High-Level Production of the Natural Blue Pigment Indigoidine from Metabolically Engineered Corynebacterium glutamicum for Sustainable Fabric Dyes

Abstract

The textile industry has caused severe water pollution by using many toxic chemicals for producing fabric dyes. In response to this problem, indigoidine has attracted attention as an alternative natural blue dye, but it is necessary to achieve a high-level production to compete with synthetic blue dyes. Here we report a metabolically engineered Corynebacterium glutamicum capable of producing indigoidine to a high concentration with high productivity. First, the blue-pigment indigoidine synthetase (bpsA) gene from Streptomyces lavendulae was expressed in C. glutamicum, which carries strong fluxes toward L-glutamate, a precursor of indigoidine. Production performance of this base strain, already producing 7.3 ± 0.3 g/L indigoidine from the flask, was further improved by streamlining the intracellular supply of the precursors L-glutamate and L-glutamine, strengthening the phosphotransferase system-independent glucose uptake system, channeling carbon fluxes from glycolysis to the tricarboxylic acid (TCA) cycle, and minimizing byproducts formation. Fedbatch fermentation of the final strain BIRU11 produced 49.30 g/L indigoidine with a productivity of 0.96 g/L/h, the highest titer and productivity to date. Finally, indigoidine from the fed-batch fermentation of the BIRU11 strain was used to dye white cotton fabrics to examine its color and performance. This study demonstrates the potential of producing fabric dyes in a sustainable manner by using a metabolically engineered bacterium.

Generation of Pseudomonas putida KT2440 Strains with Efficient Utilization of Xylose and Galactose via Adaptive Laboratory Evolution

“…In particular, the Δ gcd strains were included, given that the deletion did not significantly affect the growth rates on glucose (Figure S3). For this demonstration, we introduced the production pathway of indigoidine, which is a natural pigment and has industrial interest, ,− in evolved isolates (Figure A). It was previously shown that indigoidine can be produced by the heterologous expression of bpsA encoding blue pigment synthetase A from Streptomyces lavendulae and sfp encoding 4′-phosphopantetheinyl transferase from Bacillus subtilis . , We expressed the two genes under the arabinose-inducible promoter and integrated them into the genome (see Methods).…”

De Novo Biosynthesis and Whole-Cell Catalytic Production of 2-Acetamidophenol in Escherichia coli

“…Numerous studies have shown that fed-batch fermentation is an effective method to increase the accumulation of the target products. For instance, fed-batch fermentation of metabolically engineered Corynebacterium glutamicum produced indigoidine, 37 engineered E. coli yielded L-valine by two-stage fed-batch fermentation, 38 and aromatic compounds phenol and p-amino-L-phenylalanine were also produced by engineered E. coli in fed-batch cultivation. 39,40 When the cell density reached 20 (OD 600 ), the recombinant strains were induced by IPTG.…”

Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites

“…In a similar effort to improve the synthesis of indogoidine, the S. lavendulae bpsA gene has been transferred to the industrial production strains Corynebacterium glutamicum and Rhodosporidium toruloides. When combined with metabolic engineering and the optimization of fermentation strategies, this achieved indigoidine yields of 49 and 86 g per L, respectively (Wehrs et al, 2019;Ghiffary et al, 2021). Indigoidine has also been produced by cell-free biosynthesis in the commercial E. colibased PURExpress system (Siebels et al, 2020).…”