在Komagataella phaffii中设计合成β-alanine通路,将甲醇转化为3-基酸
Sílvia Àvila-Cabré1, Míriam Pérez-Trujillo2, Joan Albiol1
1Department of Chemical, Biological and Environmental Engineering, Universitat Autònoma de Barcelona, Bellaterra, Catalonia, Spain.
Microbial cell factories
|November 18, 2023
概括
这项研究确立了Komagataella phaffii作为一种从甲醇中生产3-基酸 (3-HP) 的细胞工厂. 工程酵母菌株取得了高产量,证明了可持续化学生产的潜力.
科学领域:
- 合成生物学和代谢工程合成生物学和代谢工程
- 工业生物技术 工业生物技术
- 可再生化工生产的可再生化工生产
背景情况:
- 甲醇是化学合成的有希望的可再生碳来源.
- 科马加泰拉 (Komagataella phaffii) (Pichia pastoris) 是一种具有细胞工厂潜力的甲基转化酵母.
- 3-基酸 (3-HP) 是生物聚合物和酸的关键平台化学物质.
研究的目的:
- 通过合成代谢途径在Komagataella phaffii中建立3-HP生产.
- 设计K. phaffii以从甲醇中增强生产3-HP.
- 评估K. phaffii作为从C1原料生产有机酸的平台的潜力.
主要方法:
- 三个基因 (panD, yhxA, ydfG) 的异质表达,用于构建K. phaffii.中的合成β-氨酸通路.
- 菌株工程包括基因复制和促进物选择以优化3HP生产.
- 在与过程相关的条件下进行料批量培养实验,以确定生产性能.
- 通过工程进一步改善菌株,以增加NADPH的可用性.
主要成果:
- 通过合成β-氨酸途径,成功地在K. phaffii中建立了3-HP生产.
- 在小规模培养中达到1.0g/L的最终3HP标位,在菌株优化时达到1.2g/L.
- 料批发发酵产生了21.4 g/L的3-HP,产量为0.15 g/g,生产率为0.48 g/L/h.
- 工程菌株显示甲醇消耗增加和特定生产率.
结论:
- 证明了使用K. phaffii作为从甲醇生产3HP的细胞工厂的可行性.
- 通过β-氨酸途径实现了通过β-氨酸途径为3HP生产报告的最高体积生产率.
- 强调了K. phaffii在从可再生C1原料中可持续生产有机酸的潜力.
相关概念视频
01:29Amino Acid Biosynthetic Pathways
17
Amino acid biosynthesis is essential for cell growth, protein synthesis, and metabolic regulation. Cells generate essential and non-essential amino acids from metabolic intermediates to sustain vital biological functions. These intermediates originate from key metabolic pathways: glycolysis, the tricarboxylic acid (TCA) cycle, and the pentose phosphate pathway. Important precursors include α-ketoglutarate, pyruvate, oxaloacetate, phosphoenolpyruvate, and erythrose-4-phosphate, which...
17
01:07Alkylation of β-Ketoester Enolates: Acetoacetic Ester Synthesis
3.3K
Acetoacetic ester synthesis is a method to obtain ketones from alkyl halides and β-keto esters. The reaction occurs in the presence of an alkoxide base that abstracts the acidic proton of the β-keto esters. The step results in an enolate ion which is doubly stabilized. The enolate then reacts with an alkyl halide via the SN2 process to produce an alkylated ester intermediate with a new C–C bond. The hydrolysis of the intermediate, followed by acidification, results in an...
3.3K
01:28Carbon-dioxide Fixation
19
Carbon dioxide fixation in prokaryotes enables the assimilation of inorganic carbon into organic molecules, supporting biosynthetic pathways, sustaining ecosystems, and contributing to the global carbon cycle. It also has industrial applications in carbon capture and bioproduct synthesis. Autotrophic organisms rely on this process to utilize CO₂ as a carbon source in diverse environments.The Calvin CycleThe Calvin cycle is the most widespread carbon fixation mechanism, primarily used by...
19
01:14Alkylation of β-Diester Enolates: Malonic Ester Synthesis
3.4K
Malonic ester synthesis is a method to obtain α substituted carboxylic acids from ꞵ-diesters such as diethyl malonate and alkyl halides.
3.4K
01:35Loss of Carboxy Group as CO2: Decarboxylation of Malonic Acid Derivatives
2.0K
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.
2.0K
01:22Reactions of Aldehydes and Ketones: Baeyer–Villiger Oxidation
4.2K
Baeyer–Villiger oxidation converts aldehydes to carboxylic acids and ketones to esters. The reaction uses peroxy acids or peracids and is often catalyzed by acid. The reaction is named after its pioneers, Adolf von Baeyer and Victor Villiger. The reaction is achieved by a wide range of peracids such as m-chloroperoxybenzoic acid (mCPBA), perbenzoic acid (C6H5COOOH), peracetic acid (CH3COOOH), hydrogen peroxide (H2O2), and tert-butyl hydroperoxide (t-BuOOH).
The carbonyl center is...
The carbonyl center is...
4.2K