Production of chondroitin in metabolically engineered E. coli

He, Wenqin; Fu, Li; Li, Guoyun; Jones, J. Andrew; Linhardt, Robert J.; Koffas, Mattheos

doi:10.1016/j.ymben.2014.11.003

Cited by 121 publications

(98 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…A recombinant E. coli BL21 strain, which produces nearly 2 g/L of heparosan, was prepared by introducing four heparosan biosynthetic genes ( KfiA- D) from E. coli K5 [97]. The production of the structurally related chondroitin CPS from glucose using an engineered E. coli was also recently reported [98]. Control of the molecular weight of the heparosan CPS obtained from E. coli K5 may also be required [99,100].…”

Section: Bioengineering Ulmwh Lmwh and Heparinmentioning

confidence: 99%

Bioengineered heparins and heparan sulfates

Suflita

Linhardt

2016

Advanced Drug Delivery Reviews

Self Cite

102

View full text Add to dashboard Cite

Heparin and heparan sulfates are closely related linear anionic polysaccharides, called glycosaminoglycans, which exhibit a number of important biological and pharmacological activities. These polysaccharides, having complex structures and polydispersity, are biosynthesized in the Golgi of animal cells. While heparan sulfate is a widely distributed membrane and extracellular glycosaminoglycan, heparin is found primarily intracellularly in the granules of mast cells. While heparin has historically received most of the scientific attention for its anticoagulant activity, interest has steadily grown in the multi-faceted role heparan sulfate plays in normal and pathophysiology. The chemical synthesis of these glycosaminoglycans is largely precluded by their structural complexity. Today, we depend on livestock animal tissues for the isolation and the annual commercial production of hundred ton quantities of heparin used in the manufacture of anticoagulant drugs and medical device coatings. The variability of animal-sourced heparin and heparan sulfates, 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, driven by both therapeutic applications and as probes to study their natural functions. This review focuses on the complex biology of these glycosaminoglycans in human health and disease, and the use of recombinant technology in the chemoenzymatic synthesis and metabolic engineering of heparin and heparan sulfates.

show abstract

Section: Bioengineering Ulmwh Lmwh and Heparinmentioning

confidence: 99%

Bioengineered heparins and heparan sulfates

Suflita

Linhardt

2016

Advanced Drug Delivery Reviews

Self Cite

102

View full text Add to dashboard Cite

show abstract

“…Homologous overexpression of kfoC in K4 reportedly increased CPS productivity by 100%, although the compatibility of the plasmid in wild type E. coli K4 strain may cause gene expression stability issues during scale-up (Cimini et al 2010). A recent publication suggests that utilizing engineered IS2 transposable elements from K4 fused with kfoC will result in a more stable overexpression system upon integration into the genome (He et al 2015). A 2.5-fold increase is reported in large-scale fermenter (He et al 2015).…”

Section: Bioengineering Approachesmentioning

confidence: 99%

“…A recent publication suggests that utilizing engineered IS2 transposable elements from K4 fused with kfoC will result in a more stable overexpression system upon integration into the genome (He et al 2015). A 2.5-fold increase is reported in large-scale fermenter (He et al 2015). Moreover, a single mutation on chondroitin polymerase kfoC (R313Q) potentially enhances the affinity of the polymerase to the UDP-GalNAc increasing K4 CPS productivity by 80% (Zanfardino et al 2010).…”

Section: Bioengineering Approachesmentioning

confidence: 99%

“…One intermediate approach can be envisioned using currently available technology to couple metabolic engineering with biotransformation. For example, the production of chondroitin CPS from glucose using an engineered E. coli is currently possible (He et al 2015). Moreover, PAPS can be produced from ATP and inorganic sulfate using three E. coli expressed recombinant enzymes, adenosine 5’-triphosphate (ATP) sulfurylase, adenosine 5’-phosphosulfate (APS) kinase, and pyrophosphatase (Zhao et al 2011) (Figure 6B).…”

Section: Conclusion and Future Prospectsmentioning

confidence: 99%

See 1 more Smart Citation

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

Suflita

et al. 2015

Appl Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

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.

show abstract

“…K4 and K5 polysaccharides are consolidated precursors of chondroitin and heparin; however, CS is still obtained from limited and potentially harmful, animal sources such as chicken keel, shark fins, and swine or bovine trachea. Several efforts have been made to use E. coli K4, or its biosynthetic machinery in a different background, to generate cell factories for the production of chondroitin, and encouraging results were obtained (Cimini et al 2013, 2015; He et al 2015; Jin et al 2016). The reason lies in the fact that the capsular polysaccharide of this uropathogenic strain, composed of alternating residues of N -acetylgalactosamine (GalNAc) and glucuronic acid (GlcA), has a structure that closely resembles the backbone of CS.…”

Section: Introductionmentioning

confidence: 99%

Engineering S. equi subsp. zooepidemicus towards concurrent production of hyaluronic acid and chondroitin biopolymers of biomedical interest

et al. 2017

View full text Add to dashboard Cite

Glycosaminoglycans, such as hyaluronic acid and chondroitin sulphate, are not only more and more required as main ingredients in cosmeceutical and nutraceutical preparations, but also as active principles in medical devices and pharmaceutical products. However, while biotechnological production of hyaluronic acid is industrially established through fermentation of Streptococcus spp. and recently Bacillus subtilis, biotechnological chondroitin is not yet on the market. A non-hemolytic and hyaluronidase negative S. equi subsp. zooepidemicus mutant strain was engineered in this work by the addition of two E. coli K4 genes, namely kfoA and kfoC, involved in the biosynthesis of chondroitin-like polysaccharide. Chondroitin is the precursor of chondroitin sulphate, a nutraceutical present on the market as anti-arthritic drug, that is lately being studied for its intrinsic bioactivity. In small scale bioreactor batch experiments the production of about 1.46 ± 0.38 g/L hyaluronic acid and 300 ± 28 mg/L of chondroitin with an average molecular weight of 1750 and 25 kDa, respectively, was demonstrated, providing an approach to the concurrent production of both biopolymers in a single fermentation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-017-0364-7) contains supplementary material, which is available to authorized users.

show abstract

Production of chondroitin in metabolically engineered E. coli

Cited by 121 publications

References 34 publications

Bioengineered heparins and heparan sulfates

Bioengineered heparins and heparan sulfates

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

Engineering S. equi subsp. zooepidemicus towards concurrent production of hyaluronic acid and chondroitin biopolymers of biomedical interest

Contact Info

Product

Resources

About