Production and purification of an extracellularly produced K4 polysaccharide from Escherichia coli

Manzoni, M.; Bergomi, Silvia; Molinari, Francesco; Cavazzoni, V.

doi:10.1007/bf00143456

Cited by 24 publications

(18 citation statements)

References 5 publications

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“…Several studies have been performed to improve the production and yield of K4 CPS comprised of fructosylated chondroitin (Manzoni et al 1996; Zoppetti et al 2004; Cimini et al 2010; Restaino et al 2011). The growth of E. coli K4 has been optimized by altering medium composition, including the use of glucose, glycerol and soya peptone.…”

Section: Bioengineering Approachesmentioning

confidence: 99%

Heparin and related polysaccharides: synthesis using recombinant enzymes and metabolic engineering

Suflita

et al. 2015

Appl Microbiol Biotechnol

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Glycosaminoglycans are linear anionic polysaccharides that exhibit a number of important biological and pharmacological activities. The two most prominent members of this class of polysaccharides are heparin/heparan sulfate and the chondroitin sulfates (including dermatan sulfate). These polysaccharides, having complex structures and polydispersity, are biosynthesized in the Golgi of most animal cells. The chemical synthesis of these glycosaminoglycans is precluded by their structural complexity. Today, we depend on food animal tissues for their isolation and commercial production. Ton quantities of these glycosaminoglycans are used annually as pharmaceuticals and nutraceuticals. The variability of animal-sourced glycosaminoglycans, their inherent impurities, the limited availability of source tissues, the poor control of these source materials, and their manufacturing processes, suggest a need for new approaches for their production. Over the past decade there have been major efforts in the biotechnological production of these glycosaminoglycans. This mini-review focuses on the use of recombinant enzymes and metabolic engineering for the production of heparin and chondroitin sulfates.

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Section: Bioengineering Approachesmentioning

confidence: 99%

Heparin and related polysaccharides: synthesis using recombinant enzymes and metabolic engineering

Suflita

et al. 2015

Appl Microbiol Biotechnol

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“…COG classification of the 220 significant regulated proteins is shown in Figure 6. The 77 up-regulated proteins were mainly distributed in amino acid transport and metabolism (12), cell wall/membrane/envelope biogenesis (11), signal transduction mechanisms (9), and the 143 down-regulated proteins were mainly in Amino acid transport and metabolism (38), energy production and conversion (26), carbohydrate transport and metabolism (23). The accession number, protein name, peptides and ratio Table S4.…”

Section: Effects Of Slya Overexpression On K4cps Production In Shake mentioning

confidence: 99%

“…Since then, scientists have been investigating the production of CS by fermentation with E . coli K4 . To date, various techniques have been proposed to improve the production of K4CPS, including optimizing the fermentation conditions and genetic engineering.…”

Section: Introductionmentioning

confidence: 99%

“…8 Since then, scientists have been investigating the production of CS by fermentation with E. coli K4. 9 To date, various techniques have been proposed to improve the production of K4CPS, including optimizing the fermentation conditions and genetic engineering. By using E. coli K4, a yield of 1.4 g/L K4CPS and a cell density of 56 g CWW /L were obtained by optimizing the composition of the medium in fed-batch cultivations.…”

Section: Introductionmentioning

confidence: 99%

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Transcriptional engineering of Escherichia coli K4 for fructosylated chondroitin production

Yang

Zou

et al. 2013

Biotechnology Progress

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The capsule polysaccharide (CPS) of Escherichia coli K4 (K4CPS) is identical to fructosylated chondroitin, which can be modified to chondroitin sulfate, a commercially valuable biopolymer commonly used in pharmaceutical applications. In this study, we homologously overexpressed the transcriptional regulator SlyA to enhance the expression of K4 capsule gene cluster and production of CPS. The iTRAQ quantificaton of proteomics revealed 77 up-regulated proteins and 143 down-regulated proteins in E. coli THslyA. Most enzymes of glycolysis and citrate cycle pathway were weakened, while proteins associated with K4CPS synthesis were up-regulated, showing a shift of carbon flux from cell growth to K4CPS production. Further, the production of K4CPS by the recombinant strain was 1 and 2.6 g/L in a shake flask and 7-L batch bioreactor, which was 1.85- and 1.53-fold higher than that of the wild-type strain, respectively. Thus, this study provides a viable strategy for improving the production of K4CPS through a transcriptional-level manipulation.

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“…Manzoni and co‐workers were the first to describe a biotechnological process exploiting E. coli O5:K4:H4 for the production of K4 polysaccharide [41]. The growth medium described by Rodriguez et al [57] was used to set up a 14 L batch fermentation.…”

Section: Cs and Chondroitin Productionmentioning

confidence: 99%

Production of glucuronic acid‐based polysaccharides by microbial fermentation for biomedical applications

Cimini

Rosa

Schiraldi

2011

Biotechnology Journal

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This review provides an overview of the properties, different biosynthetic machineries, and biotechnological production processes of four microbially derived glucuronic acid-based polysaccharides that are of interest for diverse biomedical purposes. In particular, the utilization of hyaluronic acid and heparin sulfate in high-value medical applications is already well established, whereas chondroitin sulfate and alginate show high potential within this ever-growing field. Furthermore, new strategies exploiting genetically engineered microorganisms generated through improving naturally existing pathways or de novo designed ones are described. These new developments result in increased fermentation titers, and thereby, pave the way towards feasible, or at least improved, process economy. Moreover, these strategies also allow for the future possibility of producing tailor-made biopolymers with specified characteristics, even novel molecules.

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Production and purification of an extracellularly produced K4 polysaccharide from Escherichia coli

Cited by 24 publications

References 5 publications

Heparin and related polysaccharides: synthesis using recombinant enzymes and metabolic engineering

Heparin and related polysaccharides: synthesis using recombinant enzymes and metabolic engineering

Transcriptional engineering of Escherichia coli K4 for fructosylated chondroitin production

Production of glucuronic acid‐based polysaccharides by microbial fermentation for biomedical applications

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