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Characterisation of two novel cyclodextrinases using on-line microdialysis sampling with high-performance anion exchange chromatography

Nilsson, Carina LU ; Nilsson, Frida ; Turner, Pernilla LU ; Sixtensson, Martin ; Nordberg Karlsson, Eva LU orcid ; Holst, Olle LU ; Cohen, Arieh LU and Gorton, Lo LU (2006) In Analytical and Bioanalytical Chemistry 385(8). p.1421-1429
Abstract
In this work, a real-time sampling/analytical method for on-line measurements of two newly discovered cyclomaltodextrinases (CDases) has been developed and evaluated. This novel methodology not only allows the final products to be investigated, but it also reveals enzyme-specific differences in the degradation pathways during the hydrolysis of different substrates, which is a great advantage in the important tasks of investigating the mechanisms of and classifying new hydrolases, and is an advantage that conventional techniques cannot offer. Two different enzymes, one CDase from Laceyella sacchari (LsCda13) and one from Anoxybacillus flavithermus (AfCda13), were investigated during the hydrolysis of alpha-, beta- and gamma-cyclodextrin,... (More)
In this work, a real-time sampling/analytical method for on-line measurements of two newly discovered cyclomaltodextrinases (CDases) has been developed and evaluated. This novel methodology not only allows the final products to be investigated, but it also reveals enzyme-specific differences in the degradation pathways during the hydrolysis of different substrates, which is a great advantage in the important tasks of investigating the mechanisms of and classifying new hydrolases, and is an advantage that conventional techniques cannot offer. Two different enzymes, one CDase from Laceyella sacchari (LsCda13) and one from Anoxybacillus flavithermus (AfCda13), were investigated during the hydrolysis of alpha-, beta- and gamma-cyclodextrin, and the hydrolysis products were sampled via a microdialysis probe and injected on-line every 30 min into a high-performance anion exchange chromatography system equipped with a pulsed amperometric detector (HPAEC-PAD), where they were identified. The enzymes yielded the same end-products, maltose and glucose, in an approximate molar ratio of 2:1, but they exhibited distinctly different patterns of intermediate product formation before reaching the end-point. LsCda13 had a more random distribution of the intermediate products, whereas AfCda13 showed the distinct intermediate production of maltotriose, which in some cases accumulated. (Less)
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Laceyella sacchari (LsCda13), enzyme hydrolysis, cyclodextrins (CD), carbohydrate analysis, Anoxybacillus flavithermus (AfCda13)
in
Analytical and Bioanalytical Chemistry
volume
385
issue
8
pages
1421 - 1429
publisher
Springer
external identifiers
  • wos:000239644200012
  • scopus:33746895433
  • pmid:16804672
ISSN
1618-2642
DOI
10.1007/s00216-006-0570-7
language
English
LU publication?
yes
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004), Biotechnology (LTH) (011001037), Division of Occupational and Environmental Medicine (013078001)
id
13bbea43-4740-468a-8983-febbd0063986 (old id 398564)
date added to LUP
2016-04-01 11:45:44
date last changed
2021-09-22 02:22:33
@article{13bbea43-4740-468a-8983-febbd0063986,
  abstract     = {In this work, a real-time sampling/analytical method for on-line measurements of two newly discovered cyclomaltodextrinases (CDases) has been developed and evaluated. This novel methodology not only allows the final products to be investigated, but it also reveals enzyme-specific differences in the degradation pathways during the hydrolysis of different substrates, which is a great advantage in the important tasks of investigating the mechanisms of and classifying new hydrolases, and is an advantage that conventional techniques cannot offer. Two different enzymes, one CDase from Laceyella sacchari (LsCda13) and one from Anoxybacillus flavithermus (AfCda13), were investigated during the hydrolysis of alpha-, beta- and gamma-cyclodextrin, and the hydrolysis products were sampled via a microdialysis probe and injected on-line every 30 min into a high-performance anion exchange chromatography system equipped with a pulsed amperometric detector (HPAEC-PAD), where they were identified. The enzymes yielded the same end-products, maltose and glucose, in an approximate molar ratio of 2:1, but they exhibited distinctly different patterns of intermediate product formation before reaching the end-point. LsCda13 had a more random distribution of the intermediate products, whereas AfCda13 showed the distinct intermediate production of maltotriose, which in some cases accumulated.},
  author       = {Nilsson, Carina and Nilsson, Frida and Turner, Pernilla and Sixtensson, Martin and Nordberg Karlsson, Eva and Holst, Olle and Cohen, Arieh and Gorton, Lo},
  issn         = {1618-2642},
  language     = {eng},
  number       = {8},
  pages        = {1421--1429},
  publisher    = {Springer},
  series       = {Analytical and Bioanalytical Chemistry},
  title        = {Characterisation of two novel cyclodextrinases using on-line microdialysis sampling with high-performance anion exchange chromatography},
  url          = {http://dx.doi.org/10.1007/s00216-006-0570-7},
  doi          = {10.1007/s00216-006-0570-7},
  volume       = {385},
  year         = {2006},
}