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- 저자 : ( Obed Jackson Amoah ) (검색결과 3 건)
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N-glucosylation in Corynebacterium glutamicum with YdhE from Bacillus lichenformis
Obed Jackson Amoah,Hue Thi Nguyen,Su Yeong Ma,Jae Kyung Sohng 한국당과학회 2022 한국당과학회 학술대회 Vol.2022 No.07
A system was developed to successfully biosynthesize glucoside derivatives of therapeutic and relevant N-linked compounds in the generally regarded as safe (GRAS) Corynebacterium glutamicum. Using glucose, as the sole carbon source, YdhE, which is a promiscuous glycosyltransferase from Bacillus lichenformis and has been introduced in the Corynebacterium glutamicum cell factory system, was able to biosynthesize most of the N-linked compounds to their rare glucosides. Optimal conditions such as IPTG concentrations and induction time, Temperature, Substrate and glucose concentration, and Production time on the In Vivo conversion were successfully developed. This yielded almost 85% of glucosylated butyl-4-aminobenzoate glucoside after only 10 hours post substrate induction. The developed system uses recombinant Corynebacterium glutamicum to achieve In Vivo glucosylation using cheap glucose as the main source of UDP-sugar moieties. Hence making it feasible for industrial-scale biosynthesis of glucoside derivatives by bypassing the demand and use of the more expensive UDP-glucose cofactor that is used in most work that involves glucosylation.
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Biosynthesis of Apigenin Glucosides in Engineered Corynebacterium glutamicum
Obed Jackson Amoah,Samir Bahadur Thapa,Su Yeong Ma,Hue Thi Nguyen,Morshed Md Zakaria,Jae Kyung Sohng The Korean Society for Microbiology and Biotechnol 2024 Journal of microbiology and biotechnology Vol.34 No.5
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 biofuels. 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℃, 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℃. 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.
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Biosynthesis of fucosyllactose in engineered Corynebacterium glutamicum
Hue Thi Nguyen,Obed Jackson Amoah,Su Yeong Ma,Jae Kyung Sohng 한국당과학회 2022 한국당과학회 학술대회 Vol.2022 No.07
Human milk oligosaccharides (HMOs) have been proven to be a key ingredient found in breast milk and beneficial to infant health. Structurally, the fucose-containing oligosaccharides are major components, with a proportion of approximately 50–80% in the HMOs. In particular, 2-fucosyllactose (2’-FL) and 3-fucosyllactose (3-FL) are 2 major components of the abundant HMO component. Importantly, 2’-FL is approved for use in infant formula for improving probiotic functions by the FDA of the USA and the European Food Safety Authority, while 3-FL can be used as a prebiotic to improve the growth of beneficial microorganisms. Over the last decade, various approaches have been conducted for fucosyllactose (FL) production, including chemical synthesis, enzymatic conversion, and microbial fermentation. Most studies were performed in Escherichia coli due to the simple culture conditions and availability of various genetic tools. Besides E. coli, Bacillus subtilis and Saccharomyces cerevisiae have also been selected as the hosts for the production of FL due to their generally regarded safe (GRAS) status. As a type of GRAS strain, Corynebacterium glutamicum has been widely used in the biosynthesis of food-grade, high-value-added products, including lysine, glutamic acid, violacein and resveratrol. In this study, the biosynthetic enzymes of FL were condon-optimized and heterologously expressed in engineered C. glutamicum for the production of FL. The trace amount production of FL was confirmed by TLC, HPLC in this study.
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