کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6494130 | 1418334 | 2018 | 55 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Combined engineering of disaccharide transport and phosphorolysis for enhanced ATP yield from sucrose fermentation in Saccharomyces cerevisiae
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کلمات کلیدی
ORFChemostatG418SPasev/v - V / Vw/v - W / VATP - آدنوزین تری فسفات یا ATPFacilitated diffusion - انتشار تسهیل شدهSucrose phosphorylase - فسفورلیس ساکارزphosphoglucomutase - فسفوگلوکوموتازYeast physiology - فیزیولوژی مخمرopen reading frame - قاب خواندن بازlysogeny broth - لیزوزومی گوگردSynthetic medium - محیط مصنوعیweight per volume - وزن در هر جلدbasepair - پایه
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Anaerobic industrial fermentation processes do not require aeration and intensive mixing and the accompanying cost savings are beneficial for production of chemicals and fuels. However, the free-energy conservation of fermentative pathways is often insufficient for the production and export of the desired compounds and/or for cellular growth and maintenance. To increase free-energy conservation during fermentation of the industrially relevant disaccharide sucrose by Saccharomyces cerevisiae, we first replaced the native yeast α-glucosidases by an intracellular sucrose phosphorylase from Leuconostoc mesenteroides (LmSPase). Subsequently, we replaced the native proton-coupled sucrose uptake system by a putative sucrose facilitator from Phaseolus vulgaris (PvSUF1). The resulting strains grew anaerobically on sucrose at specific growth rates of 0.09 ± 0.02 hâ1 (LmSPase) and 0.06 ± 0.01 hâ1 (PvSUF1, LmSPase). Overexpression of the yeast PGM2 gene, which encodes phosphoglucomutase, increased anaerobic growth rates on sucrose of these strains to 0.23 ± 0.01 hâ1 and 0.08 ± 0.00 hâ1, respectively. Determination of the biomass yield in anaerobic sucrose-limited chemostat cultures was used to assess the free-energy conservation of the engineered strains. Replacement of intracellular hydrolase with a phosphorylase increased the biomass yield on sucrose by 31%. Additional replacement of the native proton-coupled sucrose uptake system by PvSUF1 increased the anaerobic biomass yield by a further 8%, resulting in an overall increase of 41%. By experimentally demonstrating an energetic benefit of the combined engineering of disaccharide uptake and cleavage, this study represents a first step towards anaerobic production of compounds whose metabolic pathways currently do not conserve sufficient free-energy.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Metabolic Engineering - Volume 45, January 2018, Pages 121-133
Journal: Metabolic Engineering - Volume 45, January 2018, Pages 121-133
نویسندگان
Wesley Leoricy Marques, Robert Mans, Ryan K. Henderson, Eko Roy Marella, Jolanda ter Horst, Erik de Hulster, Bert Poolman, Jean-Marc Daran, Jack T. Pronk, Andreas K. Gombert, Antonius J.A. van Maris,