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 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 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 Influence of Phytohormones on Monosaccharide Composition of Polysaccharides from Wheat Suspension Culture

2017 ◽  
Vol 19 (3) ◽  
pp. 231 ◽  
Author(s):  
N. Bishimbayeva ◽  
A. Murtazina ◽  
G. McDougall
Keyword(s):  
Chemical Composition ◽  
Suspension Culture ◽  
Culture Medium ◽  
Glucuronic Acid ◽  
Physiological Activity ◽  
Fold Increase ◽  
Monosaccharide Composition ◽  
Medium Composition ◽  
Plant Polysaccharides ◽  
Different Types

Plant polysaccharides with technical and physiologic traits attract researchers by their high physiological activity in regulation of the growth, development and protective reactions. Cell cultures allow to regulate chemical composition of synthesized substances by changing media composition and are widely used to enhance or change the biosynthesis of metabolites. The aim of this study was to investigate the influence of phytohormones 2,4-dichlorphenoxyacetic acid (2,4 –D) and abscisic acid (ABA) of culture medium on chemical composition of polysaccharides (PS), extracted from cells and extracellular liquid of wheat suspension culture. It was shown for the medium with ABA that monosaccharide composition of extracellular PS mainly represented by glucose (87%), whereas PS isolated from cells were rich for xylose and glucuronic acid. Monosaccharide composition of extracellular PS from media with 2,4-D showed 6-fold increase of arabinose, 8-fold ‒ of galactose, 5-fold ‒ of xylose and glucuronic acid, compared to extracellular PS from ABA medium. Composition of cellular PS from media with 2,4-D were mainly similar to ABA and differed by the increased amount of mannose (3-fold), and galacturonic acid (2,5-fold). Thus, regulative effect of the use of two different types of phytohormones was demonstrated on the biosynthesis of variously composed polysaccharides.

scholarly journals HYBID (alias KIAA1199/CEMIP) and hyaluronan synthase coordinately regulate hyaluronan metabolism in histamine-stimulated skin fibroblasts

2020 ◽  
Vol 295 (8) ◽  
pp. 2483-2494
Author(s):  
Hiroyuki Yoshida ◽  
Mika Aoki ◽  
Aya Komiya ◽  
Yoko Endo ◽  
Keigo Kawabata ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Fold Increase ◽  
Histamine Receptor ◽  
Skin Fibroblasts ◽  
Dependent Manner ◽  
Skin Damage ◽  
Photoaged Skin ◽  
Molecular Sizes ◽  
Ha Synthase ◽  
Hyaluronan Binding

The immune-regulatory compound histamine is involved in the metabolism of the essential skin component hyaluronan (HA). We previously reported that histamine up-regulates the expression of HYBID (hyaluronan-binding protein involved in hyaluronan depolymerization, also called CEMIP or KIAA1199), which plays a key role in HA degradation. However, no information is available about histamine's effects on HA synthase (HAS) expression, the molecular sizes of HA species produced, and histamine receptors and their signaling pathways in skin fibroblasts. Moreover, histamine's effects on photoaged skin remain elusive. Here, we show that histamine increases HA degradation by up-regulating HYBID and down-regulating HAS2 in human skin fibroblasts in a dose- and time-dependent manner and thereby decreases the total amounts and sizes of newly produced HA. Histamine H1 blocker abrogated the histamine effects on HYBID up-regulation, HAS2 suppression, and HA degradation. Histamine H1 agonist exhibited effects on HA levels, composition, and breakdown similar to those of histamine. Of note, blockade of protein kinase Cδ or PI3K–Akt signaling abolished histamine-mediated HYBID stimulation and HAS2 suppression, respectively. Immunohistochemical experiments revealed a significant ∼2-fold increase in tryptase-positive mast cells in photoaged skin, where HYBID and HAS2 expression levels were increased and decreased, respectively, compared with photoprotected skin. These results indicate that histamine controls HA metabolism by up-regulating HYBID and down-regulating HAS2 via distinct signaling pathways downstream of histamine receptor H1. They further suggest that histamine may contribute to photoaged skin damage by skewing HA metabolism toward degradation.

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.

scholarly journals Inhibitor of Hyaluronic Acid Synthesis 4-Methylumbelliferone as an Anti-Inflammatory Modulator of LPS-Mediated Astrocyte Responses

2020 ◽  
Vol 21 (21) ◽  
pp. 8203 ◽  
Author(s):  
Dmitry V. Chistyakov ◽  
Arina I. Nikolskaya ◽  
Sergei V. Goriainov ◽  
Alina A. Astakhova ◽  
Marina G. Sergeeva
Keyword(s):  
Hyaluronic Acid ◽  
Inflammatory Responses ◽  
Specific Inhibitor ◽  
Acid Synthesis ◽  
Interleukin 10 ◽  
Ultra Performance Liquid Chromatography ◽  
Necrosis Factor Alpha ◽  
Inflammatory Stimuli ◽  
Anti Inflammatory ◽  
Ha Synthase

Astrocytes are glial cells that play an important role in neuroinflammation. Astrocytes respond to many pro-inflammatory stimuli, including lipopolysaccharide (LPS), an agonist of Toll-like receptor 4 (TLR4). Regulatory specificities of inflammatory signaling pathways are still largely unknown due to the ectodermal origin of astrocytes. Recently, we have shown that hyaluronic acid (HA) may form part of astrocyte inflammatory responses. Therefore, we tested 4-methylumbelliferone (4-MU), a specific inhibitor of HA synthesis, as a possible regulator of LPS-mediated responses. Rat primary astrocytes were treated with LPS with and without 4-MU and gene expression levels of inflammatory (interleukins 1β, (IL-1β), 6, (IL-6), tumor necrosis factor alpha TNFα,) and resolution interleukin 10 (IL-10) markers were evaluated via real-time PCR and western blot. The release of cytokines and HA was determined by ELISA. Oxylipin profiles were measured by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. Our data show that 4-MU (i) has anti-inflammatory effects in the course of TLR4 activation, decreasing the cytokines level TNFα, IL-6 and IL-1β and increasing IL-10, (ii) downregulates prostaglandin synthesis but not via cyclooxygenases COX-1 and COX-2 pathways, (iii) modulates HA synthesis and decreases LPS-induced HA synthase mRNA expression (HAS-1, HAS-2) but does not have an influence on HAS-3, HYAL1 and HYAL2 mRNAs; (iv) the effects of 4-MU are predominantly revealed via JNK but not p38, ERK mitogen-activated protein kinases (MAPKs) or nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) pathways. For the first time, it is shown that 4-MU possesses the useful potential to regulate an inflammatory astrocyte response.

Far-ultraviolet circular dichroism of N-acetylglucosamine, glucuronic acid, and hyaluronic acid

1977 ◽  
Vol 99 (6) ◽  
pp. 1730-1734 ◽  
Author(s):  
Lynn A. Buffington ◽  
Eugene S. Pysh ◽  
Bireswar Chakrabarti ◽  
Endre A. Balazs
Keyword(s):  
Hyaluronic Acid ◽  
Circular Dichroism ◽  
Glucuronic Acid ◽  
Far Ultraviolet ◽  
Circular Dichroïsm

scholarly journals Preparation, Characterization, and Inhibition of Hyaluronic Acid Oligosaccharides in Triple-Negative Breast Cancer

Biomolecules ◽  
2019 ◽  
Vol 9 (9) ◽  
pp. 436 ◽  
Author(s):  
Wenwei Han ◽  
Lili Song ◽  
Yingdi Wang ◽  
Youjing Lv ◽  
Xiangyan Chen ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Glucuronic Acid ◽  
Biological Function ◽  
Critical Component ◽  
Potential Value ◽  
And Migration

Hyaluronic acid (hyaluronan, HA) is a critical component of the extracellular matrix and plays an important biological function of interacting with different molecules and receptors. In this study, both odd- and even-numbered HA oligosaccharides (HAOs) with specific degrees of polymerization (DP) were prepared by different hydrochloric acid hydrolyses, and their structures were characterized by means of HPLC, ESI-MS, and NMR. The data show that the odd-numbered HAOs (DP3-11) have a glucuronic acid reducing end, while the even-numbered HAOs (DP2-10) have an N-acetylglucosamine reducing end. Biological evaluations indicated that all HAOs significantly inhibited the growth and migration of triple-negative breast cancer (TNBC) MDA-MB-231 cells. Among these oligosaccharides, the HA tetrasaccharide (DP4) was confirmed to be the minimum fragment necessary to inhibit MDA-MB-231 cells. Our data suggest that HAOs have potential value in the treatment of TNBC.

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