Cargando…
Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide
A GH49 dextranase gene DexKQ was cloned from marine bacteria Arthrobacter oxydans KQ11. It was recombinantly expressed using an Escherichia coli system. Recombinant DexKQ dextranase of 66 kDa exhibited the highest catalytic activity at pH 9.0 and 55 °C. kcat/Km of recombinant DexKQ at the optimum co...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6723167/ https://www.ncbi.nlm.nih.gov/pubmed/31430863 http://dx.doi.org/10.3390/md17080479 |
_version_ | 1783448706289238016 |
---|---|
author | Liu, Hongfei Ren, Wei Ly, Mingsheng Li, Haifeng Wang, Shujun |
author_facet | Liu, Hongfei Ren, Wei Ly, Mingsheng Li, Haifeng Wang, Shujun |
author_sort | Liu, Hongfei |
collection | PubMed |
description | A GH49 dextranase gene DexKQ was cloned from marine bacteria Arthrobacter oxydans KQ11. It was recombinantly expressed using an Escherichia coli system. Recombinant DexKQ dextranase of 66 kDa exhibited the highest catalytic activity at pH 9.0 and 55 °C. kcat/Km of recombinant DexKQ at the optimum condition reached 3.03 s(−1) μM(−1), which was six times that of commercial dextranase (0.5 s(−1) μM(−1)). DexKQ possessed a Km value of 67.99 µM against dextran T70 substrate with 70 kDa molecular weight. Thin-layer chromatography (TLC) analysis showed that main hydrolysis end products were isomalto-oligosaccharide (IMO) including isomaltotetraose, isomaltopantose, and isomaltohexaose. When compared with glucose, IMO could significantly improve growth of Bifidobacterium longum and Lactobacillus rhamnosus and inhibit growth of Escherichia coli and Staphylococcus aureus. This is the first report of dextranase from marine bacteria concerning recombinant expression and application in isomalto-oligosaccharide preparation. |
format | Online Article Text |
id | pubmed-6723167 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-67231672019-09-10 Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide Liu, Hongfei Ren, Wei Ly, Mingsheng Li, Haifeng Wang, Shujun Mar Drugs Article A GH49 dextranase gene DexKQ was cloned from marine bacteria Arthrobacter oxydans KQ11. It was recombinantly expressed using an Escherichia coli system. Recombinant DexKQ dextranase of 66 kDa exhibited the highest catalytic activity at pH 9.0 and 55 °C. kcat/Km of recombinant DexKQ at the optimum condition reached 3.03 s(−1) μM(−1), which was six times that of commercial dextranase (0.5 s(−1) μM(−1)). DexKQ possessed a Km value of 67.99 µM against dextran T70 substrate with 70 kDa molecular weight. Thin-layer chromatography (TLC) analysis showed that main hydrolysis end products were isomalto-oligosaccharide (IMO) including isomaltotetraose, isomaltopantose, and isomaltohexaose. When compared with glucose, IMO could significantly improve growth of Bifidobacterium longum and Lactobacillus rhamnosus and inhibit growth of Escherichia coli and Staphylococcus aureus. This is the first report of dextranase from marine bacteria concerning recombinant expression and application in isomalto-oligosaccharide preparation. MDPI 2019-08-19 /pmc/articles/PMC6723167/ /pubmed/31430863 http://dx.doi.org/10.3390/md17080479 Text en © 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/). |
spellingShingle | Article Liu, Hongfei Ren, Wei Ly, Mingsheng Li, Haifeng Wang, Shujun Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide |
title | Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter
oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide |
title_full | Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter
oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide |
title_fullStr | Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter
oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide |
title_full_unstemmed | Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter
oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide |
title_short | Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter
oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide |
title_sort | characterization of an alkaline gh49 dextranase from marine bacterium arthrobacter
oxydans kq11 and its application in the preparation of isomalto-oligosaccharide |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6723167/ https://www.ncbi.nlm.nih.gov/pubmed/31430863 http://dx.doi.org/10.3390/md17080479 |
work_keys_str_mv | AT liuhongfei characterizationofanalkalinegh49dextranasefrommarinebacteriumarthrobacteroxydanskq11anditsapplicationinthepreparationofisomaltooligosaccharide AT renwei characterizationofanalkalinegh49dextranasefrommarinebacteriumarthrobacteroxydanskq11anditsapplicationinthepreparationofisomaltooligosaccharide AT lymingsheng characterizationofanalkalinegh49dextranasefrommarinebacteriumarthrobacteroxydanskq11anditsapplicationinthepreparationofisomaltooligosaccharide AT lihaifeng characterizationofanalkalinegh49dextranasefrommarinebacteriumarthrobacteroxydanskq11anditsapplicationinthepreparationofisomaltooligosaccharide AT wangshujun characterizationofanalkalinegh49dextranasefrommarinebacteriumarthrobacteroxydanskq11anditsapplicationinthepreparationofisomaltooligosaccharide |