Engineered Escherichia coli Nissle 1917 with urate oxidase and an oxygen-recycling system for hyperuricemia treatment Rui Zhao et al. Gut Microbes. 2022 Jan-Dec. Free PMC article Abstract Hyperuricemia is the second most prevalent metabolic disease to human health after diabetes. Only a few clinical drugs are available, and most of them have serious side effects. The human body does not have urate oxidase, and uric acid is secreted via the kidney or the intestine. Reduction through kidney secretion is often the cause of hyperuricemia. We hypothesized that the intestine secretion could be enhanced when a recombinant urate-degrading bacterium was introduced into the gut. We engineered an Escherichia coli Nissle 1917 strain with a plasmid containing a gene cassette that encoded two proteins PucL and PucM for urate metabolism from Bacillus subtilis, the urate importer YgfU and catalase KatG from E. coli, and the bacterial hemoglobin Vhb from Vitreoscilla sp. The recombinant E. coli strain effectively degraded uric acid under hypoxic conditions. A new method to induce hyperuricemia in mice was developed by intravenously injecting uric acid. The engineered Escherichia coli strain significantly lowered the serum uric acid when introduced into the gut or directly injected into the blood vessel. The results support the use of urate-degrading bacteria in the gut to treat hyperuricemia. Direct injecting bacteria into blood vessels to treat metabolic diseases is proof of concept, and it has been tried to treat solid tumors. Keywords: Escherichia coli nissle 1917; catalase; hemoglobin; hyperuricemia; urate oxidase; uric acid. Conflict of interest statement No potential conflict of interest was reported by the author(s). Figures Figure 1. The schematic diagram of an engineered EcN strain for hyperuricemia was engineered to degrade UA via the pathway in Bacillus subtilis. The ygfU gene was co-expressed to facilitate UA transport, VHb was used to improve oxygen utilization, and H2O2, a byproduct of UOX, was eliminated by KatG. The new method to induce hyperuricemia in mice was established by intravenously injecting high concentrated UA. The recombinant strain was used to treat the hyperuricemia mice by oral administration or intravenous injection. Both therapies decreased UA levels of the mice. Figure 2. The optimization of UA degradation by engineering EcN cells. (a-b). UA degradation by using crude enzymes (a) or whole cells (b) of engineered EcN expressing PucLT in different plasmids under the control of different promoters. (c) UA degradation by EcN whole cells with PucL, PucLT, and PucLM. (d) UA degradation by EcN whole cells by co-expressing ygfU. The degradation curves were determined in HEPES buffer (pH = at OD600 = for whole cells or with proteins at mg/mL for enzymatic assays. The UA degradation ability of these whole cells or crude enzyme were assayed at defined time intervals. Three parallel experiments were executed to obtain averages and calculate STDEV. The one-way ANOVA method was used to calculate the p value. The Q values were calculated to get the false discovery rate (FDR). Q ‘NS’ was marked; Q ‘ns’ was marked; Q .05, ‘ns’ was marked; p .05, ‘ns’ was marked; p < .05, ‘*’ was marked; p < .01, ‘**’ was marked; p < .001, ‘***’ was marked. Similar articles Management of hyperuricemia with rasburicase review. de Bont JM, Pieters R. de Bont JM, et al. Nucleosides Nucleotides Nucleic Acids. 2004 Oct;23(8-9):1431-40. doi: Nucleosides Nucleotides Nucleic Acids. 2004. PMID: 15571272 Review. Construction and expression of recombinant uricase‑expressing genetically engineered bacteria and its application in rat model of hyperuricemia. Cai L, Li Q, Deng Y, Liu X, Du W, Jiang X. Cai L, et al. Int J Mol Med. 2020 May;45(5):1488-1500. doi: Epub 2020 Feb 24. Int J Mol Med. 2020. PMID: 32323736 Free PMC article. Cloning and expression of a urate oxidase and creatinine hydrolase fusion gene in Escherichia coli. Cheng X, Liu F, Zhang Y, Jiang Y. Cheng X, et al. Ren Fail. 2013;35(2):275-8. doi: Epub 2013 Jan 9. Ren Fail. 2013. PMID: 23297748 Identification of a Formate-Dependent Uric Acid Degradation Pathway in Escherichia coli. Iwadate Y, Kato JI. Iwadate Y, et al. J Bacteriol. 2019 May 8;201(11):e00573-18. doi: Print 2019 Jun 1. J Bacteriol. 2019. PMID: 30885932 Free PMC article. Serum uric acid-lowering therapies: where are we heading in management of hyperuricemia and the potential role of uricase. Bomalaski JS, Clark MA. Bomalaski JS, et al. Curr Rheumatol Rep. 2004 Jun;6(3):240-7. doi: Curr Rheumatol Rep. 2004. PMID: 15134605 Review. Cited by Effect and Potential Mechanism of Lactobacillus plantarum Q7 on Hyperuricemia in vitro and in vivo. Cao J, Bu Y, Hao H, Liu Q, Wang T, Liu Y, Yi H. Cao J, et al. Front Nutr. 2022 Jul 6;9:954545. doi: eCollection 2022. Front Nutr. 2022. PMID: 35873427 Free PMC article. References Gustafsson D, Unwin R.. The pathophysiology of hyperuricaemia and its possible relationship to cardiovascular disease, morbidity and mortality. BMC Nephrol. 2013;14(1):164. doi: - DOI - PMC - PubMed Kang E, S-s H, Kim DK, K-h O, Joo KW, Kim YS, Lee H. Sex-specific relationship of serum uric acid with all-cause mortality in adults with normal kidney function: an observational study. J Rheumatol. 2017;44(3):380–19. doi: - DOI - PubMed Hafez RM, Abdel-Rahman TM, Naguib RM. Uric acid in plants and microorganisms: biological applications and genetics - A review. J Adv Res. 2017;8(5):475–486. doi: - DOI - PMC - PubMed Singh G, Lingala B, Mithal A. Gout and hyperuricaemia in the USA: prevalence and trends. Rheumatology. 2019;58(12):2177–2180. doi: - DOI - PubMed Shirasawa T, Ochiai H, Yoshimoto T, Nagahama S, Watanabe A, Yoshida R, Kokaze A. Cross-sectional study of associations between normal body weight with central obesity and hyperuricemia in Japan. BMC Endocr Disord. 2020;20(1):2. doi: - DOI - PMC - PubMed Publication types MeSH terms Substances Grant support This work was supported by the National High Technology Research and Development Program of China [2018YFA0901200]; the National Natural Science Foundation of China [31870085]; the National Natural Science Foundation of China [31961133015]; Qilu Youth Scholar Startup Funding of SDU. LinkOut - more resources Full Text Sources Europe PubMed Central PubMed Central Taylor & Francis Medical MedlinePlus Health Information
Escherichia coli Nissle 1917 (EcN) is an efficient probiotic strain extensively used worldwide because of its several health benefits. Adhesion to the intestinal cells is one of the prerequisites for a probiotic strain. To identify the genes essential for the adhesion of EcN on the intestinal cells, …Among these, Escherichia coli Nissle 1917 CFS significantly inhibited biofilm formation and disperse … In the quest for mitigators of bacterial virulence, cell-free supernatants (CFS) from 25 human commensal and associated bacteria were tested for activity against Pseudomonas aeruginosa.
Further investigation of E. coli Nissle 1917 showed that no drastic differences in colonization and amplification were seen when immunocompetent and immunocompromised animals were used, and we were able to show that E. coli Nissle 1917 replicates at the border of live and necrotic tumor tissue.
Escherichia coli Nissle 1917 (EcN) is a probiotic isolated in 1917 by the German physician Alfred Nissle and is currently commercialized under the product name Mutaflor ® by AdeyPharm. The anti-inflammatory properties of EcN have been evaluated in a substantial number of mice colitis models [6,7,8]. This is attributable to EcN’s ability to
The gut is a major source of ammonia production that contributes to hyperammonemia in CLD and HE and remains the primary therapeutic target for lowering hyperammonemia. As an ammonia-lowering strategy, Escherichia coli Nissle 1917 bacterium was genetically modified to consume and convert ammonia to arginine (S-ARG).
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DOI: 10.1016/j.jconrel.2017.10.041 Corpus ID: 5363475; Doxorubicin‐conjugated Escherichia coli Nissle 1917 swimmers to achieve tumor targeting and responsive drug release @article{Xie2017DoxorubicinconjugatedEC, title={Doxorubicin‐conjugated Escherichia coli Nissle 1917 swimmers to achieve tumor targeting and responsive drug release}, author={Songzhi Xie and Long Zhao and Xiaojie Song and
The probiotic Escherichia coli strain Nissle 1917 (EcN) has been used for decades in human medicine in Central Europe for the treatment and prevention of intestinal disorders and diseases. However, the molecular mechanisms underlying its beneficial effects are only partially understood. To identify …
Escherichia coli Nissle 1917 (EcN) is a well-characterized probiotic bacterium. Although genomic comparisons of EcN with the uropathogenic E. coli strain CFT073 revealed high degrees of similarity, EcN is generally considered a non-pathogenic organism. However, as recent evidence suggests that EcN i …
Certain strains of Escherichia coli are also available in some European countries, the best known example of which is E. coli strain Nissle 1917 (EcN). EcN is marketed as a probiotic drug in two galenic presentations for oral use: enteric-coated capsules and a suspension in which 1 ml contains 10 8 viable EcN cells.
This mini review of the genetics and physical properties of Escherichia coli strain Nissle 1917 discusses the numerous genetic engineering strategies employed for EcN-based vaccine development, including recombinant plasmid transfer, genetic engineering of cryptic plasmids or the EcN chromosome, EcN bacterial ghosts and its outer membrane vesicles.
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The beneficial effects of genetically engineered Escherichia coli Nissle 1917 in obese C57BL/6J mice Jie Ma 1,2 , Junrui Wang 3,4 ,LuXu 2 , Yuanqi Liu 5 and Jianwen Gu 6
Background: Pathogenic adherent-invasive Escherichia coli have been isolated from ileal lesions of Crohn's disease. Aim: : To investigate the non-pathogenic E. coli strain Nissle 1917 (Mutaflor) as possible maintenance therapy in inflammatory bowel disease by testing its ability to prevent adherent-invasive E. coli strains from adhering to and invading human intestinal epithelial cells in vitro.
Lac operon is the standard regulator used to control the orthogonality of T7RNA polymerase (T7RNAP) and T7 promoter inEscherichia coli BL21(DE3) strain for protein expression. However,E. coliNissle 1917 (EcN), the unique probiotic strain, has seldom been precisely adapted to the T7 system. Herein, we applied bioinformatics analysis on Lac operon from different strains, and it was observed that
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OBJECTIVES Different probiotic strains used in clinical trials have shown prophylactic properties in different inflammatory diseases of the gastrointestinal tract. This study was aimed to investigate the influence of Escherichia coli strain Nissle 1917 (EcN) components on the integrity of the Caco-2 cell monolayer (human adenocarcinoma cell line).
Although Escherichia coli Nissle 1917 (EcN) has been used therapeutically for over a century, the determinants of its probiotic properties remain elusive. EcN produces two siderophore-microcins (Mcc) responsible for an antagonistic activity against other Enterobacteriaceae. EcN also synthesizes the genotoxin colibactin encoded by the pks island.
Aims To assess protective efficacy of genetically modified Escherichia coli Nissle 1917 (EcN) on metabolic effects induced by chronic consumption of dietary fructose. Materials and Methods EcN was genetically modified with fructose dehydrogenase (fdh) gene for conversion of fructose to 5-keto-D-fructose and mannitol-2-dehydrogenase (mtlK) gene for conversion to mannitol, a prebiotic. Charles Despite its myriad benefits, the safety of E. coli Nissle 1917 as a probiotic has been questioned. Gronback et al showed that when both the host gut microbiota and adaptive immunity are defective in mice, E. coli Nissle 1917 was able to translocate through the epithelial layer, leading to dissemination, septicemia, and death of the animals .c0iFJ.