| Autor: |
Amoah OJ; Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si 31460, Republic of Korea., Thapa SB; Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si 31460, Republic of Korea., Ma SY; Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si 31460, Republic of Korea., Nguyen HT; Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si 31460, Republic of Korea., Zakaria MM; Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si 31460, Republic of Korea., Sohng JK; Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si 31460, Republic of Korea.; Department of Pharmaceutical Engineering and Biotechnology, Sun Moon University, Asan-si 31460, Republic of Korea. |
| Abstrakt: |
Glucosylation is a well-known approach to improve the solubility, pharmacological, and biological properties of flavonoids, making flavonoid glucosides a target for large-scale biosynthesis. However, the low yield of products coupled with the requirement of expensive UDP-sugars limits the application of enzymatic systems for large-scale. C. glutamicum is a Gram-positive and generally regarded as safe (GRAS) bacteria frequently employed for the large-scale production of amino acids and bio-fuels. Due to the versatility of its cell factory system and its non-endotoxin producing properties, it has become an attractive system for the industrial-scale biosynthesis of alternate products. Here, we explored the cell factory of C. glutamicum for efficient glucosylation of flavonoids using apigenin as a model flavonoid, with the heterologous expression of a promiscuous glycosyltransferase, YdhE from Bacillus licheniformis and the endogenous overexpression of C. glutamicum genes galU1 encoding UDP-glucose pyrophosphorylase and pgm encoding phosphoglucomutase involved in the synthesis of UDP-glucose to create a C. glutamicum cell factory system capable of efficiently glucosylation apigenin with a high yield of glucosides production. Consequently, the production of various apigenin glucosides was controlled under different temperatures yielding almost 4.2 mM of APG1(apigenin-4'-O-β-glucoside) at 25°C, and 0.6 mM of APG2 (apigenin-7-O-β-glucoside), 1.7 mM of APG3 (apigenin-4',7-O-β-diglucoside) and 2.1 mM of APG4 (apigenin-4',5-O-β-diglucoside) after 40 h of incubation with the supplementation of 5 mM of apigenin and 37°C. The cost-effective developed system could be used to modify a wide range of plant secondary metabolites with increased pharmacokinetic activities on a large scale without the use of expensive UDP-sugars. |
