Daoan Wang , Jiamin Chen , Yang Wang , Guocheng Du , Zhen Kang
{"title":"Engineering Escherichia coli for high-yield production of ectoine","authors":"Daoan Wang , Jiamin Chen , Yang Wang , Guocheng Du , Zhen Kang","doi":"10.1016/j.gce.2021.09.002","DOIUrl":null,"url":null,"abstract":"<div><p>Ectoine is a natural macromolecule protector and synthesized by some extremophiles. It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and cosmetics. To meet its growing commercial demands, we engineered <em>Escherichia coli</em> strains for the high-yield production of ectoine. The <em>ectABC</em> gene cluster from the native ectoine producer <em>Halomonas elongata</em> was introduced into different <em>Escherichia coli (E. Coil)</em> strains <em>via</em> plasmids and 0.8 g L<sup>-1</sup> of ectoine was produced in flask cultures by engineered <em>E. coli</em> BL21 (DE3). Subsequently, we designed the ribosome-binding sites of the gene cluster to fine-tune the expressions of genes <em>ectA</em>, <em>ectB</em>, and <em>ectC</em>, which increased the ectoine yield to 1.6 g L<sup>-1</sup>. After further combinatorial overexpression of <em>Corynebacterium glutamicum</em> aspartate kinase mutant (G1A, C932T) and the <em>H. elongate</em> aspartate-semialdehyde dehydrogenase to increase the supply of the precursor, the titer of ectoine reached to 5.5 g L<sup>-1</sup> in flask cultures. Finally, the engineered strain produced 60.7 g L<sup>-1</sup> ectoine in fed-batch cultures with a conversion rate of 0.25 g/g glucose.</p></div>","PeriodicalId":66474,"journal":{"name":"Green Chemical Engineering","volume":null,"pages":null},"PeriodicalIF":9.1000,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"9","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Green Chemical Engineering","FirstCategoryId":"1089","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2666952821000662","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ENGINEERING, CHEMICAL","Score":null,"Total":0}
引用次数: 9
Abstract
Ectoine is a natural macromolecule protector and synthesized by some extremophiles. It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and cosmetics. To meet its growing commercial demands, we engineered Escherichia coli strains for the high-yield production of ectoine. The ectABC gene cluster from the native ectoine producer Halomonas elongata was introduced into different Escherichia coli (E. Coil) strains via plasmids and 0.8 g L-1 of ectoine was produced in flask cultures by engineered E. coli BL21 (DE3). Subsequently, we designed the ribosome-binding sites of the gene cluster to fine-tune the expressions of genes ectA, ectB, and ectC, which increased the ectoine yield to 1.6 g L-1. After further combinatorial overexpression of Corynebacterium glutamicum aspartate kinase mutant (G1A, C932T) and the H. elongate aspartate-semialdehyde dehydrogenase to increase the supply of the precursor, the titer of ectoine reached to 5.5 g L-1 in flask cultures. Finally, the engineered strain produced 60.7 g L-1 ectoine in fed-batch cultures with a conversion rate of 0.25 g/g glucose.