Metabolic engineering of Escherichia coli BL21(DE3) for efficient production of indigo using Methylophaga aminisulfidivorans flavin-containing monooxygenase

IF 4.1 3区工程技术 Q2 CHEMISTRY, APPLIED

Dyes and Pigments Pub Date : 2025-01-27 DOI:10.1016/j.dyepig.2025.112682

Rui Lu , Roulin Chen , Zeyu Li , Heng Hu , Zhimeng Wu , Yingying Zhu , Wanmeng Mu

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Abstract

Biological production of indigo can be achieved by enzymatic hydroxylation of indole, the degradation product of tryptophan. Many oxygenases have been identified to be capable of producing indigo from indole, among which flavin-containing monooxygenase (FMO) is the most extensively studied. In this study, Escherichia coli BL21 (DE3) was metabolically engineered to efficiently produce indigo using FMO. Five reported indigo-producing FMOs and five homologous human FMOs were selected to evaluate the in vivo indigo production abilities, in which Methylophaga aminisulfidivorans FMO showed the highest indigo titer. Various metabolic engineering strategies were performed to stepwise enhance the indigo titer, including strengthening tryptophan transportation, blocking tryptophan synthesis-related competitive pathways, regulating key metabolic genes for tryptophan synthesis. When supplemented with tryptophan, the final engineered strain produced 532 and 1492 mg/L of indigo by shake-flask and fed-batch cultivation, respectively.

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来源期刊

Dyes and Pigments 工程技术-材料科学：纺织

CiteScore

8.20

自引率

13.30%

发文量

933

审稿时长

33 days

期刊介绍： Dyes and Pigments covers the scientific and technical aspects of the chemistry and physics of dyes, pigments and their intermediates. Emphasis is placed on the properties of the colouring matters themselves rather than on their applications or the system in which they may be applied. Thus the journal accepts research and review papers on the synthesis of dyes, pigments and intermediates, their physical or chemical properties, e.g. spectroscopic, surface, solution or solid state characteristics, the physical aspects of their preparation, e.g. precipitation, nucleation and growth, crystal formation, liquid crystalline characteristics, their photochemical, ecological or biological properties and the relationship between colour and chemical constitution. However, papers are considered which deal with the more fundamental aspects of colourant application and of the interactions of colourants with substrates or media. The journal will interest a wide variety of workers in a range of disciplines whose work involves dyes, pigments and their intermediates, and provides a platform for investigators with common interests but diverse fields of activity such as cosmetics, reprographics, dye and pigment synthesis, medical research, polymers, etc.