Performance of the main downstream operations on hyaluronic acid purification

Cavalcanti, A. D. D.; Ferreira, Bruno Armenio Moreira; Santana, Maria Helena Andrade

doi:10.1016/j.procbio.2020.08.020

Cited by 16 publications

(7 citation statements)

References 117 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For microbial HA, the downstream process often involves a combination of alcohol precipitation, adsorption on silica gel, and/or activated charcoal and diafiltration steps (Patil et al 2011 ; Rangaswamy and Jain 2008 ). Proteins are the main impurities present in microbial HA (Cavalcanti et al 2020 ), but endotoxins from pathogenic bacteria, such as S. zooepidemicus , also represent potential safety concerns (Liu et al 2011 ). Obtaining highly purified HA is a challenging task, and given its increasing market demand, there is an urgent need for the development of a more efficient purification process.…”

Section: Introductionmentioning

confidence: 99%

Primary recovery of hyaluronic acid produced in Streptococcus equi subsp. zooepidemicus using PEG–citrate aqueous two-phase systems

et al. 2021

View full text Add to dashboard Cite

Given its biocompatibility, rheological, and physiological properties, hyaluronic acid (HA) has become a biomaterial of increasing interest with multiple applications in medicine and cosmetics. In recent decades, microbial fermentations have become an important source for the industrial production of HA. However, due to its final applications, microbial HA must undergo critical and long purification processes to ensure clinical and cosmetic grade purity. Aqueous two-phase systems (ATPS) have proven to be an efficient technique for the primary recovery of high-value biomolecules. Nevertheless, their implementation in HA downstream processing has been practically unexplored. In this work, polyethylene glycol (PEG)–citrate ATPS were used for the first time for the primary recovery of HA produced with an engineered strain of Streptococcus equi subsp. zooepidemicus. The effects of PEG molecular weight (MW), tie-line length (TLL), volume ratio (VR), and sample load on HA recovery and purity were studied with a clarified fermentation broth as feed material. HA was recovered in the salt-rich bottom phase, and its recovery increased when a PEG MW of 8000 g mol−1 was used. Lower VR values (0.38) favoured HA recovery, whereas purity was enhanced by a high VR (3.50). Meanwhile, sample load had a negative impact on both recovery and purity. The ATPS with the best performance was PEG 8000 g mol−1, TLL 43% (w/w), and VR 3.50, showing 79.4% HA recovery and 74.5% purity. This study demonstrated for the first time the potential of PEG–citrate ATPS as an effective primary recovery strategy for the downstream process of microbial HA.

show abstract

Section: Introductionmentioning

confidence: 99%

Primary recovery of hyaluronic acid produced in Streptococcus equi subsp. zooepidemicus using PEG–citrate aqueous two-phase systems

et al. 2021

View full text Add to dashboard Cite

show abstract

“…The recovered precipitates were redissolved in distilled water and dialyzed against distilled water to remove inorganic salts. The dialyzed solutions were treated with activated charcoal and then passed through ion exchange resin based on the method for purification of fermented hyaluronan 22 . Chondroitins in the eluent were obtained as white powders by ethanol precipitation and subsequent vacuum drying.…”

Section: Methodsmentioning

confidence: 99%

“…The dialyzed solutions were treated with activated charcoal and then passed through ion exchange resin based on the method for purification of fermented hyaluronan. 22 Chondroitins in the eluent were obtained as white powders by ethanol precipitation and subsequent vacuum drying. The weight-average molecular weight and the number-average molecular weight of chondroitins were determined by size exclusion chromatography (Ultrahydrogel Linear, 7.8 Â 150 mm, Waters Corporation).…”

Section: Preparation Of Unlabeled and Isotopically Labeled Chondroitinsmentioning

confidence: 99%

NMR characterization of uniformly ¹³C‐ and/or ¹⁵N‐labeled, unsulfated chondroitins with high molecular weights

Ichikawa,

Otsuka,

Minamisawa

et al. 2024

Magnetic Reson in Chemistry

View full text Add to dashboard Cite

Solution nuclear magnetic resonance (NMR) analysis of polysaccharides can provide valuable information not only on their primary structures but also on their conformation, dynamics, and interactions under physiological conditions. One of the main problems is that non‐anomeric 1H signals typically overlap, and this often hinders detailed NMR analysis. Isotope enrichment, such as with 13C and 15N, will add a new dimension to the NMR spectra of polysaccharides, and spectral analysis can be performed with enhanced sensitivity using isolated peaks. For this purpose, here we have prepared uniformly 13C‐ and/or 15N‐labeled chondroitin polysaccharides –4)‐β‐D‐glucuronopyranosyl‐(1–3)‐2‐acetamido‐2‐deoxy‐β‐D‐galactopyranosyl‐(1– with molecular weights in the range from 310 to 460 k by bacterial fermentation. The enrichment ratios for 13C and 15N were 98.9 and 99.8%, respectively, based on the mass spectrometric analysis of the constituent chondroitin disaccharides. 1H and 13C NMR signals were assigned mainly based on HSQC and 13C‐detection experiments including INADEQUATE, HETCOR, and HETCOR‐TOCSY. The carbonyl carbon signal of the N‐acetyl‐β‐D‐galactosamine residue was unambiguously distinguished from the C6 carbon of the β‐D‐glucuronic acid residue by the observation of 13C peak splitting due to 1JCN coupling in 13C‐ and 15N‐labeled chondroitin. The T2* and T1 were measured and indicate that both rigid and mobile sites are present in the long sequence of chondroitin. The conformation, dynamics, and interactions of chondroitin and its derivatives will be further analyzed based on the results obtained in this study.

show abstract

“…Key assumptions regarding the Upstream Section, such as medium composition, fermentation duration, and yield of HA on substrate were based on laboratory scale data previously published [29]. The primary recovery and purification sections were based on the scientific and patent literature, especially references [5,21,30,31]. A total of four process scenarios were created: S1, S2, S3, and S4.…”

Section: Design Basis and Process Scenariosmentioning

confidence: 99%

“…HA's properties are closely related to its molecular weight (MW), which typically ranges from 10 3 to 10 7 Da [5]. Due to its large MW and capacity to establish multiple hydrogen bonds with water molecules, HA is very hygroscopic and exhibits unique rheological and adhesive properties.…”

Section: Introductionmentioning

confidence: 99%

Techno-Economic Analysis of a Hyaluronic Acid Production Process Utilizing Streptococcal Fermentation

et al. 2021

Self Cite

View full text Add to dashboard Cite

Hyaluronic acid (HA) is a polysaccharide of alternating d-glucuronic acid and N-acetyl-d-glucosamine residues present in the extracellular matrix of connective, epithelial, and nervous tissues. Due to its singular hydrating, rheological and adhesive properties, HA has found numerous cosmetic and medical applications. However, techno-economic analyses of high value-added bioproducts such as HA are scarce in the literature. Here, we present a techno-economic analysis of a process for producing HA using Streptococcus zooepidemicus, simulated in SuperPro Designer. In the baseline scenario, HA is produced by batch fermentation, reaching 2.5 g/L after 24 h. It is then centrifuged, diafiltered, treated with activated carbon and precipitated with isopropanol. The product is suitable for topical formulations and its production cost was estimated as 1115 $/kg. A similar scenario, based on fed-batch culture and assuming a titer of 5.0 g/L, led to a lower cost of 946 $/kg. Moreover, in two additional scenarios, 10% of the precipitated HA is diverted to the production of a highly pure and high-molecular weight HA, suitable for injectable applications. These scenarios resulted in higher capital and operating costs, but also in higher profits, because HA for injectable use has a higher selling price that more than compensates for its higher production costs.

show abstract

Performance of the main downstream operations on hyaluronic acid purification

Cited by 16 publications

References 117 publications

Primary recovery of hyaluronic acid produced in Streptococcus equi subsp. zooepidemicus using PEG–citrate aqueous two-phase systems

Primary recovery of hyaluronic acid produced in Streptococcus equi subsp. zooepidemicus using PEG–citrate aqueous two-phase systems

NMR characterization of uniformly ¹³C‐ and/or ¹⁵N‐labeled, unsulfated chondroitins with high molecular weights

Techno-Economic Analysis of a Hyaluronic Acid Production Process Utilizing Streptococcal Fermentation

Contact Info

Product

Resources

About

Performance of the main downstream operations on hyaluronic acid purification

Cited by 16 publications

References 117 publications

Primary recovery of hyaluronic acid produced in Streptococcus equi subsp. zooepidemicus using PEG–citrate aqueous two-phase systems

Primary recovery of hyaluronic acid produced in Streptococcus equi subsp. zooepidemicus using PEG–citrate aqueous two-phase systems

NMR characterization of uniformly 13C‐ and/or 15N‐labeled, unsulfated chondroitins with high molecular weights

Techno-Economic Analysis of a Hyaluronic Acid Production Process Utilizing Streptococcal Fermentation

Contact Info

Product

Resources

About

NMR characterization of uniformly ¹³C‐ and/or ¹⁵N‐labeled, unsulfated chondroitins with high molecular weights