Characterization of UDP-glucose dehydrogenase from Pasteurella multocida CVCC 408 and its application in hyaluronic acid biosynthesis

2016 ◽  
Vol 85 ◽  
pp. 64-70 ◽  
Author(s):  
Xin Chu ◽  
Jian Han ◽  
Donghui Guo ◽  
Zhenzhou Fu ◽  
Weifeng Liu ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Pasteurella Multocida ◽  
Glucose Dehydrogenase ◽  
Acid Biosynthesis

scholarly journals Heterologous Hyaluronic Acid Production in Kluyveromyces lactis

2019 ◽  
Vol 7 (9) ◽  
pp. 294
Author(s):  
Antonio M. V. Gomes ◽  
João H. C. M. Netto ◽  
Lucas S. Carvalho ◽  
Nádia S. Parachin
Keyword(s):  
Hyaluronic Acid ◽  
Pathogenic Bacteria ◽  
Pasteurella Multocida ◽  
Glucuronic Acid ◽  
Kluyveromyces Lactis ◽  
Glucose Dehydrogenase ◽  
Primary Sources ◽  
Batch Mode ◽  
Dehydrogenase Gene ◽  
Ha Synthase

Hyaluronic Acid (HA) is a biopolymer composed by the monomers Glucuronic Acid (GlcUA) and N-Acetyl Glucosamine (GlcNAc). It has a broad range of applications in the field of medicine, being marketed between USD 1000–5000/kg. Its primary sources include extraction of animal tissue and fermentation using pathogenic bacteria. However, in both cases, extensive purification protocols are required to prevent toxin contamination. In this study, aiming at creating a safe HA producing microorganism, the generally regarded as safe (GRAS) yeast Kluyveroymyces lactis is utilized. Initially, the hasB (UDP-Glucose dehydrogenase) gene from Xenopus laevis (xlhasB) is inserted. After that, four strains are constructed harboring different hasA (HA Synthase) genes, three of humans (hshasA1, hshasA2, and hshasA3) and one with the bacteria Pasteurella multocida (pmhasA). Transcript values analysis confirms the presence of hasA genes only in three strains. HA production is verified by scanning electron microscopy in the strain containing the pmHAS isoform. The pmHAS strain is grown in a 1.3 l bioreactor operating in a batch mode, the maximum HA levels are 1.89 g/L with a molecular weight of 2.097 MDa. This is the first study that reports HA production in K. lactis and it has the highest HA titers reported among yeast.

The isolation and characterization of hyaluronic acid from pasteurella multocida

1970 ◽  
Vol 14 (2) ◽  
pp. 272-276 ◽  
Author(s):  
J.A. Cifonelli ◽  
P.A. Rebers ◽  
K.H. Heddleston
Keyword(s):  
Hyaluronic Acid ◽  
Pasteurella Multocida ◽  
Isolation And Characterization

Preparation and Characterization of the High Molecular Weight [3H]Hyaluronic Acid

1992 ◽  
Vol 57 (10) ◽  
pp. 2151-2156 ◽  
Author(s):  
Peter Chabreček ◽  
Ladislav Šoltés ◽  
Hynek Hradec ◽  
Jiří Filip ◽  
Eduard Orviský
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
High Molecular Weight ◽  
Methyl Bromide ◽  
High Performance ◽  
Hydrogen Atoms ◽  
Isolation And Characterization ◽  
Labelling Procedure ◽  
Enzymatic Depolymerization

Two methods for the preparation of high molecular weight [3H]hyaluronic acid were investigated. In the first one, hydrogen atoms in the molecule were replaced by tritium. This isotopic substitution was performed in aqueous solution using Pd/CaCO3 as the catalyst. In the second method, the high molecular weight hyaluronic acid was alkylated with [3H]methyl bromide in liquid ammonia at a temperature of -33.5 °C. High-performance gel permeation chromatographic separation method was used for the isolation and characterization of the high molecular weight [3H]hyaluronic acid. Molecular weight parameters for the labelled biopolymers were Mw = 128 kDa, Mw/Mn = 1.88 (first method) and Mw = 268 kDa, Mw/Mn = 1.55 (second method). The high molecular weight [3H]hyaluronic acid having Mw = 268 kDa was degraded further by specific hyaluronidase. Products of the enzymatic depolymerization were observed to be identical for both, labelled and cold biopolymer. This finding indicates that the described labelling procedure using [3H]methyl bromide does not induce any major structural rearrangements in the molecule.

scholarly journals Toward Physicochemical and Rheological Characterization of Different Injectable Hyaluronic Acid Dermal Fillers Cross-Linked with Polyethylene Glycol Diglycidyl Ether

Polymers ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 948
Author(s):  
Nicola Zerbinati ◽  
Sabrina Sommatis ◽  
Cristina Maccario ◽  
Maria Chiara Capillo ◽  
Giulia Grimaldi ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Polyethylene Glycol ◽  
Biological Activities ◽  
Diglycidyl Ether ◽  
Dermal Filler ◽  
Cross Linking ◽  
Matrix Structure ◽  
Dermal Fillers ◽  
Rheological Characterization

(1) Background: Injectable hyaluronic acid (HA) dermal fillers are used to restore volume, hydration and skin tone in aesthetic medicine. HA fillers differ from each other due to their cross-linking technologies, with the aim to increase mechanical and biological activities. One of the most recent and promising cross-linkers is polyethylene glycol diglycidyl ether (PEGDE), used by the company Matex Lab S.p.A., (Brindisi, Italy) to create the HA dermal filler PEGDE family. Over the last few years, several studies have been performed to investigate the biocompatibility and biodegradability of these formulations, but little information is available regarding their matrix structure, rheological and physicochemical properties related to their cross-linking technologies, the HA content or the degree of cross-linking. (2) Methods: Seven different injectable HA hydrogels were subjected to optical microscopic examination, cohesivity evaluation and rheological characterization in order to investigate their behavior. (3) Results: The analyzed cross-linked dermal fillers showed a fibrous “spiderweb-like” matrix structure, with each medical device presenting different and peculiar rheological features. Except for HA non cross-linked hydrogel 18 mg/mL, all showed an elastic and cohesive profile. (4) Conclusions: The comparative analysis with other literature works makes a preliminary characterization of these injectable medical devices possible.

scholarly journals Characterization of proteoglycan and the proteoglycan-hyaluronic acid complex by electric birefringence

1978 ◽  
Vol 173 (1) ◽  
pp. 237-243 ◽  
Author(s):  
M Isles ◽  
A R Foweraker ◽  
B R Jennings ◽  
T Hardingham ◽  
H Muir
Keyword(s):  
Hyaluronic Acid ◽  
Particle Orientation ◽  
Dilute Solution ◽  
Relaxation Rates ◽  
Acid Complex ◽  
Electric Birefringence ◽  
Partial Alignment ◽  
Uronic Acid Content ◽  
Birefringence Relaxation

An electric field causes partial alignment of macromolecules in a dilute solution. The accompanying changes in the solution birefringence offer a sensitive and quick means of monitoring the rates of particle orientation and hence the size of the solute molecules. Such measurements are reported for dilute solutions of proteoglycans in the absence and presence of added hyaluronic acid. The proteoglycan molecules are shown to be some 580 nm long. In the presence of hyaluronic acid they form aggregates that appear to be consistent with the model previously proposed in which the proteoglycans attach radially to the extended hyaluronic acid chain. The electric-birefringence relaxation rates indicate aggregates of similar length to that of the extended hyaluronic acid chain, with the proteoglycans spaced on average at 29nm intervals. A proteoglycan sample the cystine residues of which had been reduced and alkylated showed no evidence of aggregation with hyaluronic acid up to the concentrations of the acid corresponding to 1% of the total uronic acid content. The electric-birefringence method is shown to have a large potential in the study of associating polysaccharide solutions.

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