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在一株工业菌株中,L-阿拉伯糖通过诱导阿拉伯糖特异性Gal2p转运蛋白来触发自身的摄取。

l-Arabinose triggers its own uptake via induction of the arabinose-specific Gal2p transporter in an industrial strain.

作者信息

Oehling Verena, Klaassen Paul, Frick Oliver, Dusny Christian, Schmid Andreas

机构信息

1Laboratory of Chemical Biotechnology, Department of Biochemical & Chemical Engineering, TU Dortmund University, Dortmund, Germany.

DSM Biotechnology Center, Delft, The Netherlands.

出版信息

Biotechnol Biofuels. 2018 Aug 23;11:231. doi: 10.1186/s13068-018-1231-8. eCollection 2018.

Abstract

Bioethanol production processes with using lignocellulosic biomass as feedstock are challenged by the simultaneous utilization of pentose and hexose sugars from biomass hydrolysates. The pentose uptake into the cell represents a crucial role for the efficiency of the process. The focus of the here presented study was to understand the uptake and conversion of the pentose l-arabinose in and reveal its regulation by d-glucose and d-galactose. Gal2p-the most prominent transporter enabling l-arabinose uptake in wild-type strains-has an affinity for the transport of l-arabinose, d-glucose, and d-galactose. d-Galactose was reported for being mandatory for inducing expression. expression is also known to be regulated by d-glucose-mediated carbon catabolite repression, as well as catabolite inactivation. The results of the present study demonstrate that l-arabinose can be used as sole carbon and energy source by the recombinant industrial strain DS61180. RT-qPCR and RNA-Seq experiments confirmed that l-arabinose can trigger its own uptake via the induction of expression. Expression levels of during growth on l-arabinose reached up to 21% of those obtained with d-galactose as sole carbon and energy source. l-Arabinose-induced expression was also subject to catabolite repression by d-glucose. Kinetic investigations of substrate uptake, biomass, and product formation during growth on a mixture of d-glucose/l-arabinose revealed impairment of growth and ethanol production from l-arabinose upon d-glucose depletion. The presence of d-glucose is thus preventing the fermentation of l-arabinose in DS61180. Comparative transcriptome studies including the wild-type and a precursor strain delivered hints for an increased demand in ATP production and cofactor regeneration during growth of DS61180 on l-arabinose. Our results thus emphasize that cofactor and energy metabolism demand attention if the combined conversion of hexose and pentose sugars is intended, for example in biorefineries using lignocellulosics.

摘要

以木质纤维素生物质为原料的生物乙醇生产工艺面临着同时利用生物质水解液中的戊糖和己糖的挑战。戊糖进入细胞对该工艺的效率起着关键作用。本研究的重点是了解戊糖L-阿拉伯糖的摄取和转化,并揭示其受D-葡萄糖和D-半乳糖的调控。Gal2p是野生型菌株中使L-阿拉伯糖摄取的最主要转运蛋白,它对L-阿拉伯糖、D-葡萄糖和D-半乳糖的转运具有亲和力。据报道,D-半乳糖是诱导表达所必需的。已知表达也受D-葡萄糖介导的碳分解代谢物阻遏以及分解代谢物失活的调控。本研究结果表明,重组工业菌株DS61180可以将L-阿拉伯糖用作唯一的碳源和能源。RT-qPCR和RNA-Seq实验证实,L-阿拉伯糖可通过诱导表达来触发自身摄取。在以L-阿拉伯糖为碳源和能源生长期间,的表达水平达到以D-半乳糖为唯一碳源和能源时所获表达水平的21%。L-阿拉伯糖诱导的表达也受到D-葡萄糖的分解代谢物阻遏。对在D-葡萄糖/L-阿拉伯糖混合物上生长期间底物摄取、生物质和产物形成的动力学研究表明,在D-葡萄糖耗尽后,L-阿拉伯糖的生长和乙醇生产受到损害。因此,D-葡萄糖的存在阻止了DS61180中L-阿拉伯糖的发酵。包括野生型和前体菌株的比较转录组研究为DS61180在L-阿拉伯糖上生长期间ATP产生和辅因子再生需求增加提供了线索。因此,我们的结果强调,如果要实现己糖和戊糖的联合转化,例如在使用木质纤维素的生物精炼厂中,辅因子和能量代谢需要引起关注。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23b6/6106821/45d9fd79f891/13068_2018_1231_Fig1_HTML.jpg

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