Several studies on fermentative production of a vinyl monomer itaconic acid from hydrolyzed starch using Aspergillus terreus have been reported. Herein, we report itaconic acid production by Escherichia coli expressing recombinant alpha-amylase, using soluble starch as its sole carbon source. To express alpha-amylase in E. coli, we first constructed recombinant plasmids expressing alpha-amylases by using cell surface display technology derived from two amylolytic bacteria, Bacillus amyloliquefaciens NBRC 15535(T) and Streptococcus bovis NRIC 1535. The recombinant alpha-amylase from S. bovis (SBA) showed activity at 28 degrees C, which is the optimal temperature for production of itaconic acid, while alpha-amylase from B. amyloliquefaciens displayed no noticeable activity. E. coli cells expressing SBA produced 0.15 g/L itaconic acid after 69 h cultivation under pH-stat conditions, using 1% starch as the sole carbon source. In fact, E. coli cells expressing SBA had similar growth rates when grown in the presence of 1% glucose or starch, thereby highlighting the expression of an active alpha-amylase that enabled utilization of starch to produce itaconic acid in E. coli. (C) 2014, The Society for Biotechnology, Japan. All rights reserved.