Enhanced synthesis of alginate oligosaccharides in Pseudomonas mendocina NK-01 by overexpressing MreB
Enhanced synthesis of alginate oligosaccharides in Pseudomonas mendocina NK-01 by overexpressing MreB
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通过过表达 MreB 增强门多萨假单胞菌 NK-01 中藻酸盐寡糖的合成
DOI：
10.1007/s13205-019-1873-7
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发表时间：
2019-08
期刊：
影响因子：
2.8
通讯作者：
Yang Chao
中科院分区：
文献类型：
--
作者：
Fan Xu;Gong Ting;Wu Yunbo;Zhao Fengjie;Qiao Mingqiang;Wang Shufang;Yang Chao
本研究旨在研究细胞骨架蛋白MreB对Mendocina假单胞菌NK-01细菌细胞形态及藻酸低聚糖(AO)和聚羟基烷酸(PHA)合成的影响。为过表达mreB基因，构建了编码mreB-GFP融合蛋白的表达载体。扫描电子显微镜显示，表达MreB的细胞比野生的细胞更长，这与MreB与肽聚糖合成的关系是一致的。表达MreB-GFP融合蛋白的细胞在荧光显微镜下发出绿色荧光，表明MreB在NK-01菌株中得到了功能性表达。激光共聚焦显微镜下可见MreB定位于细胞膜周围。此外，重组菌株可合成0.961 g/L的AO，是野生型菌株的5.86倍。通过培养基优化试验，最终确定以20g/L葡萄糖为最佳糖原添加量，以获得较高的AO产量和较高的底物转化率。结果表明，MreB基因的过表达影响了细胞形态、AO聚合酶活性和AO分泌效率。但是，重组菌株的PHA合成略有减少。结果表明，这种细胞骨架蛋白的过表达影响了特定胞内和胞外产物的产量。
This study aimed to investigate the effects of cytoskeleton protein MreB on bacterial cell morphology and the synthesis of alginate oligosaccharides (AO) and polyhydroxyalkanoate (PHA) by Pseudomonas mendocina NK-01. To overexpress the mreB gene, an expression vector encoding MreB-GFP fusion protein was constructed. The scanning electron microscope (SEM) showed that cells expressing MreB were longer than the wild ones, which agrees with MreB’s relationship with the synthesis of peptidoglycan. Cells expressing the MreB-GFP fusion protein emitted green fluorescence under a fluorescence microscope, suggesting that MreB was functionally expressed in strain NK-01. Under a confocal laser scanning microscope, MreB was observed as located around the cell membrane. Furthermore, the recombinant strain could synthesize 0.961 g/L AO, which was 5.86-fold higher than wild-type strain. Through the medium optimization test, we finally selected the addition of 20 g/L glucose as the optimal glycogen addition for AO fermentation based on a high AO yield and high substrate transformation efficiency. The results indicated that overexpression of MreB affected the cell morphology, the activity of AO polymerase, and the efficiency of AO secretion. However, the synthesis of PHA for recombinant strain was slightly reduced. The results suggested that the overexpression of this cytoskeleton protein affected the yield of specific intracellular and extracellular products.
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影响因子：
11.4
作者：
Kruse, T;Moller-Jensen, J;Gerdes, K
通讯作者：
Gerdes, K
影响因子：
11.4
作者：
Shiomi, Daisuke;Sakai, Masako;Niki, Hironori
通讯作者：
Niki, Hironori
影响因子：
11.1
作者：
Nan B;Zusman DR
通讯作者：
Zusman DR
DOI：
10.1073/pnas.0402638101
发表时间：
2004-06-08
影响因子：
11.1
作者：
Gitai, Z;Dye, N;Shapiro, L
通讯作者：
Shapiro, L
影响因子：
3.9
作者：
OTTERLEI, M;OSTGAARD, K;ESPEVIK, T
通讯作者：
ESPEVIK, T