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Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production

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’ lig...

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Autores principales: Manfrão-Netto, João Heitor Colombelli, Queiroz, Enzo Bento, Rodrigues, Kelly Assis, Coelho, Cintia M., Paes, Hugo Costa, Rech, Elibio Leopoldo, Parachin, Nádia Skorupa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7913781/
https://www.ncbi.nlm.nih.gov/pubmed/33546444
http://dx.doi.org/10.3390/microorganisms9020312
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author Manfrão-Netto, João Heitor Colombelli
Queiroz, Enzo Bento
Rodrigues, Kelly Assis
Coelho, Cintia M.
Paes, Hugo Costa
Rech, Elibio Leopoldo
Parachin, Nádia Skorupa
author_facet Manfrão-Netto, João Heitor Colombelli
Queiroz, Enzo Bento
Rodrigues, Kelly Assis
Coelho, Cintia M.
Paes, Hugo Costa
Rech, Elibio Leopoldo
Parachin, Nádia Skorupa
author_sort Manfrão-Netto, João Heitor Colombelli
collection PubMed
description 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.
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spelling pubmed-79137812021-02-28 Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production Manfrão-Netto, João Heitor Colombelli Queiroz, Enzo Bento Rodrigues, Kelly Assis Coelho, Cintia M. Paes, Hugo Costa Rech, Elibio Leopoldo Parachin, Nádia Skorupa Microorganisms Article 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. MDPI 2021-02-03 /pmc/articles/PMC7913781/ /pubmed/33546444 http://dx.doi.org/10.3390/microorganisms9020312 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Manfrão-Netto, João Heitor Colombelli
Queiroz, Enzo Bento
Rodrigues, Kelly Assis
Coelho, Cintia M.
Paes, Hugo Costa
Rech, Elibio Leopoldo
Parachin, Nádia Skorupa
Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production
title Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production
title_full Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production
title_fullStr Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production
title_full_unstemmed Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production
title_short Evaluation of Ogataea (Hansenula) polymorpha for Hyaluronic Acid Production
title_sort evaluation of ogataea (hansenula) polymorpha for hyaluronic acid production
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7913781/
https://www.ncbi.nlm.nih.gov/pubmed/33546444
http://dx.doi.org/10.3390/microorganisms9020312
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