Development of an optimized medium, strain and high-throughput culturing methods for Methylobacterium extorquens.
Development of an optimized medium, strain and high-throughput culturing methods for Methylobacterium extorquens.
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开发针对甲甲基甲虫的优化培养基，菌株和高通量培养方法。
DOI：
10.1371/journal.pone.0062957
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Marx CJ
中科院分区：
文献类型：
--
作者：
Delaney NF;Kaczmarek ME;Ward LM;Swanson PK;Lee MC;Marx CJ
扭脱甲基杆菌菌株是研究得最好的甲基营养模式系统，其单碳化合物代谢的研究已经超过50年。在这里，我们开发了一个新的系统，用于高通量分批培养M。通过共同优化生物体、生长培养基和培养系统的性质，在微量滴定板中培养异育螺旋体。去除M.为了防止生物膜形成，我们发现，目前可用的实验室自动化设备，集成和开源软件管理，使得指数增长率的可靠估计成为可能。利用该系统，我们开发了一种优化的M.使用响应面方法的解旋。我们发现使用EDTA作为金属螯合剂的培养基抑制生长并导致不一致的培养条件。相比之下，我们用PIPES缓冲液和柠檬酸盐螯合的金属开发的新培养基允许快速和更一致的生长速率。这种新的甲基杆菌PIPES（“MP”）培养基对于其组分成分的大偏差也是稳健的，这避免了来自使用在不同时间制备的培养基的实验的批次效应。MP培养基允许比用于M的其他培养基更快和更一致的生长。勒索。
Methylobacterium extorquens strains are the best-studied methylotrophic model system, and their metabolism of single carbon compounds has been studied for over 50 years. Here we develop a new system for high-throughput batch culture of M. extorquens in microtiter plates by jointly optimizing the properties of the organism, the growth media and the culturing system. After removing cellulose synthase genes in M. extorquens strains AM1 and PA1 to prevent biofilm formation, we found that currently available lab automation equipment, integrated and managed by open source software, makes possible reliable estimates of the exponential growth rate. Using this system, we developed an optimized growth medium for M. extorquens using response surface methodologies. We found that media that used EDTA as a metal chelator inhibited growth and led to inconsistent culture conditions. In contrast, the new medium we developed with a PIPES buffer and metals chelated by citrate allowed for fast and more consistent growth rates. This new Methylobacterium PIPES (‘MP’) medium was also robust to large deviations in its component ingredients which avoided batch effects from experiments that used media prepared at different times. MP medium allows for faster and more consistent growth than other media used for M. extorquens.
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影响因子：
4.1
作者：
DALES, SL;ANTHONY, C
通讯作者：
ANTHONY, C
影响因子：
3.7
作者：
Kiefer, Patrick;Buchhaupt, Markus;Vorholt, Julia A.
通讯作者：
Vorholt, Julia A.
影响因子：
4.1
作者：
LARGE, PJ;QUAYLE, JR;PEEL, D
通讯作者：
PEEL, D
影响因子：
64.5
作者：
Bollenbach T;Quan S;Chait R;Kishony R
通讯作者：
Kishony R
影响因子：
4.5
作者：
Lee MC;Marx CJ
通讯作者：
Marx CJ