scholarly journals Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production

2021 ◽  
Vol 9 (2) ◽  
pp. 312
Author(s):  
João Heitor Colombelli Manfrão-Netto ◽  
Enzo Bento Queiroz ◽  
Kelly Assis Rodrigues ◽  
Cintia M. Coelho ◽  
Hugo Costa Paes ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Acid Production ◽  
Pasteurella Multocida ◽  
Glucuronic Acid ◽  
Hansenula Polymorpha ◽  
Cultivation Conditions ◽  
Suitable Host ◽  
Genetic Switch ◽  
Glycosidic Bonds ◽  
Genes Encoding

Hyaluronic acid (HA) is a biopolymer formed by UDP-glucuronic acid and UDP-N-acetyl-glucosamine disaccharide units linked by β-1,4 and β-1,3 glycosidic bonds. It is widely employed in medical and cosmetic procedures. HA is synthesized by hyaluronan synthase (HAS), which catalyzes the precursors’ ligation in the cytosol, elongates the polymer chain, and exports it to the extracellular space. Here, we engineer Ogataea (Hansenula) polymorpha for HA production by inserting the genes encoding UDP-glucose 6-dehydrogenase, for UDP-glucuronic acid production, and HAS. Two microbial HAS, from Streptococcus zooepidemicus (hasAs) and Pasteurella multocida (hasAp), were evaluated separately. Additionally, we assessed a genetic switch using integrases in O. polymorpha to uncouple HA production from growth. Four strains were constructed containing both has genes under the control of different promoters. In the strain containing the genetic switch, HA production was verified by a capsule-like layer around the cells by scanning electron microscopy in the first 24 h of cultivation. For the other strains, the HA was quantified only after 48 h and in an optimized medium, indicating that HA production in O. polymorpha is limited by cultivation conditions. Nevertheless, these results provide a proof-of-principle that O. polymorpha is a suitable host for HA production.

scholarly journals Hyaluronic Acid Production in Bacillus subtilis

2005 ◽  
Vol 71 (7) ◽  
pp. 3747-3752 ◽  
Author(s):  
Bill Widner ◽  
Régine Behr ◽  
Steve Von Dollen ◽  
Maria Tang ◽  
Tia Heu ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Hyaluronic Acid ◽  
Acid Production ◽  
Glucuronic Acid ◽  
Hyaluronan Synthase ◽  
High Quality ◽  
Genes Encoding ◽  
High Level ◽  
Level Production ◽  
Hyaluronic Acid Production

ABSTRACT The hasA gene from Streptococcus equisimilis, which encodes the enzyme hyaluronan synthase, has been expressed in Bacillus subtilis, resulting in the production of hyaluronic acid (HA) in the 1-MDa range. Artificial operons were assembled and tested, all of which contain the hasA gene along with one or more genes encoding enzymes involved in the synthesis of the UDP-precursor sugars that are required for HA synthesis. It was determined that the production of UDP-glucuronic acid is limiting in B. subtilis and that overexpressing the hasA gene along with the endogenous tuaD gene is sufficient for high-level production of HA. In addition, the B. subtilis-derived material was shown to be secreted and of high quality, comparable to commercially available sources of HA.

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.

scholarly journals Photosynthetic conversion of CO2 to hyaluronic acid by engineered cyanobacteria

2019 ◽  
Author(s):  
Lifang Zhang ◽  
Tiago Toscano Selão ◽  
Peter J. Nixon ◽  
Birgitta Norling
Keyword(s):  
Hyaluronic Acid ◽  
Pasteurella Multocida ◽  
Glucuronic Acid ◽  
Relative Proportion ◽  
Fold Increase ◽  
Glycogen Depletion ◽  
Photosynthetic System ◽  
Photosynthetic Production ◽  
Different Types ◽  
Ha Synthase

AbstractHyaluronic acid (HA), consisting of alternating N-acetylglucosamine and glucuronic acid units, is a natural polymer with diverse cosmetic and medical applications. Currently, HA is produced by overexpressing HA synthases from gram-negative Pasteurella multocida (encoded by pmHAS) or gram-positive Streptococcus equisimilis (encoded by seHasA) in various heterotrophic microbial production platforms. Here we introduced these two different types of HA synthase into the fast-growing cyanobacterium Synechococcus sp. PCC 7002 (Syn7002) to explore the capacity for producing HA in a photosynthetic system. Our results show that both HA synthases enable Syn7002 to produce HA photoautotrophically, but that overexpression of the soluble HA synthase (PmHAS) is less deleterious to cell growth and results in higher production. Genetic disruption of the competing cellulose biosynthetic pathway increased the HA titer by over 5-fold (from 14 mg/L to 80 mg/L) and the relative proportion of HA with molecular mass greater than 2 MDa. Introduction of glmS and glmU, coding for enzymes involved in the biosynthesis of the precursor UDP-N-acetylglucosamine, in combination with partial glycogen depletion, allowed photosynthetic production of 112 mg/L of HA in 5 days, an 8-fold increase in comparison to the initial PmHAS expressing strain. Addition of tuaD and gtaB (coding for genes involved in UDP-glucuronic acid biosynthesis) also improved the HA yield, albeit to a lesser extent. Overall our results have shown that cyanobacteria hold promise for sustainable production of pharmaceutically important polysaccharides from sunlight and CO2.

scholarly journals Advantages of Hyaluronic Acid and Its Combination with Other Bioactive Ingredients in Cosmeceuticals

Molecules ◽  
2021 ◽  
Vol 26 (15) ◽  
pp. 4429
Author(s):  
Anca Maria Juncan ◽  
Dana Georgiana Moisă ◽  
Antonello Santini ◽  
Claudiu Morgovan ◽  
Luca-Liviu Rus ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Glucuronic Acid ◽  
Biological Effects ◽  
Biological Processes ◽  
Cosmetic Products ◽  
Aesthetic Medicine ◽  
Cosmetic Formulations ◽  
Other Substances ◽  
Bioactive Ingredients ◽  
Brand Portfolio

This study proposes a review on hyaluronic acid (HA) known as hyaluronan or hyaluronate and its derivates and their application in cosmetic formulations. HA is a glycosaminoglycan constituted from two disaccharides (N-acetylglucosamine and D-glucuronic acid), isolated initially from the vitreous humour of the eye, and subsequently discovered in different tissues or fluids (especially in the articular cartilage and the synovial fluid). It is ubiquitous in vertebrates, including humans, and it is involved in diverse biological processes, such as cell differentiation, embryological development, inflammation, wound healing, etc. HA has many qualities that recommend it over other substances used in skin regeneration, with moisturizing and anti-ageing effects. HA molecular weight influences its penetration into the skin and its biological activity. Considering that, nowadays, hyaluronic acid has a wide use and a multitude of applications (in ophthalmology, arthrology, pneumology, rhinology, aesthetic medicine, oncology, nutrition, and cosmetics), the present study describes the main aspects related to its use in cosmetology. The biological effect of HA on the skin level and its potential adverse effects are discussed. Some available cosmetic products containing HA have been identified from the brand portfolio of most known manufacturers and their composition was evaluated. Further, additional biological effects due to the other active ingredients (plant extracts, vitamins, amino acids, peptides, proteins, saccharides, probiotics, etc.) are presented, as well as a description of their possible toxic effects.

Effects of glucuronic acid and N-acetyl-D-glucosamine supplementation on the perivitelline space during the IVM of pig oocytes

2020 ◽  
Vol 32 (10) ◽  
pp. 941
Author(s):  
J. Z. Current ◽  
B. D. Whitaker
Keyword(s):  
Hyaluronic Acid ◽  
Glutathione Peroxidase ◽  
Embryo Development ◽  
Glucuronic Acid ◽  
The Other ◽  
Perivitelline Space

The objective of this study was to minimise polyspermic penetration by increasing the perivitelline space (PVS) thickness through supplementation of the hyaluronic acid components glucuronic acid and N-acetyl-d-glucosamine (GlcNAc). Oocytes (n=4690) were supplemented during the first 24h and/or the remainder of maturation (final 16–18h) with 0.01mM glucuronic acid and 0.01mM GlcNAc and then evaluated for PVS thickness, hyaluronic acid, glutathione and glutathione peroxidase concentrations. Fertilised oocytes were evaluated for polyspermic penetration and embryo development. The PVS thickness and amount of hyaluronic acid was significantly (P<0.05) greater in oocytes supplemented with 0.01mM glucuronic acid and 0.01mM GlcNAc during the second part or all of maturation compared with the other treatments. In addition, polyspermic penetration was significantly (P<0.05) less in oocytes supplemented with 0.01mM glucuronic acid and 0.01mM GlcNAc during the second part or all of maturation compared with the other treatments. Supplementing 0.01mM glucuronic acid and GlcNAc during maturation significantly (P<0.05) increased the percentage of cleaved embryos by 48h after IVF and blastocysts formed by 144h after IVF compared those not supplemented. These results indicate that supplementing PVS components during maturation decreases polyspermic penetration by increasing PVS thickness.

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