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嗜蜚希瓦氏菌(p424IbPSO)的静息细胞产异丁醇:生长培养条件的影响

Resting cells isobutanol production by Shimwellia blattae (p424IbPSO): Influence of growth culture conditions.

作者信息

Acedos Miguel G, Santos Victoria E, Garcia-Ochoa Felix

机构信息

Dept. of Chemical and Materials Engineering, Universidad Complutense, Madrid, 28040, Spain.

出版信息

Biotechnol Prog. 2018 Sep;34(5):1073-1080. doi: 10.1002/btpr.2705. Epub 2018 Oct 3.

Abstract

Isobutanol is a promising gasoline additive and could even be a potential substitute used directly as combustible. In this work, the production of isobutanol from glucose by Shimwellia blattae (p424IbPSO) in resting cell cultures is studied. This production has two stages, involving a resting cell phase that has not been studied before. The cell growth was carried out under different operating conditions: temperature and medium composition (YE, ammonium, and IPTG concentrations), looking for the highest isobutanol production. Moreover, the cells were collected at three different growth times checking their isobutanol production capacity. The best operating conditions have been determined as: 30°C of temperature, a medium containing 1.5 g L YE and 1.4 g L of ammonium as nitrogen sources, adding 0.5 mM IPTG as inducer. The cells collected at early growth times are significantly more active. The use of S. blattae (p424IbPSO) in resting cells is a good strategy for the production of isobutanol from glucose yielding better results than in batch growth cultures, a yield of 60% attainment of theoretical maximum yield is obtained under optimal conditions. In addition, it has been demonstrated that if the cells are cultured at higher temperatures and with high IPTG concentrations, inclusion bodies are formed in the cytoplasm inhibiting the isobutanol production in the resting cell stage.

摘要

异丁醇是一种很有前景的汽油添加剂,甚至可能成为直接用作燃料的潜在替代品。在这项工作中,研究了静止细胞培养中谢氏蜚蠊(p424IbPSO)由葡萄糖生产异丁醇的过程。该生产过程有两个阶段,其中包括一个此前未被研究过的静止细胞阶段。细胞生长在不同的操作条件下进行:温度和培养基组成(酵母提取物、铵和异丙基硫代半乳糖苷浓度),以寻找最高的异丁醇产量。此外,在三个不同的生长时间收集细胞,检测它们生产异丁醇的能力。已确定最佳操作条件为:温度30°C,含有1.5 g/L酵母提取物和1.4 g/L铵作为氮源的培养基,添加0.5 mM异丙基硫代半乳糖苷作为诱导剂。在生长早期收集的细胞活性明显更高。在静止细胞中使用谢氏蜚蠊(p424IbPSO)是由葡萄糖生产异丁醇的一种良好策略,比分批生长培养产生更好的结果,在最佳条件下可获得60%的理论最大产量。此外,已经证明,如果细胞在较高温度和高异丙基硫代半乳糖苷浓度下培养,细胞质中会形成包涵体,抑制静止细胞阶段的异丁醇生产。

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