Construction of a Vitreoscilla Hemoglobin Promoter-Based Tunable Expression System for Corynebacterium glutamicum
| Autor: | Seo Young Jo, Jong Hyun Choi, Bong Keun Song, Si Jae Park, Dae-Hee Lee, Mi Na Rhie, Binna Lee, Kyoung Hee Kang, Hee Taek Kim, Kei Anne Baritugo, Jeong Chan Joo, Jae Jun Song, Tae Uk Khang |
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| Rok vydání: | 2018 |
| Předmět: |
Pvgb
02 engineering and technology lcsh:Chemical technology 010402 general chemistry 01 natural sciences Catalysis Corynebacterium glutamicum Green fluorescent protein law.invention lcsh:Chemistry Metabolic engineering law lcsh:TP1-1185 Physical and Theoretical Chemistry Vitreoscilla vgb chemistry.chemical_classification Expression vector Strain (chemistry) biology Chemistry tunable expression system 021001 nanoscience & nanotechnology biology.organism_classification 0104 chemical sciences Enzyme lcsh:QD1-999 Biochemistry Recombinant DNA bacteria synthetic biology 0210 nano-technology expression vectors |
| Zdroj: | Catalysts Volume 8 Issue 11 Catalysts, Vol 8, Iss 11, p 561 (2018) |
| ISSN: | 2073-4344 |
| DOI: | 10.3390/catal8110561 |
| Popis: | Corynebacterium glutamicum is an industrial strain used for the production of valuable chemicals such as L-lysine and L-glutamate. Although C. glutamicum has various industrial applications, a limited number of tunable systems are available to engineer it for efficient production of platform chemicals. Therefore, in this study, we developed a novel tunable promoter system based on repeats of the Vitreoscilla hemoglobin promoter (Pvgb). Tunable expression of green fluorescent protein (GFP) was investigated under one, four, and eight repeats of Pvgb (Pvgb, Pvgb4, and Pvgb8). The intensity of fluorescence in recombinant C. glutamicum strains increased as the number of Pvgb increased from single to eight (Pvgb8) repeats. Furthermore, we demonstrated the application of the new Pvgb promoter-based vector system as a platform for metabolic engineering of C. glutamicum by investigating 5-aminovaleric acid (5-AVA) and gamma-aminobutyric acid (GABA) production in several C. glutamicum strains. The profile of 5-AVA and GABA production by the recombinant strains were evaluated to investigate the tunable expression of key enzymes such as DavBA and GadBmut. We observed that 5-AVA and GABA production by the recombinant strains increased as the number of Pvgb used for the expression of key proteins increased. The recombinant C. glutamicum strain expressing DavBA could produce higher amounts of 5-AVA under the control of Pvgb8 (3.69 ± 0.07 g/L) than the one under the control of Pvgb (3.43 ± 0.10 g/L). The average gamma-aminobutyric acid production also increased in all the tested strains as the number of Pvgb used for GadBmut expression increased from single (4.81⁻5.31 g/L) to eight repeats (4.94⁻5.58 g/L). |
| Databáze: | OpenAIRE |
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