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 ( Ca Aga1) from C. algicola was heterologously expressed in Escherichia coli . The purified recombinant...
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ftdoajarticles:oai:doaj.org/article:77cefb7d7a4f4c6f813e257e1f7d1c1f 2023-05-15T13:43:41+02:00 Biochemical Characterization of a New β-Agarase from Cellulophaga algicola Zhenggang Han Yuxi Zhang Jiangke Yang 2019-04-01T00:00:00Z https://doi.org/10.3390/ijms20092143 https://doaj.org/article/77cefb7d7a4f4c6f813e257e1f7d1c1f EN eng MDPI AG https://www.mdpi.com/1422-0067/20/9/2143 https://doaj.org/toc/1422-0067 1422-0067 doi:10.3390/ijms20092143 https://doaj.org/article/77cefb7d7a4f4c6f813e257e1f7d1c1f International Journal of Molecular Sciences, Vol 20, Iss 9, p 2143 (2019) agar agarose β-agarase glycoside hydrolase neoagarooligosaccharide Biology (General) QH301-705.5 Chemistry QD1-999 article 2019 ftdoajarticles https://doi.org/10.3390/ijms20092143 2022-12-30T23:42:07Z 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 ( Ca Aga1) from C. algicola was heterologously expressed in Escherichia coli . The purified recombinant Ca Aga1 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. Ca Aga1 exhibited an optimal temperature and pH of 40 °C and 7, respectively. Ca Aga1 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 Ca Aga1 were 1.19 mg/mL and 36.21 U/mg, respectively. The reducing reagent (dithiothreitol) enhanced the activity of Ca Aga1 by more than one fold. In addition, Ca Aga1 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 Ca Aga1 is an endo-type β-agarase and efficiently hydrolyzed agarose into neoagarotetraose (NA4) and neoagarohexaose (NA6). A structural model of Ca Aga1 in complex with neoagarooctaose (NA8) was built by homology modeling and explained the hydrolysis pattern of Ca Aga1. Article in Journal/Newspaper Antarc* Antarctic Directory of Open Access Journals: DOAJ Articles Antarctic International Journal of Molecular Sciences 20 9 2143 |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
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language |
English |
topic |
agar agarose β-agarase glycoside hydrolase neoagarooligosaccharide Biology (General) QH301-705.5 Chemistry QD1-999 |
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agar agarose β-agarase glycoside hydrolase neoagarooligosaccharide Biology (General) QH301-705.5 Chemistry QD1-999 Zhenggang Han Yuxi Zhang Jiangke Yang Biochemical Characterization of a New β-Agarase from Cellulophaga algicola |
topic_facet |
agar agarose β-agarase glycoside hydrolase neoagarooligosaccharide Biology (General) QH301-705.5 Chemistry QD1-999 |
description |
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 ( Ca Aga1) from C. algicola was heterologously expressed in Escherichia coli . The purified recombinant Ca Aga1 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. Ca Aga1 exhibited an optimal temperature and pH of 40 °C and 7, respectively. Ca Aga1 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 Ca Aga1 were 1.19 mg/mL and 36.21 U/mg, respectively. The reducing reagent (dithiothreitol) enhanced the activity of Ca Aga1 by more than one fold. In addition, Ca Aga1 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 Ca Aga1 is an endo-type β-agarase and efficiently hydrolyzed agarose into neoagarotetraose (NA4) and neoagarohexaose (NA6). A structural model of Ca Aga1 in complex with neoagarooctaose (NA8) was built by homology modeling and explained the hydrolysis pattern of Ca Aga1. |
format |
Article in Journal/Newspaper |
author |
Zhenggang Han Yuxi Zhang Jiangke Yang |
author_facet |
Zhenggang Han Yuxi Zhang Jiangke Yang |
author_sort |
Zhenggang Han |
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 AG |
publishDate |
2019 |
url |
https://doi.org/10.3390/ijms20092143 https://doaj.org/article/77cefb7d7a4f4c6f813e257e1f7d1c1f |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
International Journal of Molecular Sciences, Vol 20, Iss 9, p 2143 (2019) |
op_relation |
https://www.mdpi.com/1422-0067/20/9/2143 https://doaj.org/toc/1422-0067 1422-0067 doi:10.3390/ijms20092143 https://doaj.org/article/77cefb7d7a4f4c6f813e257e1f7d1c1f |
op_doi |
https://doi.org/10.3390/ijms20092143 |
container_title |
International Journal of Molecular Sciences |
container_volume |
20 |
container_issue |
9 |
container_start_page |
2143 |
_version_ |
1766191838703648768 |