Bacterial hyaluronic acid production through an alternative extraction method and its characterization

Abstract

BACKGROUND In this study, hyaluronic acid (HA) was produced using Streptococcus equi ssp. equi via an optimized Central Composite Design process, with purification by an alternative extraction method. Determinations of molecular weight, cytotoxicity, viscosity; proton nuclear magnetic resonance (H‐NMR) and Fourier transform infrared‐attenuated total reflection (FTIR‐ATR) analyses were provided. RESULTS The HA yield obtained was up to c. 12 g L−1 under the reaction conditions of pH 7.8, 33 °C incubation temperature, 24 h incubation time and 187 rpm agitation rate. According to gel permeation chromatography results, HA has an average molecular weight of 79 416 Da. FTIR‐ATR analysis showed that the spectrum of the standard HA sample overlapped with the spectrum obtained from this study. CONCLUSION In this study, higher titer production of HA was obtained with a simple and economical sequential process. Additionally, the results indicate that the HA produced is an appropriate biomaterial for medical applications. © 2019 Society of Chemical Industry

Applications of Hyaluronic Acid in Ophthalmology and Contact Lenses

“…Although HPLC columns are expensive, this technique is effective for analyzing HA quantitatively and qualitatively with high detection sensitivity. Other analytical techniques applied to HA analysis include Fourier transform infrared spectroscopy [ 15 , 16 , 17 ], circular dichroism [ 15 ], ultraviolet–visible absorption spectroscopy [ 15 , 16 , 17 ], and nuclear magnetic resonance spectroscopy [ 13 , 16 , 56 ]. Quantitative and qualitative methods of analyzing HA are summarized in Table 1 .…”

“…HA quantitation can be performed through a carbazole method [ 44 , 56 ], enzyme-linked immunosorbent assay [ 50 ], capillary electrophoresis [ 51 , 52 , 53 ], or HPLC, coupled with triple quadrupole mass spectrometry [ 48 ], fluorescence [ 46 ], mass spectrometric detector [ 49 , 54 ], or an ultraviolet–visible detector [ 55 ], to determine the quantification and detection limit [ 46 , 48 , 49 , 54 , 55 ]. The detection limit of capillary electrophoresis is 1-15 ppm [ 51 , 52 , 53 ].…”

Biomedical Applications of Bacterial Exopolysaccharides: A Review

“…Hyaluronic acid is produced by bacterial pathogens like Pasteurella multocida and Gram-positive Streptococcus Groups A and C [ 100 , 101 ]. Optimal bacterial growth occurred at a temperature of 37 °C and neutral pH.…”

“…However, the best HA productivity and molecular weight were observed in suboptimal growth conditions, when the growth inhibition was not connected to lower carbon uptake [ 98 ]. The carbon source can be sucrose or glucose, with the latter being the preferred source [ 101 ]. The molecular weight of HA can be enhanced by a combination of mild shear stress culture conditions and a high dissolved oxygen level [ 95 ].…”

Heterologous production of Hyaluronic Acid in Nicotiana tabacum hairy roots expressing a human Hyaluronan synthase 2

“…Thus far, HA has been extracted from animal waste, including bovine eyes and rooster combs 15-17 . Currently, this procedure is being replaced by production through bacterial fermentation [18][19][20][21][22] . These mentioned methods suffer from some serious drawbacks, including possible contamination with the animal or bacterial toxins, expensive investment in scaling-up production systems, and the timeconsuming processes of extraction and puri cation.…”