Biosynthesis of Apigenin Glucosides in Engineered Corynebacterium glutamicum
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
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 25o 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 37o 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.
Abstract
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 25o 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 37o 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.
참고문헌 (55)
- Pandey RP, "Microbial production of natural and non-natural flavonoids : pathway engineering, directed evolution and systems", synthetic biology, 2016, pp.34, 634~662
- Corrêa MJC, "Biotransformation of chalcones by the endophytic fungus Aspergillus flavus isolated from Paspalum maritimum Trin", J. Braz. Chem. Soc, 22, pp.1333~1338 (2011)
- Tohge T, "Current understanding of the pathways of flavonoid biosynthesis in model and crop plants", J. Exp. Bot, 68, pp.4013~4028 (2017)
- Sheng H, "Metabolic engineering of microorganisms for the production of flavonoids", Front. Bioeng. Biotechnol, 8, pp.589~069 (2020)
- Panche AN, "Flavonoids : an overview", J. Nutr. Sci, 5, pp.1~15 (2016)
- Shrestha A, "Biosynthesis of flavone C-glucosides in engineered Escherichia coli", Appl. Microbiol. Biotechnol, 102, pp.1251~1267 (2018)
- Naz S, "Chrysin : pharmacological and therapeutic properties", Life Sci, 235, pp.1167~1197 (2019)
- Łużny M, "Biotransformation of methoxyflavones by selected entomopathogenic filamentous fungi", Int. J. Mol. Sci, 21, pp.1~22 (2020)
- Huang YS, "Polymethoxy flavones are responsible for the anti-inflammatory activity of citrus fruit peel", Food Chem, 119, pp.868~873 (2010)
- Wang T yang, "Bioactive flavonoids in medicinal plants : structure, activity and biological fate", Asian J. Pharm. Sci, 13, pp.12~23 (2018)
- Costa C, "Current evidence on the effect of dietary polyphenols intake on chronic diseases", Food Chem. Toxicol, 110, pp.286~299 (2017)
- Dymarska M, "Biotransformations of flavones and an isoflavone(daidzein)in cultures of entomopathogenic filamentous fungi", Molecules, 23, pp.1356~ (2018)
- Choi SH, "Glycosylation of various flavonoids by recombinant oleandomycin glycosyltransferase from Streptomyces antibioticus in batch and repeated batch modes", Biotechnol. Lett, 34, pp.499~505 (2012)
- Kawabata Y, "Formulation design for poorly water-soluble drugs based on biopharmaceutics classification system : basic approaches and practical applications", Int. J. Pharm, 420, pp.1~10 (2011)
- Jang SW, "Enzymatic synthesis of α-flavone glucoside via regioselective transglucosylation by amylosucrase from Deinococcus geothermalis", PLoS One, 13, pp.0207466~ (2018)
- Thuan NH, "Recent biotechnological progress in enzymatic synthesis of glycosides", J. Ind. Microbiol. Biotechnol, 40, pp.1329~1356 (2013)
- Xiao J, "Advances in the biotechnological glycosylation of valuable flavonoids", Biotechnol. Adv, 32, pp.1145~1156 (2014)
- Bosch D, "N-Glycosylation of plant-produced recombinant proteins", Curr. Pharm. Des, 19, pp.5503~5512 (2013)
- Thuan NH, "Engineering co-culture system for production of apigetrin in Escherichia coli", J. Ind. Microbiol. Biotechnol, 45, pp.175~185 (2018)
- Gutmann A, "Towards the synthesis of glycosylated dihydrochalcone natural products using glycosyltransferase-catalysed cascade reactions", Green Chem, 16, pp.4417~4425 (2014)
- Ji Y, "Advances on the in vivo and in vitro glycosylations of flavonoids", Appl. Microbiol. Biotechnol, 104, pp.6587~6600 (2020)
- Pandey RP, "Production of 3-O-xylosyl quercetin in Escherichia coli", Appl. Microbiol. Biotechnol, 97, pp.1889~1901 (2013)
- Wang H, "Engineering Saccharomyces cerevisiae with the deletion of endogenous glucosidases for the production of flavonoid glucosides", Microb. Cell. Fact, 15, pp.134~ (2016)
- Henke NA, "Carotenoid production by Corynebacterium: The Workhorse of Industrial Amino Acid Production as Host for Production of a Broad Spectrum of C40 and C50 Carotenoids, Carotenoids" (2017), InTech
- Kranz A, "A manually curated compendium of expression profiles for the microbial cell factory Corynebacterium glutamicum", Sci. Data, 9, pp.594~ (2022)
- Baritugo KA, "Metabolic engineering of Corynebacterium glutamicum for fermentative production of chemicals in biorefinery", Appl. Microbiol. Biotechnol, 102, pp.3915~3937 (2018)
- Wang Y, "Eliminating the capsule-like layer to promote glucose uptake for hyaluronan production by engineered Corynebacterium glutamicum", Nat. Commun, 11, pp.3120~ (2020)
- Liu W, "Efficient production of glutathione with multi-pathway engineering in Corynebacterium glutamicum", J. Ind. Microbiol. Biotechnol, 46, pp.1685~1695 (2019)
- Wieschalka S, "Bio-based production of organic acids with Corynebacterium glutamicum", Microb. Biotechnol, 6, pp.87~102 (2013)
- Kogure T, "Recent advances in metabolic engineering of Corynebacterium glutamicum for bioproduction of valueadded aromatic chemicals and natural products", Appl. Microbiol. Biotechnol, 102, pp.8685~8705 (2018)
- Jojima T, "Metabolic engineering for improved production of ethanol by Corynebacterium glutamicum", Appl. Microbiol. Biotechnol, 99, pp.1165~1172 (2015)
- Kallscheuer N, "Construction of a Corynebacterium glutamicum platform strain for the production of stilbenes and(2S)-flavanones", Metab. Eng, 38, pp.47~55 (2016)
- Milke L, "Production of plant-derived polyphenols in microorganisms : current state and perspectives", Appl. Microbiol. Biotechnol, 102, pp.1575~1585 (2018)
- Gauttam R, "Metabolic engineering of Corynebacterium glutamicum for production of UDP-N-Acetylglucosamine", Front. Bioeng. Biotechnol, 9, pp.748510~ (2021)
- Becker J, "Metabolically engineered Corynebacterium glutamicum for bio-based production of chemicals, fuels, materials, and healthcare products", Metab. Eng, 50, pp.122~141 (2018)
- Mimitsuka T, "Metabolic engineering of Corynebacterium glutamicum for cadaverine fermentation", Biosci. Biotechnol. Biochem, 71, pp.2130~2135 (2007)
- Heider SAE, "Production and glucosylation of C50 and C40carotenoids by metabolically engineered Corynebacterium glutamicum", Appl. Microbiol. Biotechnol, 98, pp.1223~1235 (2014)
- Amoah OJ, "N-Glucosylation in Corynebacterium glutamicum with YdhE from Bacillus lichenformis", Molecules, 27, pp.3405~ (2022)
- Jiang Y, "CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum", Nat. Commun, 8, pp.151~179 (2017)
- Seibold GM, "Inactivation of the phosphoglucomutase gene pgm in Corynebacterium glutamicum affects cell shape and glycogen metabolism", Biosci. Rep, 33, pp.645~654 (2013)
- Zha J, "Metabolic engineering of Corynebacterium glutamicum for anthocyanin production", Microb. Cell. Fact, 17, pp.143~ (2018)
- Mizuno H, "Elevated, non-proliferative temperatures change the profile of fermentation products in Corynebacterium glutamicum", Appl. Microbiol. Biotechnol, 105, pp.367~377 (2021)
- Wang J, "Total synthesis of apigenin", J. Chem. Res, 36, pp.121~122 (2012)
- Schmölzer K, "Sucrose synthase : a unique glycosyltransferase for biocatalytic glycosylation process development", Biotechnol. Adv, 34, pp.88~111 (2016)
- Darsandhari S, "One-Pot Multienzyme Cofactors Recycling(OPME-CR)system for lactose and non-natural saccharide conjugated polyphenol production", J. Agric. Food Chem, 66, pp.7965~7974 (2018)
- Yuan S, "Isolation and characterization of a multifunctional flavonoid glycosyltransferase from Ornithogalum caudatum with glycosidase activity", Sci. Rep, 8, pp.5886~ (2018)
- Gurung RB, "Enzymatic synthesis of apigenin glucosides by glucosyltransferase(YjiC)from Bacillus licheniformis DSM 13", Mol. Cells, 36, pp.355~361 (2013)
- Pandey RP, "Glucosylation of isoflavonoids in engineered Escherichia coli", Mol. Cells, 37, pp.172~177 (2014)
- Ververidis F, "Biotechnology of flavonoids and other phenylpropanoid-derived natural products. Part I: chemical diversity, impacts on plant biology and human health", Biotechnol. J, 2, pp.1214~1234 (2007)
- Tartik M, "Optimizing yeast for high-level production of kaempferol and quercetin", Microb. Cell. Fact, 22, pp.74~ (2023)
- Mizuno H, "Elevated, non-proliferative temperatures change the profile of fermentation products in Corynebacterium glutamicum", Appl. Microbiol. Biotechnol, 105, pp.367~377 (2021)
- Delaunay S, "An improved temperature-triggered process for glutamate production with Corynebacterium glutamicum", Enzyme Microb. Technol, 25, pp.8~9 (1999)
- Willits MG, "Bio-fermentation of modified flavonoids : an example of in vivo diversification of secondary metabolites", Phytochemistry, 65, pp.31~41 (2004)
- Wang M, "A review on flavonoid apigenin : dietary intake, ADME, antimicrobial effects, and interactions with human gut microbiota", Biomed Res. Int, 2019, pp.7010467~ (2019)
- Zhao L, "Efficient synthesis and in vitro hypoglycemic activity of rare apigenin glycosylation derivatives", Molecules, 28, pp.533~ (2023)