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

Abstract

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.

Glycosaminoglycan‐Based Biohybrid Hydrogels: A Sweet and Smart Choice for Multifunctional Biomaterials

“…[185] The metabolic engineering of Golgi-lacking cells, such as E. coli , is considered even more difficult but is being targeted by dedicated ongoing studies. For a recent review of the expression techniques currently used for the formation of GAGs, the reader is referred to Suflita et al [186] …”

Bioengineered heparins and heparan sulfates

“…The current worldwide heparin market is evenly split between the use of heparin and LMWHs, with very little synthetic ULMWH being used [59,71]. Only ~5% of the heparin market is comprised of the expensive synthetic ULMWH, fondaparinux, which is used primarily when side effects such as heparin-induced thrombocytopenia are anticipated.…”

“…Only ~5% of the heparin market is comprised of the expensive synthetic ULMWH, fondaparinux, which is used primarily when side effects such as heparin-induced thrombocytopenia are anticipated. Demand for fondaparinux remains low due to its high cost, poorer pharmacological profile, irreversibility and ineffectiveness in a number of applications [59,71]. A new synthetic ULMWH called semuloparin, which is prepared by a selective and controlled depolymerization of heparin through a β-elimination reaction, was recently developed for the prevention of venous thromboembolism [72].…”

Glycolysis and Its Metabolic Engineering Applications

“…[27][28][29] Instead of extraction from animal tissues, animal polysaccharides like chondroitin and heparosan have also been engineered in microbial hosts. 30 The biosynthetic pathway of heparosan, the precursor of heparin, has been engineered in non pathogenic E. coli BL21 via expression of kfiABCD operon and heparosan was produced with a titer of 1.88 g/L in 3L bioreactor. 31 The chondroitin pathway consisting of three genes kfoA (encoding uridine diphosphate (UDP)GlcNAc 4epimerase), kfoC (chondroitin polymerase), and kfoF (UDPglucose dehydrogenase) from E. coli K4 was introduced into the nonpathogenic E. coli BL21Star TM (DE3) and 2.4 g/L chondroitin was produced in 2 L bioreactor.…”