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Cloning and characterisation of Aspergillus niger genes encoding an alpha-galactosidase and a beta-mannosidase involved in galactomannan degradation

Ademark, Pia; de Vries, R P; Hägglund, Per LU ; Stålbrand, Henrik LU and Visser, J (2001) In European Journal of Biochemistry 268(10). p.2982-2990
Abstract
α-Galactosidase (EC 3.2.1.22) and β-mannosidase (EC 3.2.1.25) participate in the hydrolysis of complex plant saccharides such as galacto(gluco)mannans. Here we report on the cloning and characterization of genes encoding an α-galactosidase (AglC) and a β-mannosidase (MndA) from Aspergillus niger. The aglC and mndA genes code for 747 and 931 amino acids, respectively, including the eukaryotic signal sequences. The predicted isoelectric points of AglC and MndA are 4.56 and 5.17, and the calculated molecular masses are 79.674 and 102.335 kDa, respectively. Both AglC and MndA contain several putative N-glycosylation sites. AglC was assigned to family 36 of the glycosyl hydrolases and MndA was assigned to family 2. The expression patterns of... (More)
α-Galactosidase (EC 3.2.1.22) and β-mannosidase (EC 3.2.1.25) participate in the hydrolysis of complex plant saccharides such as galacto(gluco)mannans. Here we report on the cloning and characterization of genes encoding an α-galactosidase (AglC) and a β-mannosidase (MndA) from Aspergillus niger. The aglC and mndA genes code for 747 and 931 amino acids, respectively, including the eukaryotic signal sequences. The predicted isoelectric points of AglC and MndA are 4.56 and 5.17, and the calculated molecular masses are 79.674 and 102.335 kDa, respectively. Both AglC and MndA contain several putative N-glycosylation sites. AglC was assigned to family 36 of the glycosyl hydrolases and MndA was assigned to family 2. The expression patterns of aglC and mndA and two other genes encoding A. nigerα-galactosidases (aglA and aglB) during cultivation on galactomannan were studied by Northern analysis. A comparison of gene expression on monosaccharides in the A. niger wild-type and a CreA mutant strain showed that the carbon catabolite repressor protein CreA has a strong influence on aglA, but not on aglB, aglC or mndA. AglC and MndA were purified from constructed overexpression strains of A. niger, and the combined action of these enzymes degraded a galactomanno-oligosaccharide into galactose and mannose. The possible roles of AglC and MndA in galactomannan hydrolysis is discussed. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
α-galactosidase, expression., β-mannosidase, cloning, Aspergillus niger
in
European Journal of Biochemistry
volume
268
issue
10
pages
2982 - 2990
publisher
Wiley-Blackwell
external identifiers
  • scopus:0034819282
ISSN
0014-2956
DOI
language
English
LU publication?
yes
id
74c58119-0385-44e6-9626-28e3b0076a47 (old id 42307)
date added to LUP
2007-07-17 14:25:53
date last changed
2018-05-29 11:13:44
@article{74c58119-0385-44e6-9626-28e3b0076a47,
  abstract     = {α-Galactosidase (EC 3.2.1.22) and β-mannosidase (EC 3.2.1.25) participate in the hydrolysis of complex plant saccharides such as galacto(gluco)mannans. Here we report on the cloning and characterization of genes encoding an α-galactosidase (AglC) and a β-mannosidase (MndA) from Aspergillus niger. The aglC and mndA genes code for 747 and 931 amino acids, respectively, including the eukaryotic signal sequences. The predicted isoelectric points of AglC and MndA are 4.56 and 5.17, and the calculated molecular masses are 79.674 and 102.335 kDa, respectively. Both AglC and MndA contain several putative N-glycosylation sites. AglC was assigned to family 36 of the glycosyl hydrolases and MndA was assigned to family 2. The expression patterns of aglC and mndA and two other genes encoding A. nigerα-galactosidases (aglA and aglB) during cultivation on galactomannan were studied by Northern analysis. A comparison of gene expression on monosaccharides in the A. niger wild-type and a CreA mutant strain showed that the carbon catabolite repressor protein CreA has a strong influence on aglA, but not on aglB, aglC or mndA. AglC and MndA were purified from constructed overexpression strains of A. niger, and the combined action of these enzymes degraded a galactomanno-oligosaccharide into galactose and mannose. The possible roles of AglC and MndA in galactomannan hydrolysis is discussed.},
  author       = {Ademark, Pia and de Vries, R P and Hägglund, Per and Stålbrand, Henrik and Visser, J},
  issn         = {0014-2956},
  keyword      = {α-galactosidase,expression.,β-mannosidase,cloning,Aspergillus niger},
  language     = {eng},
  number       = {10},
  pages        = {2982--2990},
  publisher    = {Wiley-Blackwell},
  series       = {European Journal of Biochemistry},
  title        = {Cloning and characterisation of Aspergillus niger genes encoding an alpha-galactosidase and a beta-mannosidase involved in galactomannan degradation},
  url          = {http://dx.doi.org/},
  volume       = {268},
  year         = {2001},
}