无细胞代谢工程使脂肪酸能够选择性生物转化为增值化学品

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Yushi Liu , Wan-Qiu Liu , Shuhui Huang , Huiling Xu , Haofan Lu , Changzhu Wu , Jian Li
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引用次数: 4

摘要

脂肪酸衍生产品,如烷烃、脂肪醛和脂肪醇,在化学工业中有许多应用。这些产品主要由化石资源生产,但其生产过程往往不环保。虽然像大肠杆菌这样的微生物已经被改造成可以将脂肪酸转化为相应的产物,但由于低传质、重代谢负担和中间体/产物毒性,设计和优化细胞中的代谢途径以获得高产率是具有挑战性的。在这里,我们描述了一种基于大肠杆菌的无细胞蛋白质合成(CFPS)平台,用于体外将长链脂肪酸转化为具有产品选择性的增值化学品,在使用微生物生产系统时也可以避免上述问题。我们通过无细胞表达不同的酶和使用不同的条件(如光照和加热)来实现选择性的生物转化,以驱动生物催化产生不同的最终产物。具体而言,响应蓝光,无细胞表达的脂肪酸光羧化酶(CvFAP,一种光酶)能够以大约90%的转化率将脂肪酸转化为烷烃。当表达的酶转换为羧酸还原酶(CAR)时,脂肪酸被还原为相应的脂肪醛,然而,在无细胞系统中,脂肪醛可以通过内源性还原酶进一步还原为脂肪醇。通过使用热稳定的CAR和加热处理,内源性还原酶被失活,并且脂肪醛可以选择性地积累(在产物混合物中>97%)而不会过度还原为醇。总的来说,我们的无细胞平台提供了一种将脂肪酸转化为有价值化学物质的新策略,具有显著的操作灵活性、反应可控性和产物选择性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Cell-free metabolic engineering enables selective biotransformation of fatty acids to value-added chemicals

Cell-free metabolic engineering enables selective biotransformation of fatty acids to value-added chemicals

Fatty acid-derived products such as alkanes, fatty aldehydes, and fatty alcohols have many applications in the chemical industry. These products are predominately produced from fossil resources, but their production processes are often not environmentally friendly. While microbes like Escherichia coli have been engineered to convert fatty acids to corresponding products, the design and optimization of metabolic pathways in cells for high productivity is challenging due to low mass transfer, heavy metabolic burden, and intermediate/product toxicity. Here, we describe an E. coli-based cell-free protein synthesis (CFPS) platform for in vitro conversion of long-chain fatty acids to value-added chemicals with product selectivity, which can also avoid the above issues when using microbial production systems. We achieve the selective biotransformation by cell-free expression of different enzymes and the use of different conditions (e.g., light and heating) to drive the biocatalysis toward different final products. Specifically, in response to blue light, cell-free expressed fatty acid photodecarboxylase (CvFAP, a photoenzyme) was able to convert fatty acids to alkanes with approximately 90% conversion. When the expressed enzyme was switched to carboxylic acid reductase (CAR), fatty acids were reduced to corresponding fatty aldehydes, which, however, could be further reduced to fatty alcohols by endogenous reductases in the cell-free system. By using a thermostable CAR and a heating treatment, the endogenous reductases were deactivated and fatty aldehydes could be selectively accumulated (>97% in the product mixture) without over-reduction to alcohols. Overall, our cell-free platform provides a new strategy to convert fatty acids to valuable chemicals with notable properties of operation flexibility, reaction controllability, and product selectivity.

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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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