caa a22 4500 605498997 CHVBK 20210128100549.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6303-5 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6303-5 Promoters inducible by aromatic amino acids and γ-aminobutyrate (GABA) for metabolic engineering applications in Saccharomyces cerevisiae [Elektronische Daten] [Sujin Kim, Kyusung Lee, Sang-Jeong Bae, Ji-Sook Hahn] A wide range of promoters with different strengths and regulatory mechanisms are valuable tools in metabolic engineering and synthetic biology. While there are many constitutive promoters available, the number of inducible promoters is still limited for pathway engineering in Saccharomyces cerevisiae. Here, we constructed aromatic amino-acid-inducible promoters based on the binding sites of Aro80 transcription factor, which is involved in the catabolism of aromatic amino acids through transcriptional activation of ARO9 and ARO10 genes in response to aromatic amino acids. A dynamic range of tryptophan-inducible promoter strengths can be obtained by modulating the number of Aro80 binding sites, plasmid copy numbers, and tryptophan concentrations. Using low and high copy number plasmid vectors and different tryptophan concentrations, a 29-fold range of fluorescence intensities of enhanced green fluorescent protein (EGFP) reporter could be achieved from a synthetic U4C ARO9 promoter, which is composed of four repeats of Aro80 binding half site (CCG) and ARO9 core promoter element. The U4C ARO9 promoter was applied to express alsS and alsD genes from Bacillus subtilis for acetoin production in S. cerevisiae, resulting in a gradual increase in acetoin titers depending on tryptophan concentrations. Furthermore, we demonstrated that γ-aminobutyrate (GABA)-inducible UGA4 promoter, regulated by Uga3, can also be used in metabolic engineering as a dose-dependent inducible promoter. The wide range of controllable expression levels provided by these tryptophan- and GABA-inducible promoters might contribute to fine-tuning gene expression levels and timing for the optimization of pathways in metabolic engineering. Springer-Verlag Berlin Heidelberg, 2015 Acetoin nationallicence Aro80 nationallicence Aromatic amino acid nationallicence GABA nationallicence Inducible promoter nationallicence Saccharomyces cerevisiae nationallicence Kim Sujin School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, 151-744, Seoul, Republic of Korea aut Lee Kyusung School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, 151-744, Seoul, Republic of Korea aut Bae Sang-Jeong School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, 151-744, Seoul, Republic of Korea aut Hahn Ji-Sook School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, 151-744, Seoul, Republic of Korea aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2705-2714 0175-7598 99:6<2705 2015 99 253 https://doi.org/10.1007/s00253-014-6303-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6303-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kim Sujin School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, 151-744, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Lee Kyusung School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, 151-744, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Bae Sang-Jeong School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, 151-744, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Hahn Ji-Sook School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, 151-744, Seoul, Republic of Korea aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2705-2714 0175-7598 99:6<2705 2015 99 253