The functional molecular mass of the Pasteurella hyaluronan synthase is a monomer

Pummill, Philip E.; Kane, Tasha A.; Kempner, Ellis S.; DeAngelis, Paul L.

doi:10.1016/j.bbagen.2006.09.020

Cited by 4 publications

(2 citation statements)

References 29 publications

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“…An Escherichia coli -derived recombinant truncated (residues 1-703) version of PmHAS was used to make HA products (Pummill et al 2007). Three different E. coli -derived recombinant maltose-binding protein fusions with the P. multocida HEP synthase enzymes were used for making HEP: HEP synthase 1 (PmHS1), HEP synthase 2 (PmHS2) and a chimeric protein of these two enzymes called HEP synthase G (PmHS-G) (Sismey-Ragatz et al 2007; Otto et al 2012).…”

Section: Methodsmentioning

confidence: 99%

Expanding glycosaminoglycan chemical space: towards the creation of sulfated analogs, novel polymers and chimeric constructs

et al. 2017

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Glycosaminoglycans (GAGs) have therapeutic potential in areas ranging from angiogenesis, inflammation, hemostasis and cancer. GAG bioactivity is conferred by intrinsic structural features, such as disaccharide composition, glycosidic linkages and sulfation pattern. Unfortunately, the in vitro enzymatic synthesis of defined GAGs is quite restricted by a limited understanding of current GAG synthases and modifying enzymes. Our work provides insights into GAG-active enzymes through the creation of sulfated oligosaccharides, a new polysaccharide and chimeric polymers. We show that a C6-sulfonated uridine diphospho (UDP)-glucose (Glc) derivative, sulfoquinovose, can be used as an uronic acid donor, but not as a hexosamine donor, to cap hyaluronan (HA) chains by the HA synthase from the microbe Pasteurella multocida. However, the two heparosan (HEP) synthases from the same species, PmHS1 and PmHS2, could not employ the UDP-sulfoquinovose under similar conditions. Serendipitously, we found that PmHS2 co-polymerized Glc with glucuronic acid (GlcA), creating a novel HEP-like polymer we named hepbiuronic acid [-4-GlcAβ1-4-Glcα1-]n. In addition, we created chimeric block polymers composed of both HA and HEP segments; in these reactions GlcA-, but not N-acetylglucosamine-(GlcNAc), terminated GAG acceptors were recognized by their noncognate synthase for further extension, likely due to the common β-linkage connecting GlcA to GlcNAc in both of these GAGs. Overall, these GAG constructs provide new tools for studying biology and offer potential for future sugar-based therapeutics.

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Section: Methodsmentioning

confidence: 99%

Expanding glycosaminoglycan chemical space: towards the creation of sulfated analogs, novel polymers and chimeric constructs

et al. 2017

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“…It is therefore called a Class II HA synthase [17]. So far the P. multocida HA synthase is the only known member of the Class II HA synthases [49,51,52].…”

Section: Biosynthetic Pathways For Hyaluronan In Living Organismsmentioning

confidence: 99%

Production Methods for Hyaluronan

Boeriu

Springer

Kooy

et al. 2013

International Journal of Carbohydrate Chemistry

158

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Hyaluronan is a polysaccharide with multiple functions in the human body being involved in creating flexible and protective layers in tissues and in many signalling pathways during embryonic development, wound healing, inflammation, and cancer. Hyaluronan is an important component of active pharmaceutical ingredients for treatment of, for example, arthritis and osteoarthritis, and its commercial value far exceeds that of other microbial extracellular polysaccharides. Traditionally hyaluronan is extracted from animal waste which is a well-established process now. However, biotechnological synthesis of biopolymers provides a wealth of new possibilities. Therefore, genetic/metabolic engineering has been applied in the area of tailor-made hyaluronan synthesis. Another approach is the controlled artificial (in vitro) synthesis of hyaluronan by enzymes. Advantage of using microbial and enzymatic synthesis for hyaluronan production is the simpler downstream processing and a reduced risk of viral contamination. In this paper an overview of the different methods used to produce hyaluronan is presented. Emphasis is on the advancements made in the field of the synthesis of bioengineered hyaluronan.

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Chemoenzymatic synthesis with the Pasteurella hyaluronan synthase; production of a multitude of defined authentic, derivatized, and analog polymers

DeAngelis

2024

Proteoglycan Research

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Hyaluronan (HA; [‐3‐GlcNAc‐1‐beta‐4‐GlcA‐1‐beta]n), an essential matrix polysaccharide of vertebrates and the molecular camouflage coating in certain pathogens, is polymerized by “HA synthase” (HAS) enzymes. Three HAS classes have been identified with biotechnological utility, but only the Class II PmHAS from Pasteurella multocida Type A has been useful for preparation of very defined HA polymers in vitro. Two general chemoenzymatic strategies with different size products are possible: (1) repetitive step‐wise extension reactions by sequential addition of a single monosaccharide from a donor UDP‐sugar onto an acceptor (or “primer”) comprised of a short glycosaminoglycan chain (e.g., HA di‐, tri‐ or tetrasaccharide) or an artificial glucuronide yielding homogeneous oligosaccharides in the range of 2 to ~20 monosaccharide units (n = 1 to ~10), or (2) “one‐pot” polymerization reactions employing acceptor‐mediated synchronization with stoichiometric size control yielding quasi‐monodisperse (i.e., polydispersity approaching 1; very narrow size distributions) polysaccharides in the range of ~7 kDa to ~2 MDa (n = ~17 to 5000). In either strategy, acceptors containing non‐carbohydrate functionalities (e.g., biotin, fluorophores, amines) can add useful moieties to the reducing termini of HA chains at 100% efficiency. As a further structural diversification, PmHAS can utilize a variety of unnatural UDP‐sugar analogs thus adding novel groups (e.g., trifluoroacetyl, alkyne, azide, sulfhydryl) along the HA backbone and/or at its nonreducing terminus. This review discusses the current understanding and recent advances in HA chemoenzymatic synthesis methods using PmHAS. This powerful toolbox has potential for creation of a multitude of HA‐based probes, therapeutics, drug conjugates, coatings, and biomaterials.

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The functional molecular mass of the Pasteurella hyaluronan synthase is a monomer

Cited by 4 publications

References 29 publications

Expanding glycosaminoglycan chemical space: towards the creation of sulfated analogs, novel polymers and chimeric constructs

Expanding glycosaminoglycan chemical space: towards the creation of sulfated analogs, novel polymers and chimeric constructs

Production Methods for Hyaluronan

Chemoenzymatic synthesis with the Pasteurella hyaluronan synthase; production of a multitude of defined authentic, derivatized, and analog polymers

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