In silico profiling of Escherichia coli and Saccharomyces cerevisiae as terpenoid factories.
In silico profiling of Escherichia coli and Saccharomyces cerevisiae as terpenoid factories.
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DOI：
10.1186/1475-2859-12-84
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发表时间：
2013-09-23
影响因子：
6.4
通讯作者：
Kayser O
中科院分区：
文献类型：
--
作者：
Gruchattka E;Hädicke O;Klamt S;Schütz V;Kayser O
异源微生物生产具有药用或工业价值的稀有植物萜类化合物正受到越来越多的关注，但萜类化合物的产量仍然较低。大肠杆菌和酿酒酵母是最广泛使用的异源宿主;然而，基于实验数据对两种宿主进行直接比较是困难的。因此，E. coli（通过1-脱氧-D-木酮糖-5-磷酸，DXP）和S.酿酒酵母（通过甲羟戊酸，MVA），通过基元模式分析在计算机上比较了各自宿主代谢的影响以及不同碳源的影响。本文重点研究了萜类化合物的前体--二磷酸异戊烯酯（IPP）的产率，旨在寻找提高萜类化合物产率的代谢工程新策略。从萜类途径的相应前体代谢物（DXP途径为丙酮酸和甘油醛-3-磷酸，MVA途径为乙酰辅酶A）开始，仅考虑碳化学计量，两种萜类途径在碳产率方面是相同的。然而，以葡萄糖为底物，如果考虑到所需前体的形成，MVA途径以高产率提供萜类化合物的潜力比DXP途径低，这是由于乙酰辅酶A形成中的碳损失。当考虑到所需的能量和还原当量时，这两种宿主中的最大产率进一步降低。此外，碳源（葡萄糖、木糖、乙醇或甘油）的选择对萜类化合物的产量有影响，其中不可发酵的碳源更有希望。两种宿主都缺乏用于高产量萜类化合物生产的能量和氧化还原当量，导致用于提高萜类化合物产量的新的过表达策略（异源酶/途径）。最后，使用强制生长与萜类化合物产率耦合的约束最小割集鉴定了几种敲除策略，所述萜类化合物产率高于迄今为止在科学文献中发表的任何产率。本研究首次提供了最主要的异源宿主E. coli和革兰氏阳性菌S.酿酒酵母作为萜类化合物的工厂，概述了几种有前途的代谢工程策略，为提高萜类化合物产量铺平了道路。
Heterologous microbial production of rare plant terpenoids of medicinal or industrial interest is attracting more and more attention but terpenoid yields are still low. Escherichia coli and Saccharomyces cerevisiae are the most widely used heterologous hosts; a direct comparison of both hosts based on experimental data is difficult though. Hence, the terpenoid pathways of E. coli (via 1-deoxy-D-xylulose 5-phosphate, DXP) and S. cerevisiae (via mevalonate, MVA), the impact of the respective hosts metabolism as well as the impact of different carbon sources were compared in silico by means of elementary mode analysis. The focus was set on the yield of isopentenyl diphosphate (IPP), the general terpenoid precursor, to identify new metabolic engineering strategies for an enhanced terpenoid yield. Starting from the respective precursor metabolites of the terpenoid pathways (pyruvate and glyceraldehyde-3-phosphate for the DXP pathway and acetyl-CoA for the MVA pathway) and considering only carbon stoichiometry, the two terpenoid pathways are identical with respect to carbon yield. However, with glucose as substrate, the MVA pathway has a lower potential to supply terpenoids in high yields than the DXP pathway if the formation of the required precursors is taken into account, due to the carbon loss in the formation of acetyl-CoA. This maximum yield is further reduced in both hosts when the required energy and reduction equivalents are considered. Moreover, the choice of carbon source (glucose, xylose, ethanol or glycerol) has an effect on terpenoid yield with non-fermentable carbon sources being more promising. Both hosts have deficiencies in energy and redox equivalents for high yield terpenoid production leading to new overexpression strategies (heterologous enzymes/pathways) for an enhanced terpenoid yield. Finally, several knockout strategies are identified using constrained minimal cut sets enforcing a coupling of growth to a terpenoid yield which is higher than any yield published in scientific literature so far. This study provides for the first time a comprehensive and detailed in silico comparison of the most prominent heterologous hosts E. coli and S. cerevisiae as terpenoid factories giving an overview on several promising metabolic engineering strategies paving the way for an enhanced terpenoid yield.
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影响因子：
8.4
作者：
Farhi, Moran;Marhevka, Elena;Vainstein, Alexander
通讯作者：
Vainstein, Alexander
DOI：
10.1126/science.1191652
发表时间：
2010-10-01
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Ajikumar PK;Xiao WH;Tyo KE;Wang Y;Simeon F;Leonard E;Mucha O;Phon TH;Pfeifer B;Stephanopoulos G
通讯作者：
Stephanopoulos G
影响因子：
12.3
作者：
Blank LM;Kuepfer L;Sauer U
通讯作者：
Sauer U
影响因子：
3.8
作者：
Förster, J;Gombert, AK;Nielsen, J
通讯作者：
Nielsen, J
影响因子：
4.4
作者：
Donald, KAG;Hampton, RY;Fritz, IB
通讯作者：
Fritz, IB