Production of isotopically enriched high molecular weight hyaluronic acid and characterization by solid-state NMR

Rampratap, Pushpa; Lasorsa, Alessia; Perrone, Barbara; Wel, Patrick C.A. van der; Walvoort, Marthe T. C.

doi:10.1016/j.carbpol.2023.121063

Cited by 8 publications

(9 citation statements)

References 49 publications

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“…S. equi subsp. zooepidemicus bacteria 29 . The HA molecules used in this study have a 13 C enrichment of ∼96 % and an average MW ∼275 kDa 29 .…”

Section: Resultsmentioning

confidence: 99%

“…The 13 C 1D INEPT spectrum showed good signal to noise ratio (Figure 2A). The resonances were initially assigned based on published solution and ssNMR data 29,30 , however more peaks were detected compared to the number of HA carbons (Figure 1B). To investigate these unexpected signals in more detail, we performed multidimensional ssNMR experiments.…”

Section: Resultsmentioning

confidence: 99%

“…The 13 C enriched HMW-HA was produced via an optimized protocol, as described in previous work 28 . Briefly, Streptococcus equi subspecies zooepidemicus (DSM 20727, obtained from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures), were subcultured and grown by using two percent inoculums containing 3% (w/v) TSB (tryptone soy broth) and 1% glucose.…”

Section: Production and Purification Of 13 C-labelled Hmw-hamentioning

confidence: 99%

“…To enable a detailed analysis of HMW-HA by ssNMR, we produced 13 C-labeled HMW-HA using S. equi subsp. zooepidemicus bacteria 28 . The HA molecules used in this study have a 13 C enrichment of ~96 % and an average MW ~275 kDa 28 .…”

Section: Atom-specific Analysis Of Hydrated 13 C-labeled Hmw-ha By Ma...mentioning

confidence: 99%

“…zooepidemicus bacteria 28 . The HA molecules used in this study have a 13 C enrichment of ~96 % and an average MW ~275 kDa 28 . Our initial focus was on elucidating the effect of hydration on the biopolymers, given the fact that the hydration behavior of HA is one of its key functional properties.…”

Section: Atom-specific Analysis Of Hydrated 13 C-labeled Hmw-ha By Ma...mentioning

confidence: 99%

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Resolving atomic-level dynamics and interactions of high molecular weight hyaluronic acid by multidimensional solid-state NMR

Rampratap,

Lasorsa,

Arunachalam

et al. 2024

Preprint

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High molecular weight (HMW) hyaluronic acid (HA) is a highly abundant natural polysaccharide and a fundamental component of the extracellular matrix (ECM). Its size and concentration regulate tissues’ macro– and microenvironments, and its upregulation is a hallmark feature of certain tumors. Yet, the conformational dynamics of HMW-HA and how it engages with components of the ECM microenvironment remain poorly understood on the molecular level. Probing the molecular structure and dynamics of HMW polysaccharides in a hydrated, physiological-like environment is crucial but also technically challenging. Here, we deploy advanced magic-angle-spinning (MAS) solid-state NMR (ssNMR) spectroscopy in combination with isotopic enrichment to enable an in-depth study of HMW-HA to address this challenge. This approach resolves multiple coexisting HA conformations and dynamics as a function of environmental conditions. By combining13C-labeled HA with unlabeled ECM components, we detect by MAS NMR HA-specific changes in global and local conformational dynamics as a consequence of environmental conditions. These measurements reveal atom-specific variations in dynamics and structure of the N-acetylglucosamine (GlcNAc) moiety of HA. We discuss possible implications for interactions that stabilize the structure of HMW-HA and facilitate its recognition by HA-binding proteins. The described methods apply similarly to studies of the molecular structure and dynamics of HA in tumor contexts and in other biological tissues, as well as HMW-HA hydrogels and nanoparticles used for biomedical and/or pharmaceutical applications.

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“…S. equi subsp. zooepidemicus bacteria 29 . The HA molecules used in this study have a 13 C enrichment of ∼96 % and an average MW ∼275 kDa 29 .…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Production and Purification Of 13 C-labelled Hmw-hamentioning

confidence: 99%

Section: Atom-specific Analysis Of Hydrated 13 C-labeled Hmw-ha By Ma...mentioning

confidence: 99%

Section: Atom-specific Analysis Of Hydrated 13 C-labeled Hmw-ha By Ma...mentioning

confidence: 99%

See 3 more Smart Citations

Resolving atomic-level dynamics and interactions of high molecular weight hyaluronic acid by multidimensional solid-state NMR

Rampratap,

Lasorsa,

Arunachalam

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Hyaluronic acid-based hydrogels as codelivery systems: The effect of intermolecular interactions investigated by HR-MAS and solid-state NMR Spectroscopy

Vanoli,

Casalegno,

Carravetta

et al. 2025

Carbohydrate Polymers

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Hyaluronic acid application strategies for plant bioactive component delivery: A review

Zhang,

Huang,

Liu

et al. 2024

International Journal of Biological Macromolecules

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Production of isotopically enriched high molecular weight hyaluronic acid and characterization by solid-state NMR

Cited by 8 publications

References 49 publications

Resolving atomic-level dynamics and interactions of high molecular weight hyaluronic acid by multidimensional solid-state NMR

Resolving atomic-level dynamics and interactions of high molecular weight hyaluronic acid by multidimensional solid-state NMR

Hyaluronic acid-based hydrogels as codelivery systems: The effect of intermolecular interactions investigated by HR-MAS and solid-state NMR Spectroscopy

Hyaluronic acid application strategies for plant bioactive component delivery: A review

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