Purification and Characterization of Microbial Hyaluronic Acid by Solvent Precipitation and Size-Exclusion Chromatography

Sousa, Anayla S.; Guimarães, A. P.; Gonçalves, Caroline Vieira; Silva, Ivanildo J.; Cavalcante, Célio L.; Azevedo, Diana C. S.

doi:10.1080/01496390802691281

Cited by 18 publications

(10 citation statements)

References 34 publications

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“…Hyaluronic acid is a biopolymer utilized in different current sectors of biotechnology ( Papakonstantinou et al, 2012 ; Bukhari et al, 2018 ; Bowman et al, 2018 ), and decreasing its production costs is important. Multiple reports for improvements in the production ( Liu et al, 2011 ) and recovery/purification ( Cavalcanti et al, 2019 ; Sousa et al, 2009 ; Choi et al, 2014 ; Rajendran et al, 2016 ; Akdamar et al, 2009 ; Murado et al, 2012 ; Rangaswamy and Jain, 2008 ) exist. However, a theoretical framework is required in order to determine which options provide the best production costs and have the potential to be incorporated into real-life production.…”

Section: Resultsmentioning

confidence: 99%

“…On the other hand, HA downstream bioprocessing has been extensively tested. The vast majority of the methods involved rely on a series of precipitations, filtrations, liquid-liquid partitions or chromatographic operations ( Cavalcanti et al, 2019 ; Sousa et al, 2009 ; Choi et al, 2014 ; Rajendran et al, 2016 ; Akdamar et al, 2009 ; Murado et al, 2012 ; Rangaswamy and Jain, 2008 ), each of these with specific variations to reduce material consumption or increase recovery yields.…”

Section: Introductionmentioning

confidence: 99%

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Comparative Economic Analysis Between Endogenous and Recombinant Production of Hyaluronic Acid

Torres‐Acosta

Castaneda-Aponte

Mora-Galvez

et al. 2021

Front. Bioeng. Biotechnol.

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Hyaluronic acid (HA) is a biopolymer with a wide range of applications, mainly in the cosmetic and pharmaceutical sectors. Typical industrial-scale production utilizes organisms that generate HA during their developmental cycle, such as Streptococcus equi sub. zooepidemicus. However, a significant disadvantage of using Streptococcus equi sub. zooepidemicus is that it is a zoonotic pathogen, which use at industrial scale can create several risks. This creates opportunities for heterologous, or recombinant, production of HA. At an industrial scale, the recovery and purification of HA follow a series of precipitation and filtration steps. Current recombinant approaches are developing promising alternatives, although their industrial implementation has yet to be adequately assessed. The present study aims to create a theoretical framework to forecast the advantages and disadvantages of endogenous and recombinant strains in production with the same downstream strategy. The analyses included a selection of the best cost-related recombinant and endogenous production strategies, followed by a sensitivity analysis of different production variables in order to identify the three most critical parameters. Then, all variables were analyzed by varying them simultaneously and employing multiple linear regression. Results indicate that, regardless of HA source, production titer, recovery yield and bioreactor scale are the parameters that affect production costs the most. Current results indicate that recombinant production needs to improve current titer at least 2-fold in order to compete with costs of endogenous production. This study serves as a platform to inform decision-making for future developments and improvements in the recombinant production of HA.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparative Economic Analysis Between Endogenous and Recombinant Production of Hyaluronic Acid

Torres‐Acosta

Castaneda-Aponte

Mora-Galvez

et al. 2021

Front. Bioeng. Biotechnol.

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“…Hyaluronic acid is a linear unbranched polysaccharide, composed of N-acetylglucosamine and glucuronic acid disaccharide residues [98]. It was discovered by Meyer and Palmer in 1934 in the cattle vitreous humour-the gel filling between the lens and retina of the eyeball [72].…”

Section: Hyaluronic Acidmentioning

confidence: 99%

Electrofiltration of Biopolymers

Gözke

Posten

2010

Food Eng. Rev.

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Downstream processing of bioproducts, based on the number of purification steps involved, is the most expensive part in biotechnology and food production processes. Combination of several purification steps, including membrane filtration and electrophoresis, within a single technique is known as electrofiltration-a process based on application of electric field on a dead-end filtration system parallel to the filtrate's flow direction. In this process, biopolymers, generally charged molecules with high viscosity, undergo both electrophoretic and hydrodynamic forces, reducing the thickness of the filter cake formed. As a result, electrofiltration can reduce processing time to minutes instead of hours when using conventional filtration. The objective of the present review is to assess the possibilities to purify technically and commercially important biopolymers like poly(3-hydroxybutyrate), hyaluronic acid, chitosan and xanthan by electrofiltration.

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“…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.…”

Section: Introductionmentioning

confidence: 99%

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

Nazeri

Niazi‎

Afsharifar

et al. 2021

Preprint

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Hyaluronic acid (HA), a unique polysaccharide with excellent physicochemical properties, is broadly used in pharmaceutical, biomedical, and cosmetic fields. It is widely present in all vertebrates, certain bacterial strains, and even viruses while it is not found in plants, fungi, and insects. HA is naturally synthesized by a class of integral membrane proteins called Hyaluronic acid synthase (HAS). Thus far, industrial production of HA is carried out based on either extraction from animal sources or large-scale microbial fermentation. The major drawbacks to using these systems are contamination with pathogens and microbial toxins. Recently, the production of HA through recombinant systems has received considerable attention. Plants are eco-friendly ideal expression systems for biopharmaceuticals production. In this research, the optimized human hyaluronic acid synthase2 (hHAS2) sequence was transformed into Nicotiana tabacum using Agrobacterium rhizogenes. The highest hHAS2 production in the tobacco hairy roots was 65.72 ng/kg (wet weight). The extracted HA was verified and quantified by the HA ELISA kit. The DPPH radical scavenging activity of HA with the highest concentration of 0.56 g/kg (wet weight) showed the maximum activity of 46%. Gel Permeation Chromatography (GPC) analyses revealed the high molecular weight HA with about > 0.8 MDa.

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Purification and Characterization of Microbial Hyaluronic Acid by Solvent Precipitation and Size-Exclusion Chromatography

Cited by 18 publications

References 34 publications

Comparative Economic Analysis Between Endogenous and Recombinant Production of Hyaluronic Acid

Comparative Economic Analysis Between Endogenous and Recombinant Production of Hyaluronic Acid

Electrofiltration of Biopolymers

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

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