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MOESM1 of Metabolome analysis-based design and engineering of a metabolic pathway in Corynebacterium glutamicum to match rates of simultaneous utilization of d-glucose and l-arabinose

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posted on 2018-05-17, 05:00 authored by Hideo Kawaguchi, Kumiko Yoshihara, Kiyotaka Hara, Tomohisa Hasunuma, Chiaki Ogino, Akihiko Kondo
Additional file 1: Table S1. Specific growth rates and specific sugar consumption rates by recombinant strains and the wild-type strain of C. glutamicum ATCC 31831. Table S2. Relative expression levels of genes in the ara cluster and central metabolic pathway in aerobically grown wild-type and araR-deletion mutants with/without pyk overexpression (ΔaraR and ΔaraR/pCHpyk). Fig. S1. Retention time comparison of liquid chromatography–mass spectrometry/mass spectrometry total ion current chromatograms of oxaloacetate (OXA) in C. glutamicum cells grown aerobically in BT medium containing sugar mixture of d-glucose and l-arabinose (15 g/L each). The authentic standards of OXA at the concentration of 2, 10, and 20 μM (a, b, c) as well as their identified counterparts in C. glutamicum cells of the wild-type (d), and recombinant strains of 31831/pCHpyk (e) and ΔaraR/pCHpyk (f). OXA was identified based on retention time in chromatography and its mass spectrum. Concentration of OXA was determined with peak area of its multiple reaction monitoring (MRM) transition (131.0>87.1) in LC-MS/MS. Fig. S2. Mass spectra of identified oxaloacetate (OXA) in C. glutamicum cells grown aerobically in BT medium containing sugar mixture of d-glucose and l-arabinose (15 g/L each). The authentic standards of OXA at the concentration of 2, 10, and 20 μM (A) as well as their identified counterparts in C. glutamicum cells of the wild-type (d), and recombinant strains of 31831/pCHpyk (e) and ΔaraR/pCHpyk (f). OXA was identified based on retention time in chromatography and its mass spectrum acquired by the targeted multiple reaction monitoring (MRM) transition (131.0>87.1). Table S3. Oligonucleotides used for qPCR in this study.

Funding

Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Inter-disciplinary Research Areas (Innovative Bioproduction, Kobe) from the Ministry of Education, Culture, Sports, Science and Technology, Japan

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