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ACS Synth Biol ; 13(7): 2081-2090, 2024 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-38607270

RESUMO

Ectoine is a compatible solute that functions as a cell protector from various stresses, protecting cells and stabilizing biomolecules, and is widely used in medicine, cosmetics, and biotechnology. Microbial fermentation has been widely used for the large-scale production of ectoine, and a number of fermentation strategies have been developed to increase the ectoine yield, reduce production costs, and simplify the production process. Here, Corynebacterium glutamicum was engineered for ectoine production by heterologous expression of the ectoine biosynthesis operon ectBAC gene from Halomonas elongata, and a series of genetic modifications were implemented. This included introducing the de3 gene from Escherichia coli BL21 (DE3) to express the T7 promoter, eliminating the lysine transporter protein lysE to limit lysine production, and performing a targeted mutation lysCS301Y on aspartate kinase to alleviate feedback inhibition of lysine. The new engineered strain Ect10 obtained an ectoine titer of 115.87 g/L in an optimized fed-batch fermentation, representing the highest ectoine production level in C. glutamicum and achieving the efficient production of ectoine in a low-salt environment.


Assuntos
Diamino Aminoácidos , Corynebacterium glutamicum , Escherichia coli , Fermentação , Halomonas , Engenharia Metabólica , Diamino Aminoácidos/biossíntese , Diamino Aminoácidos/metabolismo , Diamino Aminoácidos/genética , Corynebacterium glutamicum/genética , Corynebacterium glutamicum/metabolismo , Engenharia Metabólica/métodos , Halomonas/genética , Halomonas/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Lisina/metabolismo , Lisina/biossíntese , Regiões Promotoras Genéticas , Óperon/genética , Aspartato Quinase/genética , Aspartato Quinase/metabolismo , Sistemas de Transporte de Aminoácidos Básicos
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