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Downstream process intensification for biotechnologically generated hyaluronic acid: Purification and
Priya Shukla1, Pradeep Srivastava1, Abha Mishra1
1School of Biochemical Engineering, Indian Institute of Technology (Banaras Hindu University), 221005 Varanasi, India.
Journal of Bioscience and Bioengineering
|July 1, 2023
Summary
This study presents an effective method for purifying hyaluronic acid (HA) from fermented broth. The optimized process yields high-purity HA suitable for pharmaceutical applications.
Area of Science:
- Biochemistry
- Biotechnology
- Materials Science
Background:
- Hyaluronic acid (HA), a glycosaminoglycan, possesses diverse clinical applications.
- Efficient purification methods are crucial for maximizing HA recovery and purity.
- Streptococcus zooepidemicus fermentation is a common source for microbial HA production.
Purpose of the Study:
- To investigate and optimize downstream purification techniques for hyaluronic acid.
- To achieve maximum recovery and high purity of HA from fermented broth.
- To validate the purity and structural integrity of the purified HA.
Main Methods:
- Fermentation of Streptococcus zooepidemicus MTCC 3523 to produce HA.
- Multi-step purification involving filtration, adsorption (activated carbons, XAD-7 resins), and diafiltration.
- Characterization using Fourier transform-infrared spectroscopy, X-ray diffraction, nuclear magnetic resonance, and scanning electron microscopy.
- Antioxidant activity assays including DPPH radical-scavenging, total antioxidant capacity, hydroxyl radical-scavenging, and reducing power.
Main Results:
- Nucleic acids and high molecular weight proteins were removed using activated carbons and XAD-7 resins.
- Insoluble and low molecular weight impurities were effectively cleared by diafiltration.
- Achieved HA recovery of 79.16% with purity approaching 90%.
- Validated HA presence, purity, and structure through various analytical techniques.
- Demonstrated significant antioxidant activities: DPPH radical-scavenging (4.87 ± 0.45 kmol TE/g), total antioxidant capacity (13.32 ± 0.52%), hydroxyl radical-scavenging (32.03 ± 0.12%), and reducing power (24.85 ± 0.45%).
Conclusions:
- Precipitation, adsorption, and diafiltration are effective methods for HA extraction from fermented broth.
- The optimized process yields pharmaceutical-grade hyaluronic acid suitable for non-injectable applications.
- The purified HA exhibits notable antioxidant properties.
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