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...

Descripción completa

Detalles Bibliográficos
Autores principales: Liu, Hongfei, Ren, Wei, Ly, Mingsheng, Li, Haifeng, Wang, Shujun
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