High density fermentation of probiotic E. coli Nissle 1917 towards heparosan production, characterization, and modification

Datta, Payel; Fu, Li; Brodfuerer, Paul; Dordick, Jonathan S.; Linhardt, Robert J.

doi:10.1007/s00253-020-11079-9

Cited by 19 publications

(7 citation statements)

References 48 publications

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“…For this reason, the insertion of Bt. plebeius β-agarase genes into S. boulardii was used [203]. Other probiotics were modified to biosynthesize valuable polysaccharides, such as heparosana precursor to heparin that can be used to deliver certain drugs.…”

Section: Recombinant Probiotics As "Cell Factories" or For The Synthe...mentioning

confidence: 99%

Recombinant Probiotic Preparations: Current State, Development and Application Prospects

Khablenko

Danylenko

Yalovenko

et al. 2023

Innov Biosyst Bioeng

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The article is devoted to the latest achievements in the field of research, development, and implementation of various types of medicinal products based on recombinant probiotics. The benefits of probiotics, their modern use in medicine along with the most frequently used genera and species of probiotic microorganisms were highlighted. The medicinal and therapeutic activities of the studied probiotics were indicated. The review suggests various methods of creating recombinant probiotic microorganisms, including standard genetic engineering methods, as well as systems biology approaches and new methods of using the CRISPR-Cas system. The range of potential therapeutic applications of drugs based on recombinant probiotics was proposed. Special attention was paid to modern research on the creation of new, more effective recombinant probiotics that can be used for various therapeutic purposes. Considering the vast diversity of therapeutic applications of recombinant probiotics and ambiguous functions, their use for the potential treatment of various common human diseases (non-infectious and infectious diseases of the gastrointestinal tract, metabolic disorders, and allergic conditions) was investigated. The prospects for creating different types of vaccines based on recombinant probiotics together with the prospects for their implementation into medicine were considered. The possibilities of using recombinant probiotics in veterinary medicine, particularly for the prevention of domestic animal diseases, were reviewed. The prospects for the implementation of recombinant probiotics as vaccines and diagnostic tools for testing certain diseases as well as modeling the work of the human digestive system were highlighted. The risks of creation, application, including the issues related to the regulatory sphere regarding the use of new recombinant microorganisms, which can potentially enter the environment and cause unforeseen circumstances, were outlined.

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Section: Recombinant Probiotics As "Cell Factories" or For The Synthe...mentioning

confidence: 99%

Recombinant Probiotic Preparations: Current State, Development and Application Prospects

Khablenko

Danylenko

Yalovenko

et al. 2023

Innov Biosyst Bioeng

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show abstract

“…Because the availability of heparosan was widely concerned, various efforts have been made to increase the production of heparosan (Datta et al, 2021;Z. Wang et al, 2010Yan et al, 2015;C.…”

Section: Introductionmentioning

confidence: 99%

“…Because the availability of heparosan was widely concerned, various efforts have been made to increase the production of heparosan (Datta et al, 2021; Z. Wang et al, 2010, 2011; Yan et al, 2015; C. Zhang et al, 2012). Heparosan is synthesized and exported as extracellular capsular polysaccharide by Escherichia coli K5, Pasteurella multicida Type D, and E. coli strain Nissle 1917 (EcN) (Cress, Linhardt, et al, 2013; Cress, Greene, et al 2013; De Angelis, 2012; De Angelis & White, 2002; Zhu et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Chromosome evolution of Escherichia coli Nissle 1917 for high‐level production of heparosan

Gong

Wang

et al. 2023

Biotech & Bioengineering

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Heparosan is a crucial-polysaccharide precursor for the chemoenzymatic synthesis of heparin, a widely used anticoagulant drug. Presently, heparosan is mainly extracted with the potential risk of contamination from Escherichia coli strain K5, a pathogenic bacterium causing urinary tract infection. Here, a nonpathogenic probiotic, E. coli strain Nissle 1917 (EcN), was metabolically engineered to carry multiple copies of the 19-kb kps locus and produce heparosan to 9.1 g/L in fed-batch fermentation. Chromosome evolution driven by antibiotics was employed to amplify the kps locus, which governed the synthesis and export of heparosan from EcN at 21 mg L −1 OD −1 . The average copy number of kps locus increased from 1 to 24 copies per cell, which produced up to 104 mg L -1 OD −1 of heparosan in the shaking flask cultures of engineered strains. The following in-frame deletion of recA stabilized the recombinant duplicates of chromosomal kps locus and the productivity of heparosan in continuous culture for at least 56 generations. Fed-batch fermentation of the engineered strain EcN8 was carried out to bring the yield of heparosan up to 9.1 g/L.Heparosan from the fermentation culture was further purified at a 75% overall recovery. The structure of purified heparosan was characterized and further modified by N-sulfotransferase with 3′-phosphoadenosine-5′-phosphosulfate as the sulfodonor. The analysis of element composition showed that heparosan was N-sulfated by over 80%. These results indicated that duplicating large DNA cassettes up to 19-kb, followed by high-cell-density fermentation, was promising in the large-scale preparation of chemicals and could be adapted to engineer other industrial-interest bacteria metabolically.

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“…In addition, the high risk of viral contamination to animals results in the difficulty in quality control 21 . In the last decades, microbial fermentation was developed for producing GAGs and other polysaccharides in large scale 22–26 . Especially, HA as an important non‐sulfated GAG has been produced from the cell cultures of several strains, which has been realized the industrialization presently 26 .…”

Section: Introductionmentioning

confidence: 99%

“…21 In the last decades, microbial fermentation was developed for producing GAGs and other polysaccharides in large scale. [22][23][24][25][26] Especially, HA as an important non-sulfated GAG has been produced from the cell cultures of several strains, which has been realized the industrialization presently. 26 Researchers also focused on the fermentation of CS in bacteria since its mature expression system and manipuility in metabolic engineering.…”

Section: Introductionmentioning

confidence: 99%

Conjugation of a phenanthrene‐imidazole fluorophore with the chondroitin sulfate generated from Escherichia coliK4 polysaccharide

Huang

Wang

et al. 2021

J of Applied Polymer Sci

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Escherichia coli K4 expressed a fructosylated chondroitin (K4), which had a chondroitin structure after the defructosylation treatment. Polysaccharide K4 as a preferred precursor was modified to produce chondroitin sulfate A (CSA). Here, glycosaminoglycans including K4, defructosylated K4, and CSA were conjugated with fluorophore A, which were optimized under three reaction conditions to give the maximum DS of 2.9, 6.5, and 8.5%, respectively. All of conjugates displayed luminescence response linearly to concentration. And they exhibited good storage stability in luminescence and dispersion within 24 h. The conjugate CSA‐A with substituted degree of 6.4% was chosen for the bioimaging study in vitro. CSA‐A showed nontoxic to astrocytes with concentrations lower than 100 μg/ml. It was internalized effectively by astrocytes after incubation for 24 h.

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High density fermentation of probiotic E. coli Nissle 1917 towards heparosan production, characterization, and modification

Cited by 19 publications

References 48 publications

Recombinant Probiotic Preparations: Current State, Development and Application Prospects

Recombinant Probiotic Preparations: Current State, Development and Application Prospects

Chromosome evolution of Escherichia coli Nissle 1917 for high‐level production of heparosan

Conjugation of a phenanthrene‐imidazole fluorophore with the chondroitin sulfate generated from Escherichia coliK4 polysaccharide

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