Current state on the enzymatic synthesis of glycosaminoglycans

Gottschalk, Johannes; Elling, Lothar

doi:10.1016/j.cbpa.2020.09.008

Cited by 33 publications

(28 citation statements)

References 69 publications

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“…[6][7][8][9] Therefore, enzymatic strategies to produce HA are the focus of research. [10,11] In our previous study, we established an in vitro one-pot synthesis to produce high molecular weight (HMW) HA. [12] The enzymatic cascade contains six enzymes: AtGlcAK, AtUSP (both from Arabidopsis thaliana), BlNahK (Bifidobacterium longum), SzGlmU (Streptococcus zooepidemicus), PmPpA, and PmHAS 1À 703 (both from Pasteurella multocida).…”

Section: Introductionmentioning

confidence: 99%

“…Common industrial processes like extraction from rooster combs or fermentation with Streptococcus end up into a highly dispersed HA product, which also has the risk of contamination of avian proteins or endotoxins, respectively [6–9] . Therefore, enzymatic strategies to produce HA are the focus of research [10,11] . In our previous study, we established an in vitro one‐pot synthesis to produce high molecular weight (HMW) HA [12] .…”

Section: Introductionmentioning

confidence: 99%

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Integration of a Nucleoside Triphosphate Regeneration System in the One‐pot Synthesis of UDP‐sugars and Hyaluronic Acid

et al. 2021

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Enzymatic synthesis is a promising approach to produce hyaluronic acid (HA) products with defined molecular weight (MW) and dispersity. We here report on the evaluation of nucleoside triphosphate (NTP) regeneration system for the synthesis of the precursors uridine-5'-diphosphate α-d-N-acetylglucosamine (UDP-GlcNAc) and uridine-5'-diphosphate α-dglucuronic acid (UDP-GlcA) and integration in the one-pot synthesis of HA. By integration of polyphosphate kinase from Ruegeria pomeroyi (RpPPK2-3) into the enzyme cascade to synthesize UDP-GlcA we reached 89 % yield after 1 h with a regeneration cycle number (RCN) of 17. With the UDP-GlcNAc cascade, the ATP concentration was lowered 250-fold and reached 94 % yield after 1 h with an RCN of 234. Integration of the RpPPK2-3 regeneration system in one-pot HA synthesis afforded optimization of MgCl 2 and ATP starting concentrations. With 25 mM MgCl 2 and 0.1 mM ATP, a HA concentration of 0.81 g L À 1 with an average MW of 1.17 MDa and dispersity of about 1.08 and an RCN of 75 was obtained. We conclude that integration of NTP regeneration with RpPPK2-3 expanded the enzymatic possibilities to produce HA with lower range MW.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Integration of a Nucleoside Triphosphate Regeneration System in the One‐pot Synthesis of UDP‐sugars and Hyaluronic Acid

et al. 2021

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“…Although genetic modifications helped to unveil the roles of GAGs to some extent, synergistic effects and compensation mechanisms between different proteoglycans and GAGs suggested a higher complexity of GAGs-mediated stem cell behaviors ( Corsi et al, 2002 ; Wadhwa et al, 2007 ; Bachvarova et al, 2020 ). To better decipher the specific sulfation pattern of proteoglycans, chemical, and enzymatic synthesis of GAGs ( DeAngelis et al, 2013 ; Gottschalk and Elling, 2021 ) with well-defined structures as well as characterization techniques such as nuclear magnetic resonance spectroscopy (NMR), surface plasmon resonance (SPR), and chromatography have been utilized to better understanding the binding interactions between protein ligands and GAGs ( Li et al, 2016 ; Pomin and Wang, 2018 ; Vallet et al, 2021 ). These chemoenzymatically synthesized GAGs have mainly been used for exogenous addition to cellular and animal models with the objective to determining how GAGs interact with growth factors in vitro and in vivo .…”

Section: Structural and Functional Characteristics Of Proteoglycans And Glycosaminoglycansmentioning

confidence: 99%

Proteoglycans and Glycosaminoglycans in Stem Cell Homeostasis and Bone Tissue Regeneration

Chen

Sun

You

et al. 2021

Front. Cell Dev. Biol.

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Stem cells maintain a subtle balance between self-renewal and differentiation under the regulatory network supported by both intracellular and extracellular components. Proteoglycans are large glycoproteins present abundantly on the cell surface and in the extracellular matrix where they play pivotal roles in facilitating signaling transduction and maintaining stem cell homeostasis. In this review, we outline distinct proteoglycans profiles and their functions in the regulation of stem cell homeostasis, as well as recent progress and prospects of utilizing proteoglycans/glycosaminoglycans as a novel glycomics carrier or bio-active molecules in bone regeneration.

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“…HA is manufactured either by harsh extraction from rooster combs or fermentation with Streptococcus resulting in a highly disperse product and the risk of avian protein or endotoxin contamination [4,9–11] . Therefore, enzymatically oriented approaches to produce HA or other GAGs stay in focus of today‘s research [1] …”

Section: Introductionmentioning

confidence: 99%

“…[4,[9][10][11] Therefore, enzymatically oriented approaches to produce HA or other GAGs stay in focus of today's research. [1] Enzymatic catalysis has been intensively studied as a cleaner and green alternative and is increasingly important for the industry. [12][13][14][15] However, production costs, the need to improve stability, activity, and purity of enzymes are still obstacles for the industrial breakthrough.…”

Section: Introductionmentioning

confidence: 99%

Repetitive Synthesis of High‐Molecular‐Weight Hyaluronic Acid with Immobilized Enzyme Cascades

et al. 2021

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Industrial hyaluronic acid (HA) production comprises either fermentation with Streptococcus strains or extraction from rooster combs. The hard‐to‐control product quality is an obstacle to these processes. Enzymatic syntheses of HA were developed to produce high‐molecular‐weight HA with low dispersity. To facilitate enzyme recovery and biocatalyst re‐use, here the immobilization of cascade enzymes onto magnetic beads was used for the synthesis of uridine‐5’‐diphosphate‐α‐d‐N‐acetyl‐glucosamine (UDP‐GlcNAc), UDP‐glucuronic acid (UDP‐GlcA), and HA. The combination of six enzymes in the UDP‐sugar cascades with integrated adenosine‐5'‐triphosphate‐regeneration reached yields between 60 and 100 % for 5 repetitive batches, proving the productivity. Immobilized HA synthase from Pasteurella multocida produced HA in repetitive batches for three days. Combining all seven immobilized enzymes in a one‐pot synthesis, HA production was demonstrated for three days with a HA concentration of up to 0.37 g L−1, an average MW of 2.7–3.6 MDa, and a dispersity of 1.02–1.03.

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Current state on the enzymatic synthesis of glycosaminoglycans

Cited by 33 publications

References 69 publications

Integration of a Nucleoside Triphosphate Regeneration System in the One‐pot Synthesis of UDP‐sugars and Hyaluronic Acid

Integration of a Nucleoside Triphosphate Regeneration System in the One‐pot Synthesis of UDP‐sugars and Hyaluronic Acid

Proteoglycans and Glycosaminoglycans in Stem Cell Homeostasis and Bone Tissue Regeneration

Repetitive Synthesis of High‐Molecular‐Weight Hyaluronic Acid with Immobilized Enzyme Cascades

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