Engineering of itaconic acid pathway via co-localization of CadA and AcnA in recombinant Escherichia coli.

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-05-29 DOI:10.1007/s10529-024-03496-x
Kim-Ngan T Tran, Jaehoon Jeong, Soon Ho Hong
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Abstract

Itaconic acid is an excellent polymeric precursor with a wide range of industrial applications. The efficient production of itaconate from various renewable substrates was demonstrated by engineered Escherichia coli. However, limitation in the itaconic acid precursor supply was revealed by finding out the key intermediate of the tricarboxylic acid in the itaconic acid pathway. Efforts of enhancing the cis-aconitate flux and preserving the isocitrate pool to increase itaconic acid productivity are required. In this study, we introduce a synthetic protein scaffold system between CadA and AcnA to physically combine the two enzymes. Through the introduction of a synthetic protein scaffold, 2.1 g L-1 of itaconic acid was produced at pH 7 and 37 °C. By fermentation, 20.1 g L-1 for 48 h of itaconic acid was produced with a yield of 0.34 g g-1 glycerol. These results suggest that carbon flux was successfully increased itaconic acid productivity.

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通过 CadA 和 AcnA 在重组大肠杆菌中的共定位改造衣康酸途径。
衣康酸是一种优良的聚合物前体,具有广泛的工业用途。经改造的大肠杆菌证明,可以从各种可再生底物中高效生产衣康酸。然而,通过发现衣康酸途径中三羧酸的关键中间体,发现了衣康酸前体供应的局限性。因此,需要努力提高顺式-乌头酸通量并保留异柠檬酸池,以提高衣康酸的生产率。在这项研究中,我们在 CadA 和 AcnA 之间引入了一个合成蛋白支架系统,将这两种酶进行物理结合。通过引入合成蛋白支架,在 pH 值为 7、温度为 37 ℃ 的条件下产生了 2.1 g L-1 的衣康酸。通过发酵,48 小时内产生了 20.1 g L-1 的衣康酸,甘油产量为 0.34 g-1。这些结果表明,碳通量成功地提高了衣康酸的生产率。
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来源期刊
Biotechnology Letters
Biotechnology Letters 工程技术-生物工程与应用微生物
CiteScore
5.90
自引率
3.70%
发文量
108
审稿时长
1.2 months
期刊介绍: Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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