Biochemical Characterization of a New β-Agarase from Cellulophaga algicola
Cellulophaga algicola DSM 14237, isolated from the Eastern Antarctic coastal zone, was found to be able to hydrolyze several types of polysaccharide materials. In this study, a predicted β-agarase (CaAga1) from C. algicola was heterologously expressed in Escherichia coli. The purified recombinant Ca...
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ftpubmed:oai:pubmedcentral.nih.gov:6539560 2023-05-15T13:59:30+02:00 Biochemical Characterization of a New β-Agarase from Cellulophaga algicola Han, Zhenggang Zhang, Yuxi Yang, Jiangke 2019-04-30 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6539560/ http://www.ncbi.nlm.nih.gov/pubmed/31052274 https://doi.org/10.3390/ijms20092143 en eng MDPI http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6539560/ http://www.ncbi.nlm.nih.gov/pubmed/31052274 http://dx.doi.org/10.3390/ijms20092143 © 2019 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/). CC-BY Article Text 2019 ftpubmed https://doi.org/10.3390/ijms20092143 2019-06-09T00:22:17Z Cellulophaga algicola DSM 14237, isolated from the Eastern Antarctic coastal zone, was found to be able to hydrolyze several types of polysaccharide materials. In this study, a predicted β-agarase (CaAga1) from C. algicola was heterologously expressed in Escherichia coli. The purified recombinant CaAga1 showed specific activities of 29.39, 20.20, 14.12, and 8.99 U/mg toward agarose, pure agar, and crude agars from Gracilaria lemaneiformis and Porphyra haitanensis, respectively. CaAga1 exhibited an optimal temperature and pH of 40 °C and 7, respectively. CaAga1 was stable over a wide pH range from 4 to 11. The recombinant enzyme showed an unusual thermostability, that is, it was stable at temperature below or equal to 40 °C and around 70 °C, but was thermolabile at about 50 °C. With the agarose as the substrate, the K(m) and V(max) values for CaAga1 were 1.19 mg/mL and 36.21 U/mg, respectively. The reducing reagent (dithiothreitol) enhanced the activity of CaAga1 by more than one fold. In addition, CaAga1 was salt-tolerant given that it retained approximately 70% of the maximum activity in the presence of 2 M NaCl. The thin layer chromatography results indicated that CaAga1 is an endo-type β-agarase and efficiently hydrolyzed agarose into neoagarotetraose (NA4) and neoagarohexaose (NA6). A structural model of CaAga1 in complex with neoagarooctaose (NA8) was built by homology modeling and explained the hydrolysis pattern of CaAga1. Text Antarc* Antarctic PubMed Central (PMC) Antarctic International Journal of Molecular Sciences 20 9 2143 |
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Article Han, Zhenggang Zhang, Yuxi Yang, Jiangke Biochemical Characterization of a New β-Agarase from Cellulophaga algicola |
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Cellulophaga algicola DSM 14237, isolated from the Eastern Antarctic coastal zone, was found to be able to hydrolyze several types of polysaccharide materials. In this study, a predicted β-agarase (CaAga1) from C. algicola was heterologously expressed in Escherichia coli. The purified recombinant CaAga1 showed specific activities of 29.39, 20.20, 14.12, and 8.99 U/mg toward agarose, pure agar, and crude agars from Gracilaria lemaneiformis and Porphyra haitanensis, respectively. CaAga1 exhibited an optimal temperature and pH of 40 °C and 7, respectively. CaAga1 was stable over a wide pH range from 4 to 11. The recombinant enzyme showed an unusual thermostability, that is, it was stable at temperature below or equal to 40 °C and around 70 °C, but was thermolabile at about 50 °C. With the agarose as the substrate, the K(m) and V(max) values for CaAga1 were 1.19 mg/mL and 36.21 U/mg, respectively. The reducing reagent (dithiothreitol) enhanced the activity of CaAga1 by more than one fold. In addition, CaAga1 was salt-tolerant given that it retained approximately 70% of the maximum activity in the presence of 2 M NaCl. The thin layer chromatography results indicated that CaAga1 is an endo-type β-agarase and efficiently hydrolyzed agarose into neoagarotetraose (NA4) and neoagarohexaose (NA6). A structural model of CaAga1 in complex with neoagarooctaose (NA8) was built by homology modeling and explained the hydrolysis pattern of CaAga1. |
format |
Text |
author |
Han, Zhenggang Zhang, Yuxi Yang, Jiangke |
author_facet |
Han, Zhenggang Zhang, Yuxi Yang, Jiangke |
author_sort |
Han, Zhenggang |
title |
Biochemical Characterization of a New β-Agarase from Cellulophaga algicola |
title_short |
Biochemical Characterization of a New β-Agarase from Cellulophaga algicola |
title_full |
Biochemical Characterization of a New β-Agarase from Cellulophaga algicola |
title_fullStr |
Biochemical Characterization of a New β-Agarase from Cellulophaga algicola |
title_full_unstemmed |
Biochemical Characterization of a New β-Agarase from Cellulophaga algicola |
title_sort |
biochemical characterization of a new β-agarase from cellulophaga algicola |
publisher |
MDPI |
publishDate |
2019 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6539560/ http://www.ncbi.nlm.nih.gov/pubmed/31052274 https://doi.org/10.3390/ijms20092143 |
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Antarctic |
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Antarctic |
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Antarc* Antarctic |
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Antarc* Antarctic |
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6539560/ http://www.ncbi.nlm.nih.gov/pubmed/31052274 http://dx.doi.org/10.3390/ijms20092143 |
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© 2019 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/). |
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https://doi.org/10.3390/ijms20092143 |
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International Journal of Molecular Sciences |
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20 |
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