Title
Exo-inulinase of Aspergillus niger N402: A hydrolytic enzyme with significant transfructosylating activity
Author
Goosen, C.
van der Maarel, M.J.E.C.
Dijkhuizen, L.
TNO Kwaliteit van Leven
Publication year
2008
Abstract
The purified exo-inulinase enzyme of Aspergillus niger N402 (AngInuE; heterologously expressed in Escherichia coli) displayed a sucrose:inulin (S/I) hydrolysis ratio of 2.3, characteristic for a typical exo-inulinase. The enzyme also had significant transfructosylating activity with increasing sucrose concentrations, producing various oligosaccharides. The AngInuE protein molecular mass was 57 kDa, close to the calculated value for the mature protein. AngInuE thus was active as a monomeric, non-glycosylated protein. Contradictory data on hydrolysis/transfructosylation activity ratios have been published for the (almost) identical (but monomeric or dimeric and glycosylated) exo-inulinases of other aspergilli. Our data clearly show that the AngInuE enzyme, produced in and purified from E. coli, is a broad specificity exo-inulinase that also has significant transfructosylating activity with sucrose. Analysis of site-directed mutants of AngInuE showed that the glycoside hydrolase family 32 conserved domain G is important for catalytic efficiency, with a clear role in hydrolysis of both sucrose and fructans.
Subject
Biology
Aspergillus niger
Exo-inulinase
Glycoside hydrolase
Hydrolysis
SVEVF motif
Transfructosylation
Enzymes
Escherichia coli
Hydrolysis
Polysaccharides
Purification
Sugar (sucrose)
Sugars
Water pollution
Aspergillus niger
Exo-inulinase
Glycoside hydrolase
SVEVF motif
Transfructosylation
Enzyme activity
fructan
fungal enzyme
glycosidase
hydrolase
inulinase
oligosaccharide
protein AngInuE
sucrose
article
Aspergillus niger
carbohydrate synthesis
catalysis
concentration response
enzyme activity
enzyme analysis
enzyme purification
enzyme specificity
enzyme substrate
enzyme synthesis
Escherichia coli
fungal strain
hydrolysis
molecular weight
nonhuman
nucleotide sequence
protein analysis
protein domain
site directed mutagenesis
Aspergillus niger
Escherichia coli
To reference this document use:
http://resolver.tudelft.nl/uuid:2a150c73-272e-4085-8528-c4f12450e3d3
DOI
https://doi.org/10.1080/10242420701806686
TNO identifier
241000
ISSN
1024-2422
Source
Biocatalysis and Biotransformation, 26 (1-2), 49-58
Document type
article