Metabolic engineering for aromatic amino acid biosynthesis pathway in Escherichia coli to acquire high-level biosynthesis of shikimic acid was reported. Knockout of aroL, ydiB genes and knock-in of T7-RNA-Polymerase gene which expression was controlled by L-arabinose based on the initial strain DH5α△ptsHIcrr (DHP), resulted in a series of shikimic acid-producing host strains. A series of tandem genes consisting of aroE, aroB, tktA, glk or aroFfbr were controlled by T7 promoter on plasmids were transformed into these host strains. According to the concentration of shikimic acid in shake-flask culture, all the engineered strains displayed high-potentiality compared to the control strain DHP, and the strain DHPYA-T7/pAOC-TGEFB synthesized the highest yield of 392 mg/L of shikimic acid. This study layed a strong foundation for constructing a high-level shikimic acid-producing engineered strain.