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在基因修饰的大肠杆菌中重组合成细菌纤维素。

Recombinant biosynthesis of bacterial cellulose in genetically modified Escherichia coli.

机构信息

Biological Systems Engineering Laboratory (BSEL), Department of Chemical Engineering, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK.

Department of Bioengineering, Marmara University, Göztepe Campus, Istanbul, Turkey.

出版信息

Bioprocess Biosyst Eng. 2018 Feb;41(2):265-279. doi: 10.1007/s00449-017-1864-1. Epub 2017 Nov 24.

Abstract

Bacterial cellulose (BC) exhibits unique properties such as high purity compared to plant-based cellulose; however, commercial production of BC has remained a challenge, primarily due to the strain properties of cellulose-producing bacteria. Herein, we developed a functional and stable BC production system in genetically modified (GM) Escherichia coli by recombinant expression of both the BC synthase operon (bcsABCD) and the upstream operon (cmcax, ccp Ax). BC production was achieved in GM HMS174 (DE3) and in GM C41 (DE3) by optimization of the culture temperature (22 °C, 30 °C, and 37 °C) and IPTG concentration. BC biosynthesis was detected much earlier in GM C41 (DE3) cultures (3 h after IPTG induction) than those of Gluconacetobacter hansenii. GM HMS174 (DE3) produced dense fibres having a length of approximately 1000-3000 μm and a diameter of 10-20 μm, which were remarkably larger than the fibres of BC typically produced by G. hansenii.

摘要

细菌纤维素 (BC) 相较于植物纤维素具有高纯度等独特性质,但商业生产 BC 仍然具有挑战性,主要是由于产纤维素细菌的菌株特性所致。在此,我们通过重组表达 BC 合酶操纵子 (bcsABCD) 和上游操纵子 (cmcax、ccpAx),在遗传修饰 (GM) 的大肠杆菌中开发了一个功能稳定的 BC 生产系统。通过优化培养温度(22°C、30°C 和 37°C)和 IPTG 浓度,在 GM HMS174(DE3)和 GM C41(DE3)中实现了 BC 的生产。在 GM C41(DE3)培养物中(IPTG 诱导后 3 小时)更早检测到 BC 生物合成,而在 Gluconacetobacter hansenii 中则较晚。GM HMS174(DE3)产生的致密纤维长度约为 1000-3000μm,直径为 10-20μm,明显大于通常由 G. hansenii 产生的 BC 纤维。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da8c/5773641/c0617c74d5c5/449_2017_1864_Fig1_HTML.jpg

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