Figure 3.

In Vivo and In Vitro Reconstitution of the MinA Mediated Assembly Line

(A) Schematic of the MinA-mediated assembly line for indigoidine biosynthesis.

(B) Engineered production of indigoidine in E. coli. Top, the target metabolite (blue) produced by related strains. Bottom, HPLC analysis of the target metabolite indigoidine produced by related E. coli strains. pET28a/indC, extracted metabolite of E. coli BAP1 containing indC (indigoidine synthetase gene from S. chromofuscus ATCC 49982) as positive control; pET28a/minA*, extracted metabolite of E. coli BAP1 containing minA*; pET28a, extracted metabolite of E. coli BAP1 containing pET28a as negative control.

(C) The extracted enzymatic products of the MinA reactions. Complete, the MinA reaction with all essential factors added; -Mg²⁺, the MinA reaction without adding exogenous Mg²⁺; -FMN, the MinA reaction without adding exogenous FMN; -L-Gln, the MinA reaction without L-glutamine added; -ATP, the MinA reaction without ATP added; -O₂, the MinA reaction under N₂ atmosphere.

(D) HPLC analysis of the related MinA reactions. The characteristic [M + H]⁺ ion of indigoidine was also indicated on this panel, and the information for related samples corresponds to that in panel C.

See also Figures S4 and S5; Tables S1 and S5.