Recombinant Hyaluronic Acid Produced in Escherichia coli: Purification, Molecular-Size Profile and In Vitro Cellular Performance
7 affiliations
- 1Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Lecce, Italy
- 2Department of Bioscience, Biotechnology and Environment, University of Bari, 70125 Bari, Italy
- 3Department of Pharmacy, Health, and Nutritional Sciences, University of Calabria, 87036 Cosenza, Italy
- 4Department of Engineering for Innovation, University of Salento, Via Monteroni, 73100 Lecce, Italy
- 5Department of Molecular Microbiology, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK
- 6Department of Biophysical Analysis Platform, John Innes Centre, Norwich NR4 7UH, UK
- 7School of Pharmaceutical Sciences, Institute of Materia Medica, Xinjiang University, Urumqi 830017, China
Abstract
Recombinant production platforms are promising approaches for producing hyaluronic acid (HA) for biomedical research. In this study recombinant HA produced in Escherichia coli (rHA) was purified and compared with commercial HA (cHA) through physicochemical, rheological, and preliminary cellular analyses. Purification reduced residual protein content (from 7.47 to 0.48 mg/L), nucleic-acid-associated absorbance and endotoxins level (from 0.315 to 0.0766 EU/mg) while preserving HA recovery. Electrophoresis and hyaluronidase digestion supported HA identity and a heterogeneous apparent molecular-size profile. Mass photometry provided an exploratory, solution-based estimate of molecular-size populations, identifying a predominant native-solution population centred at approximately 194 kDa. Fourier-transform infrared spectroscopy (FT-IR) revealed characteristic HA-associated functional groups, while thermogravimetric analysis (TGA) showed an initial mass loss attributed to water evaporation (30–150 °C), followed by polymer degradation occurring at a lower temperature for rHA (~150 °C) than for cHA (~250 °C). Both samples exhibited shear-thinning behaviour, whereas rHA showed lower viscosity and moduli than cHA. In C2C12 cells, rHA produced favourable relative sulforhodamine B (SRB) signals and scratch closure responses comparable to those of the two cHA (cHA_1 and cHA_2). These findings support rHA as a purified HA-compatible material with distinctive rheological properties and preliminary in vitro cellular performance.
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BibTeX
@article{iaconisi2026recombinant,
title = {{Recombinant Hyaluronic Acid Produced in Escherichia coli: Purification, Molecular-Size Profile and In Vitro Cellular Performance}},
author = {Giorgia Natalia Iaconisi and Manuela Murano and Nunzia Gallo and Giuseppe Fiermonte and Andrew W. Truman and Natalia M. Vior and Abbas Maqbool and Roberta Del Sole and Yuan Li and Vincenza Dolce and Loredana Capobianco},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = nov,
doi = {10.3390/ijms27219997},
url = {https://www.mdpi.com/1422-0067/27/21/9997},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Recombinant Hyaluronic Acid Produced in Escherichia coli: Purification, Molecular-Size Profile and In Vitro Cellular Performance
AU - Giorgia Natalia Iaconisi
AU - Manuela Murano
AU - Nunzia Gallo
AU - Giuseppe Fiermonte
AU - Andrew W. Truman
AU - Natalia M. Vior
AU - Abbas Maqbool
AU - Roberta Del Sole
AU - Yuan Li
AU - Vincenza Dolce
AU - Loredana Capobianco
PY - 2026
DA - 2026/11/08
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27219997
UR - https://www.mdpi.com/1422-0067/27/21/9997
AB - Recombinant production platforms are promising approaches for producing hyaluronic acid (HA) for biomedical research. In this study recombinant HA produced in Escherichia coli (rHA) was purified and compared with commercial HA (cHA) through physicochemical, rheological, and preliminary cellular analyses. Purification reduced residual protein content (from 7.47 to 0.48 mg/L), nucleic-acid-associated absorbance and endotoxins level (from 0.315 to 0.0766 EU/mg) while preserving HA recovery. Electrophoresis and hyaluronidase digestion supported HA identity and a heterogeneous apparent molecular-size profile. Mass photometry provided an exploratory, solution-based estimate of molecular-size populations, identifying a predominant native-solution population centred at approximately 194 kDa. Fourier-transform infrared spectroscopy (FT-IR) revealed characteristic HA-associated functional groups, while thermogravimetric analysis (TGA) showed an initial mass loss attributed to water evaporation (30–150 °C), followed by polymer degradation occurring at a lower temperature for rHA (~150 °C) than for cHA (~250 °C). Both samples exhibited shear-thinning behaviour, whereas rHA showed lower viscosity and moduli than cHA. In C2C12 cells, rHA produced favourable relative sulforhodamine B (SRB) signals and scratch closure responses comparable to those of the two cHA (cHA_1 and cHA_2). These findings support rHA as a purified HA-compatible material with distinctive rheological properties and preliminary in vitro cellular performance.
SN - 1422-0067
ER -