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以模型为指导的代谢重新连接,通过最大限度地减少碳损失来绕过pyruvate氧化,以合成pyruvate衍生物

Yun Zhang¹, Xueliang Wang^1,2, Christianah Odesanmi^1,2

¹State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

|February 5, 2024

概括

工程微生物绕过pyruvate氧化,以改善有价值的衍生物的生产. 这种新的代谢策略最大限度地减少了有氧生物过程中的碳损失,增强了工业化学合成.

科学领域:

代谢工程是代谢工程.
合成生物学合成生物学
微生物生物技术微生物生物技术

背景情况:

酸盐氧化对于微生物生长至关重要,但阻碍了酸盐衍生物的产生.
为了合成pyruvate衍生物,工程微生物宿主需要将新陈代谢与生长脱.

研究的目的:

通过最大限度地减少碳损失,开发一种微生物菌株来有效合成pyruvate衍生物.
在有氧条件下为葡萄糖培养的细菌建立一种新的代谢模式.

主要方法:

基因组规模的代谢建模和基于约束的流量平衡分析被用于菌株设计.
使用双突变通路和T7RNAP介导的合成小RNA来控制碳流量.
适应性实验室演变和多omics分析确定了关键的监管目标.

主要成果:

一个底盘菌株是使用glyoxylate和bifido道道构建的,绕过了pyruvate氧化.
改造品种产生了相当大的l-氨酸 (22.46 g/L),l-氨酸 (27.62 g/L) 和l-氨酸 (6.28 g/L) 的产量.
一种突变的异酸脱酶作为代谢开关,激活了glyoxylate突变.

结论:

建立了一个新的代谢模式,以最大限度地减少有氧生物过程中的碳损失.
工程微生物细胞工厂为生产酸衍生工业化学品提供了一个可持续的平台.
这项研究为在合成生物学中设计微生物细胞工厂提供了宝贵的见解.

关键词:

在T7RNAP中介的sRNA中.适应性实验室进化的演变.在cistronic图书馆里.基因组规模的代谢模型.进行多主题分析.皮鲁酸是一种酸盐.

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Fates of Pyruvate

Fates of Pyruvate

Pyruvate is the end product of glycolysis, where glucose is oxidized to pyruvate, simultaneously reducing NAD+ to NADH. Two molecules of ATP are also produced by substrate-level phosphorylation.
In aerobic organisms, pyruvate is metabolized via the citric acid cycle to produce reduced coenzymes NADH and FADH2. These coenzymes are then oxidized in the electron transport chain to produce ATP and, in the process, regenerate the NAD+ and FAD. As seen in some cell types and organisms, fermentation...

Pyruvate Oxidation

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After glycolysis, the charged pyruvate molecules enter the mitochondria via active transport and undergo three enzymatic reactions. These reactions ensure that pyruvate can enter the next metabolic pathway so that energy stored in the pyruvate molecules can be harnessed by the cells.
First, the enzyme pyruvate dehydrogenase removes the carboxyl group from pyruvate and releases it as carbon dioxide. The stripped molecule is then oxidized and releases electrons, which are then picked up by NAD+...

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Just like β-keto acids—which upon thermal decarboxylation form ketones—β-dicarboxylic acids undergo decarboxylation to generate monocarboxylic acids with the liberation of carbon dioxide.

What is Glycolysis?

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Overview
Cells make energy by breaking down macromolecules. Cellular respiration is the biochemical process that converts "food energy" (from the chemical bonds of macromolecules) into chemical energy in the form of adenosine triphosphate (ATP). The first step of this tightly regulated and intricate process is glycolysis. The word glycolysis originates from the Latin glyco (sugar) and lysis (breakdown). Glycolysis serves two main intracellular functions: generating ATP and generating...

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Loss of Carboxy Group as CO₂: Decarboxylation of β-Ketoacids

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Glycolysis: Pay-off Phase

Glycolysis: Pay-off Phase

So far, glycolysis has cost the cell two ATP molecules and produced two small, three-carbon sugar molecules. These molecules will proceed through the second half of the pathway, and sufficient energy will be extracted to pay back the two ATP molecules used as an initial investment and produce a profit for the cell of two additional ATP molecules and two even higher-energy NADH molecules.
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