Optimization of the Lactococcus lactis nisin-controlled gene expression system NICE for industrial applications.
Optimization of the Lactococcus lactis nisin-controlled gene expression system NICE for industrial applications.
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DOI：
10.1186/1475-2859-4-16
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发表时间：
2005-05-30
影响因子：
6.4
通讯作者：
Smid EJ
中科院分区：
文献类型：
--
作者：
Mierau I;Olieman K;Mond J;Smid EJ
乳酸乳球菌的乳链菌肽控制的基因表达系统NICE是革兰氏阳性菌中应用最广泛的表达系统之一。尽管其广泛使用，但尚未优化培养条件和乳链菌肽诱导以获得最大产量。作为诱导产生溶葡萄球菌素的模型系统，溶葡萄球菌素是一种由模拟葡萄球菌生物变种产生的抗菌蛋白（主要针对金黄色葡萄球菌）。使用了溶葡萄球菌。需要优化三个主要领域以获得最大产量：细胞密度、乳链菌肽控制的诱导和蛋白质生产以及目标蛋白质的特定参数。在一系列 pH 控制的发酵中，对以下参数进行了优化：培养物的 pH 值、NaOH 或 NH4OH 作为中和剂的使用、锌和磷酸盐的添加、发酵温度、诱导时间点（培养物的细胞密度）、用于诱导的乳链菌肽添加量以及三种基本培养基成分（即酵母提取物、蛋白胨和乳糖）的量。跟踪每种培养物的生长和溶葡萄球菌素的产生。溶葡萄球菌素产量取决于所有变化的参数。在优化过程中，溶葡萄球菌素产量增加了三倍，从 100 mg/l 增加到 300 mg/l。乳酸乳球菌中 NICE 基因表达系统的蛋白质生产很大程度上取决于培养基组成、发酵参数和诱导时添加的乳链菌肽量。仔细优化关键参数可显着提高目标蛋白的产量。
The nisin-controlled gene expression system NICE of Lactococcus lactis is one of the most widely used expression systems in Gram-positive bacteria. Despite its widespread use, no optimization of the culture conditions and nisin induction has been carried out to obtain maximum yields. As a model system induced production of lysostaphin, an antibacterial protein (mainly against Staphylococcus aureus) produced by S. simulans biovar. Staphylolyticus, was used. Three main areas need optimization for maximum yields: cell density, nisin-controlled induction and protein production, and parameters specific for the target-protein. In a series of pH-controlled fermentations the following parameters were optimized: pH of the culture, use of NaOH or NH4OH as neutralizing agent, the addition of zinc and phosphate, the fermentation temperature, the time point of induction (cell density of the culture), the amount of nisin added for induction and the amount of three basic medium components, i.e. yeast extract, peptone and lactose. For each culture growth and lysostaphin production was followed. Lysostaphin production yields depended on all parameters that were varied. In the course of the optimization a three-fold increase in lysostaphin yield was achieved from 100 mg/l to 300 mg/l. Protein production with the NICE gene expression system in L. lactis strongly depends on the medium composition, the fermentation parameters and the amount of nisin added for induction. Careful optimization of key parameters lead to a significant increase in the yield of the target protein.
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影响因子：
4.4
作者：
Miyoshi, A;Poquet, I;Langella, P
通讯作者：
Langella, P
影响因子：
17.3
作者：
Hannig, G;Makrides, SC
通讯作者：
Makrides, SC
影响因子：
4.4
作者：
Cibik, R;Tailliez, P;Chapot-Chartier, MP
通讯作者：
Chapot-Chartier, MP
影响因子：
--
作者：
Bolotin, A;Ehrlich, SD;Sorokin, A
通讯作者：
Sorokin, A
影响因子：
4.4
作者：
Enouf, V;Langella, P;Corthier, G
通讯作者：
Corthier, G