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Biochemical and Molecular Characteristics of a Novel Hyaluronic Acid Lyase from Citrobacter freundii
The Gram-negative strain of Citrobacter freundii, YNLX, has the ability to degrade hyaluronic acid. In this study, we expressed a C. freundii hyaluronic acid lyase, from polysaccharide lyase family 8, in Escherichia coli. The purified recombinant enzyme (rHynACF8) showed a substantially higher cleav...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9265501/ https://www.ncbi.nlm.nih.gov/pubmed/35804804 http://dx.doi.org/10.3390/foods11131989 |
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author | Li, Xinyue Li, Fang Ma, Junhao Li, Mingjun Lei, Xi Tang, Xianghua Wu, Qian Huang, Zunxi Zhang, Rui |
author_facet | Li, Xinyue Li, Fang Ma, Junhao Li, Mingjun Lei, Xi Tang, Xianghua Wu, Qian Huang, Zunxi Zhang, Rui |
author_sort | Li, Xinyue |
collection | PubMed |
description | The Gram-negative strain of Citrobacter freundii, YNLX, has the ability to degrade hyaluronic acid. In this study, we expressed a C. freundii hyaluronic acid lyase, from polysaccharide lyase family 8, in Escherichia coli. The purified recombinant enzyme (rHynACF8) showed a substantially higher cleavage activity of hyaluronic acid than chondroitin sulfate. We found that its optimal pH and temperature are 5.5 and 35 °C, respectively. In addition, the enzyme activity was not notably affected by most metal ions. K(m) and k(cat) of rHynACF8 towards HA were 1.5 ± 0.01 mg/mL and 30.9 ± 0.5 /s, respectively. rHynACF8 is an endo-acting enzyme. Its cleavage products had dramatically increased antioxidant activity than hyaluronic acid in vitro (p < 0.001). As the molecular weight of hyaluronic acid decreased, the intramolecular interactions among antioxidant functional groups were removed; in the process of the cracking reaction, new double bonds formed and conjugated with the carbonyl group. We presumed that the structural change is the critical factor influencing antioxidant capacity. Overall, we found that rHynACF8 from Gram-negative bacteria with metal ion resistance, indicated the relationship between the function and structure of its antioxidant cleavage product. |
format | Online Article Text |
id | pubmed-9265501 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-92655012022-07-09 Biochemical and Molecular Characteristics of a Novel Hyaluronic Acid Lyase from Citrobacter freundii Li, Xinyue Li, Fang Ma, Junhao Li, Mingjun Lei, Xi Tang, Xianghua Wu, Qian Huang, Zunxi Zhang, Rui Foods Article The Gram-negative strain of Citrobacter freundii, YNLX, has the ability to degrade hyaluronic acid. In this study, we expressed a C. freundii hyaluronic acid lyase, from polysaccharide lyase family 8, in Escherichia coli. The purified recombinant enzyme (rHynACF8) showed a substantially higher cleavage activity of hyaluronic acid than chondroitin sulfate. We found that its optimal pH and temperature are 5.5 and 35 °C, respectively. In addition, the enzyme activity was not notably affected by most metal ions. K(m) and k(cat) of rHynACF8 towards HA were 1.5 ± 0.01 mg/mL and 30.9 ± 0.5 /s, respectively. rHynACF8 is an endo-acting enzyme. Its cleavage products had dramatically increased antioxidant activity than hyaluronic acid in vitro (p < 0.001). As the molecular weight of hyaluronic acid decreased, the intramolecular interactions among antioxidant functional groups were removed; in the process of the cracking reaction, new double bonds formed and conjugated with the carbonyl group. We presumed that the structural change is the critical factor influencing antioxidant capacity. Overall, we found that rHynACF8 from Gram-negative bacteria with metal ion resistance, indicated the relationship between the function and structure of its antioxidant cleavage product. MDPI 2022-07-05 /pmc/articles/PMC9265501/ /pubmed/35804804 http://dx.doi.org/10.3390/foods11131989 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Li, Xinyue Li, Fang Ma, Junhao Li, Mingjun Lei, Xi Tang, Xianghua Wu, Qian Huang, Zunxi Zhang, Rui Biochemical and Molecular Characteristics of a Novel Hyaluronic Acid Lyase from Citrobacter freundii |
title | Biochemical and Molecular Characteristics of a Novel Hyaluronic Acid Lyase from Citrobacter freundii |
title_full | Biochemical and Molecular Characteristics of a Novel Hyaluronic Acid Lyase from Citrobacter freundii |
title_fullStr | Biochemical and Molecular Characteristics of a Novel Hyaluronic Acid Lyase from Citrobacter freundii |
title_full_unstemmed | Biochemical and Molecular Characteristics of a Novel Hyaluronic Acid Lyase from Citrobacter freundii |
title_short | Biochemical and Molecular Characteristics of a Novel Hyaluronic Acid Lyase from Citrobacter freundii |
title_sort | biochemical and molecular characteristics of a novel hyaluronic acid lyase from citrobacter freundii |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9265501/ https://www.ncbi.nlm.nih.gov/pubmed/35804804 http://dx.doi.org/10.3390/foods11131989 |
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