivated in Tryptic Soy Broth (TSB) medium containing 0.6 mg/ml glycine for 5 h, then collected and diluted with SMM until the optical density (660nm) reached 1.0.The diluent was treated by 0.025mg/ml lysozyme at 37℃ for 45 min and plated on the TSB medium with 0.3 mol/L sucrose as osmotic stabilizer. The regeneration rate was up to 40.9%. This provides the optimized conditions of protoplast preparation and regeneration for genome shuffling of A.succinogenes |
来源
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中国生物工程杂志
,2010,30(6):103-108 【核心库】
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关键词
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产琥珀酸放线杆菌
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琥珀酸
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原生质体
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基因组重组
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地址
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1.
中国矿业大学(北京)化学与环境工程学院, 北京, 100083
2.
中国科学院过程工程研究所, 生化工程国家重点实验室, 北京, 100190
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语种
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中文 |
文献类型
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研究性论文 |
ISSN
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1671-8135 |
学科
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微生物学 |
基金
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中国科学院知识创新工程项目
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文献收藏号
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CSCD:3963508
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参考文献 共
15
共1页
|
1.
Zeikus J G. Biotechnology of succinic acid production and markets for derived industrial products.
Appl Microbiol Biotechnol,1999,51:545-552
|
被引
77
次
|
|
|
|
2.
McKinlay J B. Determining Actinobacillus succinogenes metabolic pathways and fluxes by NMR and GC-MS analyses of ~(13)C -labeled metabolic product isotopomers.
Metabolic Engineering,2007,9(2):177-192
|
被引
22
次
|
|
|
|
3.
McKinlay J B. ~(13)C-metabolic flux analysis of Actinobacillus succinogenes fermentative metabolism at different NaHCO_3 and H_2 concentrations.
Metabolic Engineering,2008,10(1):55-68
|
被引
10
次
|
|
|
|
4.
Song H. Modeling of batch fermentation kinetics for succinic acid production by Mannheimia succiniciproducens.
Biochemical Engineering Journal,2008,40(1):107-115
|
被引
5
次
|
|
|
|
5.
Cotelesage J J. Crystal structure of Anaerobiospirillum succiniciproducens PEP carboxykinase reveals an important active site loop.
The International Journal of Biochemistry & Cell Biology,2005,37(9):1829-1837
|
被引
5
次
|
|
|
|
6.
Isar J. A statistical method for enhancing the production of succinic acid from Escherichia coli under anaerobic conditions.
Bioresource Technology,2006,97(13):1443-1448
|
被引
9
次
|
|
|
|
7.
Guettler M V.
Method for making succinic acid, bacterial variants for use in the process and methods for obtaining variants. US: 5573931,1996
|
被引
1
次
|
|
|
|
8.
郑璞. 环境因素对琥珀酸放线杆菌Actinobacillus succinogenes CGMCC 1593发酵生产丁二酸的影响.
生物工程学报,2008,24(6):1051-1055
|
被引
7
次
|
|
|
|
9.
Zhang Y X. Genome shuffling leads to rapid phenotypic improvement in bacteria.
Nature,2002,415:644-646
|
被引
87
次
|
|
|
|
10.
Patnaik R. Genome shuffling of Lactobacillus for improved acid tolerance.
Nat Biotechnol,2002,20:707-712
|
被引
59
次
|
|
|
|
11.
邢建民.
用于生产琥珀酸的放线杆菌和方法.中国, DIC091 10015,2009
|
被引
1
次
|
|
|
|
12.
Kai Zhao. Study on the preparation and regeneration of protoplast from taxol-producing fungus Nodulisporium sylviforme.
Nature and Science,2004,2(2):52-59
|
被引
1
次
|
|
|
|
13.
杨汝德.
现代工业微生物学教程,2006:272-285
|
被引
1
次
|
|
|
|
14.
刘永翔. 戴氏绿僵菌原生质体的形成及再生.
武汉大学学报(理工版),2004,5(S2):20-22
|
被引
2
次
|
|
|
|
15.
梁惠仪. 全基因组重排育种技术提高豆豉纤溶酶菌产酶量.
中国生物工程杂志,2007,27(10):39-43
|
被引
12
次
|
|
|
|
|