Adaptation of central metabolite pools to variations in growth rate and cultivation conditions in Saccharomyces cerevisiae.
Adaptation of central metabolite pools to variations in growth rate and cultivation conditions in Saccharomyces cerevisiae.
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DOI：
10.1186/s12934-021-01557-8
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发表时间：
2021-03-09
影响因子：
6.4
通讯作者：
Bruheim P
中科院分区：
文献类型：
--
作者：
Kumar K;Venkatraman V;Bruheim P
酿酒酵母是一种众所周知的基础生物学研究流行模型系统，可作为异源生产具有商业价值的小分子和蛋白质的宿主生物。中央代谢是提供构建模块和能量的核心，以支持正常情况下以及外源应激和强制异源蛋白质产生期间的生长和生存。在这里，我们对在不同碳源上分批培养和在营养限制葡萄糖恒化器中以不同生长速率生长酿酒酵母时的细胞内中心代谢物库分析进行了全面研究。最新版本的基于绝对定量质谱的代谢物分析方法适用于涵盖糖酵解和磷酸戊糖途径代谢物、三羧酸循环 (TCA)、完整氨基酸和脱氧/核苷磷酸池。在大多数测试条件下，谷氨酸、谷氨酰胺、丙氨酸和柠檬酸盐是四种最丰富的代谢物。氨基酸是主要的代谢物类别，尽管与其他代谢物类别相比，对于氮和磷酸盐限制的恒化剂观察到显着的相对减少。有趣的是，糖酵解和磷酸戊糖途径（PPP）代谢物在培养条件下显示出最大的变化，而核苷磷酸池则更稳定并且在更接近的浓度窗口内变化。葡萄糖和氮限制恒化剂的总体趋势是随着生长速率的增加，代谢物库增加。接下来，比较所选的恒化器参考生长速率（0.12 h−1，大约最大无限生长速率的四分之一）揭示了一个有趣的模式：与葡萄糖限制相比，几乎所有库的氮和磷酸盐限制条件都较低，除了 TCA 代谢物柠檬酸盐、异柠檬酸盐和 α-酮戊二酸盐。这项研究提供了新的知识——中央代谢如何适应不同的培养条件和生长速率，这对于扩大我们对细胞代谢的理解和代谢工程中改进表型的发展至关重要。在线版本包含可在 10.1186/s12934-021-01557-8 获取的补充材料。
Saccharomyces cerevisiae is a well-known popular model system for basic biological studies and serves as a host organism for the heterologous production of commercially interesting small molecules and proteins. The central metabolism is at the core to provide building blocks and energy to support growth and survival in normal situations as well as during exogenous stresses and forced heterologous protein production. Here, we present a comprehensive study of intracellular central metabolite pool profiling when growing S. cerevisiae on different carbon sources in batch cultivations and at different growth rates in nutrient-limited glucose chemostats. The latest versions of absolute quantitative mass spectrometry-based metabolite profiling methodology were applied to cover glycolytic and pentose phosphate pathway metabolites, tricarboxylic acid cycle (TCA), complete amino acid, and deoxy-/nucleoside phosphate pools. Glutamate, glutamine, alanine, and citrate were the four most abundant metabolites for most conditions tested. The amino acid is the dominant metabolite class even though a marked relative reduction compared to the other metabolite classes was observed for nitrogen and phosphate limited chemostats. Interestingly, glycolytic and pentose phosphate pathway (PPP) metabolites display the largest variation among the cultivation conditions while the nucleoside phosphate pools are more stable and vary within a closer concentration window. The overall trends for glucose and nitrogen-limited chemostats were increased metabolite pools with the increasing growth rate. Next, comparing the chosen chemostat reference growth rate (0.12 h−1, approximate one-fourth of maximal unlimited growth rate) illuminates an interesting pattern: almost all pools are lower in nitrogen and phosphate limited conditions compared to glucose limitation, except for the TCA metabolites citrate, isocitrate and α-ketoglutarate. This study provides new knowledge-how the central metabolism is adapting to various cultivations conditions and growth rates which is essential for expanding our understanding of cellular metabolism and the development of improved phenotypes in metabolic engineering. The online version contains supplementary material available at 10.1186/s12934-021-01557-8.
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影响因子：
5.7
作者：
Jung YH;Kim S;Yang J;Seo JH;Kim KH
通讯作者：
Kim KH
影响因子：
4.1
作者：
Edison AS;Hall RD;Junot C;Karp PD;Kurland IJ;Mistrik R;Reed LK;Saito K;Salek RM;Steinbeck C;Sumner LW;Viant MR
通讯作者：
Viant MR
影响因子：
4.8
作者：
Daran-Lapujade, P;Jansen, MLA;Pronk, JT
通讯作者：
Pronk, JT
影响因子：
8.4
作者：
Hameri, Tuure;Fengos, Georgios;Hatzimanikatis, Vassily
通讯作者：
Hatzimanikatis, Vassily
影响因子：
4.1
作者：
Martínez VS;Krömer JO
通讯作者：
Krömer JO