We engineered a Corynebacterium glutamicum strain displaying α-amylase from Streptococcus bovis 148 (AmyA) on its cell surface to produce amino acids directly from starch. We used PgsA from Bacillus subtilis as an anchor protein, and the N-terminus of α-amylase was fused to the PgsA. The genes of the fusion protein were integrated into the homoserine dehydrogenase gene locus on the chromosome by homologous recombination. l-Lysine fermentation was carried out using C. glutamicum displaying AmyA in the growth medium containing 50 g/l soluble starch as the sole carbon source. We performed l-lysine fermentation at various temperatures (30–40°C) and pHs (6.0–7.0), as the optimal temperatures and pHs of AmyA and C. glutamicum differ significantly. The highest l-lysine yield was recorded at 30°C and pH 7.0. The amount of soluble starch was reduced to 18.29 g/l, and 6.04 g/l l-lysine was produced in 24 h. The l-lysine yield obtained using soluble starch as the sole carbon source was higher than that using glucose as the sole carbon source after 24 h when the same amount of substrates was added. The results shown in the current study demonstrate that C. glutamicum displaying α-amylase has a potential to directly convert soluble starch to amino acids.
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We would like to express our gratitude to Dr. Masahiro Hatsu and Mr. Takashi Mimitsuka of New Frontiers Research Laboratories, Toray Industries for providing the plasmid pTM44 and for advices on the experimental procedures, and to Dr. Moon-Hee Sung and Dr. Tomomitsu Sewaki of Bioleaders Japan Corporation for providing the PgsA gene.
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Tateno, T., Fukuda, H. & Kondo, A. Production of l-Lysine from starch by Corynebacterium glutamicum displaying α-amylase on its cell surface. Appl Microbiol Biotechnol 74, 1213–1220 (2007). https://doi.org/10.1007/s00253-006-0766-y
- Sole Carbon Source
- Soluble Starch
- Corynebacterium Glutamicum
- Cell Surface Display