Succinic acid production from sucrose and molasses by metabolically engineered E. coli using a cell surface display system
Succinic acid production from sucrose and molasses by metabolically engineered E. coli using a cell surface display system
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使用细胞表面显示系统通过代谢工程大肠杆菌从蔗糖和糖蜜生产琥珀酸
DOI：
10.1016/j.bej.2014.08.014
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发表时间：
2014-10-15
影响因子：
3.9
通讯作者：
Ouyang, Pingkai
中科院分区：
文献类型：
--
作者：
Ma, Jiangfeng;Li, Feng;Ouyang, Pingkai
为了实现蔗糖代谢能力，已将不同的蔗糖利用操纵子引入不能利用蔗糖的大肠杆菌中。然而，这些工程菌株仍然存在低生长率和低蔗糖摄取率的问题。在本研究中，细胞表面展示系统被应用于工程大肠杆菌AFP111中，用于直接从蔗糖和糖蜜生产琥珀酸。来自大肠杆菌 W 的转化酶 (CscA) 通过与 OmpC 锚定基序融合成功锚定到外膜，并且展示的 CscA 显示出高细胞外活性。与蔗糖通透酶系统相比，细胞表面展示系统在蔗糖代谢过程中消耗的ATP更少。当AFP111/pTrcC-cscA消耗较少的ATP时，来自蔗糖的琥珀酸生产率比具有蔗糖渗透酶系统的AFP111/pCR2.1-cscBKA高23%。结果，在双相发酵过程中，AFP111/pTrcC-cscA 在 3 L 发酵罐中 34 h 时分别从蔗糖和甘蔗糖蜜中产生了 41 g L-1 和 36.3 g L-1 琥珀酸。此外，在3L发酵罐中双相发酵结束时，回收的AFP111/pTrcC-cscA细胞积累了79gL-1琥珀酸，总产量为1.19molmol(-1)己糖。 (C) 2014 Elsevier B.V. 保留所有权利。
To achieve sucrose-metabolizing capability, different sucrose utilization operons have been introduced into E. con that cannot utilize sucrose. However, these engineered strains still suffer from low growth rates and low sucrose uptake rates. In this study, cell surface display system was adopted in engineered E. coli AFP111 for succinic acid production from sucrose and molasses directly. Invertase (CscA) from E. coli W was successfully anchored to outer membrane by fusion with OmpC anchoring motif, and the displayed CscA showed high extracellular activity. Compared with the sucrose permease system, the cell surface display system consumed less ATP during sucrose metabolism. When less ATP was consumed by AFP111/pTrcC-cscA, the succinic acid productivity from sucrose was 23% higher than that by AFP111/pCR2.1-cscBKA that having the sucrose permease system. As a result, 41 g L-1 and 36.3 g L-1 succinic acid were produced by AFP111/pTrcC-cscA from sucrose and sugarcane molasses respectively at 34 h in 3-L fermentor during dual-phase fermentation. In addition, 79 g L-1 succinic acid was accumulated with recovered AFP111/pTrcC-cscA cells at the end of dual-phase fermentation in 3-L fermentor, and the overall yield was 1.19 mol mol(-1) hexose. (C) 2014 Elsevier B.V. All rights reserved.