Kozaeva Ekaterina, Nieto-Domínguez Manuel, Tang Kent Kang Yong, Stammnitz Maximilian, Nikel Pablo Iván
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, 2800, Denmark.
Centre for Genomic Regulation, Barcelona 08003, Spain.
ACS Synth Biol. 2025 Jan 17;14(1):257-272. doi: 10.1021/acssynbio.4c00700. Epub 2025 Jan 3.
Methyl ketones, key building blocks widely used in diverse industrial applications, largely depend on oil-derived chemical methods for their production. Here, we investigated biobased production alternatives for short-chain ketones, adapting the solvent-tolerant soil bacterium as a host for ketone biosynthesis either by whole-cell biocatalysis or using engineered minicells, chromosome-free bacterial vesicles. Organic acids (acetate, propanoate and butanoate) were selected as the main carbon substrate to drive the biosynthesis of acetone, butanone and 2-pentanone. Pathway optimization identified efficient enzyme variants from and , tested with both constitutive and inducible expression of the cognate genes. By implementing these optimized pathways in minicells, which can be prepared through a simple three-step purification protocol, the feedstock was converted into the target short-chain methyl ketones. These results highlight the value of combining morphology and pathway engineering of noncanonical bacterial hosts to establish alternative bioprocesses for toxic chemicals that are difficult to produce by conventional approaches.
甲基酮是广泛应用于各种工业领域的关键构件,其生产在很大程度上依赖于石油衍生的化学方法。在此,我们研究了短链酮的生物基生产替代方案,将耐溶剂土壤细菌作为酮生物合成的宿主,通过全细胞生物催化或使用工程化微细胞(无染色体细菌囊泡)来实现。选择有机酸(乙酸盐、丙酸盐和丁酸盐)作为主要碳底物,以驱动丙酮、丁酮和2-戊酮的生物合成。通过途径优化,从[具体来源]中鉴定出高效的酶变体,并对同源基因的组成型和诱导型表达进行了测试。通过在可通过简单的三步纯化方案制备的微细胞中实施这些优化途径,原料被转化为目标短链甲基酮。这些结果突出了结合非典型细菌宿主的形态学和途径工程来建立传统方法难以生产的有毒化学品的替代生物工艺的价值。