title: Production of itaconic acid in Escherichia coli expressing recombinant α-amylase using starch as substrate.
abstract: 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 α-amylase, using soluble starch as its sole carbon source. To express α-amylase in E. coli, we first constructed recombinant plasmids expressing α-amylases by using cell surface display technology derived from two amylolytic bacteria, Bacillus amyloliquefaciens NBRC 15535(T) and Streptococcus bovis NRIC 1535. The recombinant α-amylase from S. bovis (SBA) showed activity at 28°C, which is the optimal temperature for production of itaconic acid, while α-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 α-amylase that enabled utilization of starch to produce itaconic acid in E. coli.
authors.affiliation: Department of Biobased Materials Science, Kyoto Institute of Technology, 1 Hashigami-cho, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Shusuke
authors.initials: S
authors.lastname: Okamoto
authors.affiliation: Department of Biobased Materials Science, Kyoto Institute of Technology, 1 Hashigami-cho, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Taejun
authors.initials: T
authors.lastname: Chin
authors.affiliation: Department of Biobased Materials Science, Kyoto Institute of Technology, 1 Hashigami-cho, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Nagata
authors.affiliation: Faculty of Education, Shimane University, 1060 Nishikawatsu-cho, Matsue, Shimane 690-8504, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Takahashi
authors.affiliation: Department of Biobased Materials Science, Kyoto Institute of Technology, 1 Hashigami-cho, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Ohara
authors.affiliation: Department of Biobased Materials Science, Kyoto Institute of Technology, 1 Hashigami-cho, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan. Electronic address: aso@kit.ac.jp.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Aso
mesh_terms: D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D013213:Starch / Q000378:metabolism*; D016985:Streptococcus bovis / Q000201:enzymology / Q000235:genetics; D013386:Succinates / Q000378:metabolism*; D013696:Temperature; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D013386:Succinates; D013213:Starch; D000516:alpha-Amylases; D005947:Glucose; C005229:itaconic acid
keywords: Bacillus amyloliquefaciens; Escherichia coli; Itaconic acid; Starch; Streptococcus bovis; α-Amylase