Smart sustainable bottle (SSB) system for E. coli based recombinant protein production.
Smart sustainable bottle (SSB) system for E. coli based recombinant protein production.
复制标题
DOI：
10.1186/s12934-014-0153-9
复制
发表时间：
2014-11-05
影响因子：
6.4
通讯作者：
Rinas U
中科院分区：
文献类型：
--
作者：
Li Z;Carstensen B;Rinas U
对蛋白质感兴趣但对生产过程本身不感兴趣的实验室通常需要重组蛋白质。因此，易于操作的技术设备和直接的蛋白质生产程序对这些实验室来说是非常有利的。销售一次性培养袋和生物反应器的公司正在努力满足至少部分这些需求。然而，一次性系统可能会产生重大成本，当需要“良好生产规范”时，这可能是可以接受的，但对于大多数面临资金紧张的实验室来说是不可接受的。介绍了一种简单的自制“智能可持续瓶”（SSB）系统的组装和应用，用于大肠杆菌蛋白生产。 SSB系统的核心是一个2L玻璃瓶，在恒温、气流和搅拌速度下运行，无需测量和控制pH和溶解氧。氧转移能力处于以中等通气速率运行的传统生物反应器的范围内，并且远远超过传统摇瓶和一次性生物反应器中的氧转移能力。 SSB 系统用于使用基于 T7 的表达系统和确定的自诱导培养基生产各种重组蛋白。蛋白质的量和溶解度方面的生产性能具有稳健和细腻的特性，与最先进的搅拌罐商业生物反应器一样好。 SSB 系统代表了一种低成本的蛋白质生产装置，适用于简单、有效和可重复的重组蛋白质生产。本文的在线版本 (doi:10.1186/s12934-014-0153-9) 包含补充材料，可供授权用户使用。
Recombinant proteins are usually required in laboratories interested in the protein but not in the production process itself. Thus, technical equipment which is easy to handle and straight forward protein production procedures are of great benefit to those laboratories. Companies selling single use cultivation bags and bioreactors are trying to satisfy at least part of these needs. However, single-use systems can contribute to major costs which might be acceptable when “good manufacturing practices” are required but not acceptable for most laboratories facing tight funding. The assembly and application of a simple self-made “smart sustainable bottle” (SSB) system for E. coli based protein production is presented. The core of the SSB system is a 2-L glass bottle which is operated at constant temperature, air flow, and stirrer speed without measurement and control of pH and dissolved oxygen. Oxygen transfer capacities are in the range as in conventional bioreactors operated at intermediate aeration rates and by far exceed those found in conventional shaking flasks and disposable bioreactors. The SSB system was applied for the production of various recombinant proteins using T7-based expression systems and a defined autoinduction medium. The production performance regarding amount and solubility of proteins with robust and delicate properties was as good as in state-of-the-art stirred tank commercial bioreactors. The SSB system represents a low cost protein production device applicable for easy, effective, and reproducible recombinant protein production. The online version of this article (doi:10.1186/s12934-014-0153-9) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
--
作者：
Cooper, CM;Fernstrom, GA;Miller, SA
通讯作者：
Miller, SA
影响因子：
2.9
作者：
Hoffmann, F;Rinas, U
通讯作者：
Rinas, U
影响因子：
1.6
作者：
Tomala, Magda;Lavrentieva, Antonina;Scheper, Thomas
通讯作者：
Scheper, Thomas
影响因子：
2.9
作者：
Candiano, G;Bruschi, M;Righetti, PG
通讯作者：
Righetti, PG
影响因子：
5.6
作者：
Kloeckner, Wolf;Gacem, Riad;Buechs, Jochen
通讯作者：
Buechs, Jochen