Growth of E-coli BL21 in minimal media with different gluconeogenic carbon sources and salt contents
Growth of E-coli BL21 in minimal media with different gluconeogenic carbon sources and salt contents
复制标题
DOI：
10.1007/s00253-006-0554-8
复制
发表时间：
2007-01-01
影响因子：
5
通讯作者：
Gunasekera, Thusitha S.
中科院分区：
文献类型：
--
作者：
Paliy, Oleg;Gunasekera, Thusitha S.
大肠杆菌菌株BL 21通常用作蛋白质表达和纯化的宿主菌株。对于结构分析，蛋白质通常通过生长E.大肠杆菌在含有适当同位素的培养基中培养。当需要大量的全氘代蛋白质时，E.大肠杆菌通常在以氘代琥珀酸盐或乙酸盐作为碳源的基本培养基中生长，因为这些较便宜。尽管BL21被广泛使用，但我们没有发现关于不同基本培养基和碳源对BL21生长的影响的数据。在这项研究中，我们评估了E。coli BL21基本培养基中的发酵条件。虽然BL21在甘油和丙酮酸上生长良好，但它在琥珀酸（20 h）、乙酸（10 h）和富马酸（20 h）上具有较长的滞后期，这归因于E. coli细胞。野生型菌株NCM3722（K12）在所有底物上生长良好。我们还研究了E. coliBL21在基本培养基中的盐组成不同，但碳源不同。常用的M9培养基不支持E. coli BL21基本培养基中进行发酵。在M9培养基中添加硫酸亚铁（不添加硫酸亚铁）可提高E.大肠杆菌培养物，并显着增加其细胞密度在稳定期。
Escherichia coli strain BL21 is commonly used as a host strain for protein expression and purification. For structural analysis, proteins are frequently isotopically labeled with deuterium (H-2), C-13, or N-15 by growing E. coli cultures in a medium containing the appropriate isotope. When large quantities of fully deuterated proteins are required, E. coli is often grown in minimal media with deuterated succinate or acetate as the carbon source because these are less expensive. Despite the widespread use of BL21, we found no data on the effect of different minimal media and carbon sources on BL21 growth. In this study, we assessed the growth behavior of E. coli BL21 in minimal media with different gluconeogenic carbon sources. Though BL21 grew reasonably well on glycerol and pyruvate, it had a prolonged lag-phase on succinate (20 h), acetate (10 h), and fumarate (20 h), attributed to the physiological adaptation of E. coli cells. Wild-type strain NCM3722 (K12) grew well on all the substrates. We also examined the growth of E. coli BL21 in minimal media that differed in their salt composition but not in their source of carbon. The commonly used M9 medium did not support the optimum growth of E. coli BL21 in minimal medium. The addition of ferrous sulphate to M9 medium (otherwise lacking it) increased the growth rate of E. coli cultures and significantly increased their cell density in the stationary phase.