Enhanced production of L-phenylalanine in Corynebacterium glutamicum due to the introduction of Escherichia coli wild-type gene aroH

• Published in: Journal of industrial microbiology & biotechnology Vol. 40; no. 6; pp. 643 - 651
• Main Authors: Zhang, Chuanzhi, Zhang, Junli, Kang, Zhen, Du, Guocheng, Yu, Xiaobin, Wang, Tianwen, Chen, Jian
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.06.2013; Springer; Oxford University Press
• Subjects: Alkyl and Aryl Transferases - genetics; Alkyl and Aryl Transferases - metabolism; Amino acids; Analysis; Bacteria; biochemical pathways; Biochemistry; Bioengineering; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biosynthesis; Biosynthetic Pathways - genetics; Biotechnology; Chemical synthesis; Cloning; Corynebacterium glutamicum; Corynebacterium glutamicum - genetics; Corynebacterium glutamicum - metabolism; E coli; Engineering; Enzymes; Escherichia coli; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Feedback; Feedback, Physiological; Fundamental and applied biological sciences. Psychology; Gene Expression; gene expression regulation; gene overexpression; Genes; Genetic Engineering; Inorganic Chemistry; Laboratories; Life Sciences; Metabolic Engineering; Metabolic Engineering and Synthetic Biology; Metabolism; Microbiology; Multienzyme Complexes - genetics; Multienzyme Complexes - metabolism; Phenylalanine - biosynthesis; Plasmids; Prephenate Dehydratase - genetics; Prephenate Dehydratase - metabolism; Studies; phenylalanine; Metabolic engineering; L-phenylalanine; Gene; Phenylalanine; Corynebacterium glutamicum; Escherichia coli; Production; aroH; Bacteria; Actinomycetes; Corynebacteriaceae; Enterobacteriaceae
• ISSN: 1367-5435; 1476-5535
• DOI: 10.1007/s10295-013-1262-x

Metabolic engineering is a powerful tool which has been widely used for producing valuable products. For improving L-phenylalanine (L-Phe) accumulation in Corynebacterium glutamicum, we have investigated the target genes involved in the biosynthetic pathways. The genes involved in the biosynthesis of L-Phe were found to be strictly regulated genes by feedback inhibition. As a result, overexpression of the native wild-type genes aroF, aroG or pheA resulted in a slight increase of L-Phe. In contrast, overexpression of aroF ʷᵗ or pheA ᶠᵇʳ from E. coli significantly increased L-Phe production. Co-overexpression of aroF ʷᵗ and pheA ᶠᵇʳ improved the titer of L-Phe to 4.46 ± 0.06 g l⁻¹. To further analyze the target enzymes in the aromatic amino acid synthesis pathway between C. glutamicum and E. coli, the wild-type gene aroH from E. coli was overexpressed and evaluated in C. glutamicum. As predicted, upregulation of the wild-type gene aroH resulted in a remarkable increase of L-Phe production. Co-overexpression of the mutated pheA ᶠᵇʳ and the wild-type gene aroH resulted in the production of L-Phe up to 4.64 ± 0.09 g l⁻¹. Based on these results we conclude that the wild-type gene aroH from E. coli is an appropriate target gene for pathway engineering in C. glutamicum for the production of aromatic amino acids.