The Discovery, Enzymatic Characterization and Functional Analysis of a Newly Isolated Chitinase from Marine-Derived Fungus <i>Aspergillus fumigatus</i> df347
In order to discover a broad-specificity and high stability chitinase, a marine fungus, <i>Aspergillus fumigatus</i> df347, was identified in the sediments of mangrove wetlands in Qinzhou Bay, China. The chitinase gene (<i>Af</i>Chi28) from <i>A. fumigatus</i> df3...
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| Format: | Article |
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MDPI AG
2022-08-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/20/8/520 |
| _version_ | 1797431970176696320 |
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| author | Ya-Li Wu Sheng Wang Deng-Feng Yang Li-Yan Yang Qing-Yan Wang Jun Yu Nan Li Li-Xia Pan |
| author_facet | Ya-Li Wu Sheng Wang Deng-Feng Yang Li-Yan Yang Qing-Yan Wang Jun Yu Nan Li Li-Xia Pan |
| author_sort | Ya-Li Wu |
| collection | DOAJ |
| description | In order to discover a broad-specificity and high stability chitinase, a marine fungus, <i>Aspergillus fumigatus</i> df347, was identified in the sediments of mangrove wetlands in Qinzhou Bay, China. The chitinase gene (<i>Af</i>Chi28) from <i>A. fumigatus</i> df347 was cloned and heterologously expressed in <i>Escherichia coli</i>, and the recombinant enzyme <i>Af</i>Chi28 was purified and characterized. <i>Af</i>Chi28 is an acido-halotolerant- and temperature-resistant bifunctional enzyme with both endo- and exo-cleavage functions. Its enzymatic products are mainly GlcNAc, (GlcNAc)<sub>2</sub>, (GlcNAc)<sub>3</sub> and (GlcNAc)<sub>4</sub>. Na<sup>+</sup>, Mg<sup>2+</sup>, K<sup>+</sup>, Ca<sup>2+</sup> and Tris at a concentration of 50 mM had a strong stimulatory effect on <i>Af</i>Chi28. The crude enzyme and pure enzyme exhibited the highest specific activity of 0.737 mU/mg and 52.414 mU/mg towards colloidal chitin. The DxDxE motif at the end of strand <i>β</i>5 and with Glu154 as the catalytic residue was verified by the AlphaFold2 prediction and sequence alignment of homologous proteins. Moreover, the results of molecular docking showed that molecular modeling of chitohexaose was shown to bind to <i>Af</i>Chi28 in subsites −4 to +2 in the deep groove substrate-binding pocket. This study demonstrates that <i>Af</i>Chi28 is a promising chitinase for the preparation of desirable chitin oligosaccharides, and provides a foundation for elucidating the catalytic mechanism of chitinases from marine fungi. |
| first_indexed | 2024-03-09T09:54:32Z |
| format | Article |
| id | doaj.art-953ff137f12c43498418153e15dd37f6 |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-09T09:54:32Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-953ff137f12c43498418153e15dd37f62023-12-01T23:56:06ZengMDPI AGMarine Drugs1660-33972022-08-0120852010.3390/md20080520The Discovery, Enzymatic Characterization and Functional Analysis of a Newly Isolated Chitinase from Marine-Derived Fungus <i>Aspergillus fumigatus</i> df347Ya-Li Wu0Sheng Wang1Deng-Feng Yang2Li-Yan Yang3Qing-Yan Wang4Jun Yu5Nan Li6Li-Xia Pan7College of Life Science and Technology, Guangxi University, Nanning 530004, ChinaNanning Pangbo Biological Engineering Co., Ltd., Nanning 530004, ChinaState Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Academy of Sciences, 98 Daling Road, Nanning 530007, ChinaState Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Academy of Sciences, 98 Daling Road, Nanning 530007, ChinaState Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Academy of Sciences, 98 Daling Road, Nanning 530007, ChinaState Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Academy of Sciences, 98 Daling Road, Nanning 530007, ChinaCollege of Life Science and Technology, Guangxi University, Nanning 530004, ChinaState Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Academy of Sciences, 98 Daling Road, Nanning 530007, ChinaIn order to discover a broad-specificity and high stability chitinase, a marine fungus, <i>Aspergillus fumigatus</i> df347, was identified in the sediments of mangrove wetlands in Qinzhou Bay, China. The chitinase gene (<i>Af</i>Chi28) from <i>A. fumigatus</i> df347 was cloned and heterologously expressed in <i>Escherichia coli</i>, and the recombinant enzyme <i>Af</i>Chi28 was purified and characterized. <i>Af</i>Chi28 is an acido-halotolerant- and temperature-resistant bifunctional enzyme with both endo- and exo-cleavage functions. Its enzymatic products are mainly GlcNAc, (GlcNAc)<sub>2</sub>, (GlcNAc)<sub>3</sub> and (GlcNAc)<sub>4</sub>. Na<sup>+</sup>, Mg<sup>2+</sup>, K<sup>+</sup>, Ca<sup>2+</sup> and Tris at a concentration of 50 mM had a strong stimulatory effect on <i>Af</i>Chi28. The crude enzyme and pure enzyme exhibited the highest specific activity of 0.737 mU/mg and 52.414 mU/mg towards colloidal chitin. The DxDxE motif at the end of strand <i>β</i>5 and with Glu154 as the catalytic residue was verified by the AlphaFold2 prediction and sequence alignment of homologous proteins. Moreover, the results of molecular docking showed that molecular modeling of chitohexaose was shown to bind to <i>Af</i>Chi28 in subsites −4 to +2 in the deep groove substrate-binding pocket. This study demonstrates that <i>Af</i>Chi28 is a promising chitinase for the preparation of desirable chitin oligosaccharides, and provides a foundation for elucidating the catalytic mechanism of chitinases from marine fungi.https://www.mdpi.com/1660-3397/20/8/520chitinasestrain screeningenzymatic propertieschitin oligosaccharidesheterologous expressionmolecular docking |
| spellingShingle | Ya-Li Wu Sheng Wang Deng-Feng Yang Li-Yan Yang Qing-Yan Wang Jun Yu Nan Li Li-Xia Pan The Discovery, Enzymatic Characterization and Functional Analysis of a Newly Isolated Chitinase from Marine-Derived Fungus <i>Aspergillus fumigatus</i> df347 Marine Drugs chitinase strain screening enzymatic properties chitin oligosaccharides heterologous expression molecular docking |
| title | The Discovery, Enzymatic Characterization and Functional Analysis of a Newly Isolated Chitinase from Marine-Derived Fungus <i>Aspergillus fumigatus</i> df347 |
| title_full | The Discovery, Enzymatic Characterization and Functional Analysis of a Newly Isolated Chitinase from Marine-Derived Fungus <i>Aspergillus fumigatus</i> df347 |
| title_fullStr | The Discovery, Enzymatic Characterization and Functional Analysis of a Newly Isolated Chitinase from Marine-Derived Fungus <i>Aspergillus fumigatus</i> df347 |
| title_full_unstemmed | The Discovery, Enzymatic Characterization and Functional Analysis of a Newly Isolated Chitinase from Marine-Derived Fungus <i>Aspergillus fumigatus</i> df347 |
| title_short | The Discovery, Enzymatic Characterization and Functional Analysis of a Newly Isolated Chitinase from Marine-Derived Fungus <i>Aspergillus fumigatus</i> df347 |
| title_sort | discovery enzymatic characterization and functional analysis of a newly isolated chitinase from marine derived fungus i aspergillus fumigatus i df347 |
| topic | chitinase strain screening enzymatic properties chitin oligosaccharides heterologous expression molecular docking |
| url | https://www.mdpi.com/1660-3397/20/8/520 |
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