Abstract
Glycosyltransferase from Bacillus licheniformis DSM13 (YjiC) was used for enzymatic modification of emodin and aloe-emodin in vitro and in vivo. In order to increase the availability of UDP-glucose, three genes involved in the production of precursors of NDP-sugar in Escherichia coli BL21 (DE3) viz. d-glucose phosphate isomerase (pgi), d-glucose-6-phosphate dehydrogenase (zwf), and UDP-sugar hydrolase (ushA) were deleted and glucose-1-phosphate urididyltransferase (galU) gene was over expressed. To improve the yield of the products; substrate, time and media parameters were optimized, and the production was scaled up using a 3 L fermentor. The maximum yield of glycosylated products of emodin (emodin-O-β-d-glucoside) and aloe-emodin (aloe-emodin-O-β-d-glucoside) were approximately 144 µM (38 mg/L) and 168 µM (45 mg/L) respectively, representing almost 72 % and 84 % bioconversion of emodin and aloe-emodin when 200 µM of emodin and aloe-emodin were supplemented in the culture. Additionally, the emodin and aloe emodin major glycosylated products exhibited the highest stability at pH 8.0 and the stability of products was up to 70 °C and 60 °C respectively. Furthermore, the biological activities of emodin and its major glucoside (P1) were compared and their anti-cancer activities were assayed in several cancer cell lines. The results demonstrate that YjiC has the capacity to catalyze the glycosylation of these aromatic compounds and that glycosylation of anthraquinones enhances their aqueous solubility while retaining their biological activities.
Similar content being viewed by others
References
Acevedo-Duncan M, Russell C, Patel S, Patel R (2004) Aloe-emodin modulates PKC isozymes, inhibit proliferation, and induces apoptosis in U-373MG glioma cells. Int Immunopharmacol 4:1775–1784
Anand S, Muthusamy VS, Sujatha S, Sangeetha KN, Raja RB, Sudhgar S, Devi NP, Laxmi BS (2010) Aloe emodin glycosides stimulates transport and glycogen storage through PI3K dependent mechanism in L6 myotubes and inhibits adipocyte differentiation in 3T3L1 adipocytes. FEBS Lett 584:3170–3178
Astumi S, Cann AF, Connor MR, Shen CR, Smith KM, Beynildsen MP et al (2008) Metabolic engineering of Escherichia coli for 1-butanol production. Metab Eng 10:305–311
Chang CH, Lin CC, Yang JJ, Namba T, Hattori M (1996) Anti-inflammatory effects of emodin from Ventilago leiocarpa. Am J Chin Med 24:139–142
Dave H, Ledwani L (2012) A review on anthraquinones isolated from Cassia species and their applications. Indian J Nat Prod Res 3:291–319
Du J, Shao Z, Zhao H (2011) Engineering microbial factories for synthesis of value-added products. J Ind Microbiol Biotechnol 38:873–890
Eshun K, He Q (2004) Aloe vera: a valuable ingredient for the food, pharmaceutical and cosmetic industries- a review. Crit Rev Food Sci Nutr 44:91–96
Gurung RB, Kim EH, Oh TJ, Sohng JK (2013) Enzymatic synthesis of apigenin glucosides by glucosyltransferase (YjiC) from Bacillus licheniformis DSM 13. Mol Cells 36:355–361
Huang Q, Lu G, Shen HM, Chung MC, Ong CN (2007) Anti-cancer properties of anthraquinones from rhubarb. Med Res Rev 27:609–630
Koirala N, Pandey RP, Parajuli P, Jung HJ, Sohng JK (2014a) Methylation and subsequent glycosylation of 7,8-dihydroxyflavone. J Biotechnol 184:128–137
Koirala N, Pandey RP, Thang DV, Jung HJ, Sohng JK (2014b) Glycosylation and subsequent malonylation of isoflavonoids in E. coli: strain development, production and insights into future metabolic perspectives. J Ind Microbiol Biotechnol 41:1647–1658
Kuo PL, Lin TC, Lin CC (2002) The antiproliferative activity of aloe-emodin is through p53-dependent and p21-dependent apoptotic pathway in human hepatoma cell lines. Life Sci 71:1879–1892
Lee HZ, Hsu SL, Liu MC, Wu CH (2001) Effects and mechanisms of aloe-emodin on cell death in human lung squamous cell carcinoma. Eur J Pharmacol 431:287–295
Leonard E, Yan Y, Fowler ZL, Li Z, Lim CG, Lim KH, Koffas MAG (2008) Strain improvement of recombinant Escherichia coli for efficient production of plant flavonoids. Mol Pharm 5:257–265
Liu T, Khosla C (2010) Genetic engineering of Escherichia coli for biofuel production. Annu Rev Genet 44:53–69
Pandey RP, Malla S, Simkhada D, Kim BG, Sohng JK (2013a) Production of 3-O-xylosyl quercetin in Escherichia coli. Appl Microbiol Biotechnol 97:1889–1901
Pandey RP, Parajuli P, Koirala N, Sohng JK (2013b) Probing of 3-hydroxyflavone for in vitro glycorandamization of flavonols by YjiC. Appl Environ Microbiol 79:6833–6838
Pandey RP, Gurung RB, Parajuli P, Koirala N, Tuoi le T, Sohng JK (2014) Assessing acceptor substrate promiscuity of YjiC-mediated glycosylation towards flavonoids. Carbohydr Res 393:26–31
Pecere T, Gazzola MV, Mucignat C, Parolin C, Vecchia FD, Cavaggioni A, Basso G, Diaspro A, Salvato B, Carli M, Palu G (2000) Aloe-emodin is a new type of anticancer agent with selective activity against neuroectodermal tumors. J Cancer 60:2800–2804
Reynolds T (1985) The compounds in aloe leaf exudates: a review. Bot J Linn Soc 90:157–177
Rodriguez-Diaz J, Yebra MJ (2011) Enhanced UDP-glucose and UDP-galactose by homologous overexpression of UDP-glucose pyrophosphorylase in Lactobacillus casei. J Biotechnol 154:212–215
Ruffing A, Chen RR (2006) Metabolic engineering of microbes for oligosaccharide and polysaccharide synthesis. Microbiol Cell Fact 5:25
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn edn. Cold Spring Harbor, New York
Thorson JS, Barton WA, Hoffmeister D, Albermann C, Nikolov DB (2004) Structure-based enzyme engineering and its impact on in vitro glycorandomization. ChemBioChem 5:16–25
Vogt T, Jones P (2000) Glycosyltransferases in plant natural product synthesis: characterization of a supergene family. Trends Plant Sci 5:380–386
Wang HH, Chung JG (1997) Emodin-induced inhibition of growth and DNA damage in the Helicobacter pylori. Curr Microbiol 35:262–266
Wang C, Zhang D, Ma H, Liu J (2007) Neuroprotective effects of emodin-8-O-β-D-glucoside in vivo and in vitro. Eur J Pharmacol 577:58–63
Wang W, Zhou Q, Liu L, Zou K (2012) Anti-allergic activity of emodin on IgE-mediated activation in RBL-2H3 cells. Pharmacol Rep 64:1216–1222
Williams GJ, Yang J, Zhang C, Thorson JS (2011) Recombinant E. coli prototype strains for in vivo glycorandomization. ACS Chem Biol 6:95–100
Xue J, Ding W, Liu Y (2010) Anti-diabetic effects of emodin involved in the activation of PPARγ on high-fat diet-fed and low dose of streptozotocin-indiced diabetic mice. Fitoterapia 81:173–177
Acknowledgments
This work was carried out with the support of “Cooperative Research Program for Agriculture Science and Technology Development (Project title: National Agricultural Genome Program, No. PJ01045705)” Rural Development Administration, Republic of Korea.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ghimire, G.P., Koirala, N., Pandey, R.P. et al. Modification of emodin and aloe-emodin by glycosylation in engineered Escherihia coli . World J Microbiol Biotechnol 31, 611–619 (2015). https://doi.org/10.1007/s11274-015-1815-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11274-015-1815-4